ACS Publications. Most Trusted. Most Cited. Most Read
MUC1 Glycopeptide Vaccine Modified with a GalNAc Glycocluster Targets the Macrophage Galactose C-type Lectin on Dendritic Cells to Elicit an Improved Humoral Response
More

OR SEARCH CITATIONS

My Activity
Publications

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:J. Am. Chem. Soc. 2023, 145, 24, 13027-13037
  • Open Access
Article

MUC1 Glycopeptide Vaccine Modified with a GalNAc Glycocluster Targets the Macrophage Galactose C-type Lectin on Dendritic Cells to Elicit an Improved Humoral Response
Click to copy article linkArticle link copied!

  • Adele Gabba*
    Adele Gabba
    School of Biological and Chemical Sciences, University of Galway, University Rd., H91 TK33 Galway, Ireland
    Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany
    *Email: [email protected]
    More by Adele Gabba
    Orcidhttps://orcid.org/0000-0001-8240-6482
  • Riem Attariya
    Riem Attariya
    Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    More by Riem Attariya
  • Sandra Behren
    Sandra Behren
    Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    More by Sandra Behren
  • Christian Pett
    Christian Pett
    Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    More by Christian Pett
  • Joost C. van der Horst
    Joost C. van der Horst
    Department of Molecular Cell Biology and Immunology, Amsterdam UMC Location Vrije Universiteit Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands
    Amsterdam Institute for Infection and Immunity, Cancer Immunology1105 AZ Amsterdam, the Netherlands
    More by Joost C. van der Horst
  • Hajime Yurugi
    Hajime Yurugi
    Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    More by Hajime Yurugi
  • Jin Yu
    Jin Yu
    Glycosciences Laboratory, Imperial College London, W12 0NN London, U.K.
    More by Jin Yu
  • Moritz Urschbach
    Moritz Urschbach
    Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany
    More by Moritz Urschbach
  • Juan Sabin
    Juan Sabin
    AFFINImeter Scientific & Development Team, Software 4 Science Developments, 15782 Santiago de Compostela, A Coruña, Spain
    Departamento de Física Aplicada, Facultad de Física, Universidad de Santiago de Compostela, 15705 Santiago de Compostela, Spain
    More by Juan Sabin
  • Gabriel Birrane
    Gabriel Birrane
    Division of Experimental Medicine, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, United States
    More by Gabriel Birrane
    Orcidhttps://orcid.org/0000-0002-1759-5499
  • Edgar Schmitt
    Edgar Schmitt
    Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    More by Edgar Schmitt
  • Sandra J. van Vliet
    Sandra J. van Vliet
    Department of Molecular Cell Biology and Immunology, Amsterdam UMC Location Vrije Universiteit Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands
    Amsterdam Institute for Infection and Immunity, Cancer Immunology1105 AZ Amsterdam, the Netherlands
    More by Sandra J. van Vliet
  • Pol Besenius
    Pol Besenius
    Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany
    More by Pol Besenius
    Orcidhttps://orcid.org/0000-0001-7478-4459
  • Ulrika Westerlind
    Ulrika Westerlind
    Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    More by Ulrika Westerlind
  • Paul V. Murphy
    Paul V. Murphy
    School of Biological and Chemical Sciences, University of Galway, University Rd., H91 TK33 Galway, Ireland
    SSPC, the Science Foundation Ireland Research Centre for Pharmaceuticals, University of Galway, University Rd., H91 TK33 Galway, Ireland
    More by Paul V. Murphy
    Orcidhttps://orcid.org/0000-0002-1529-6540
Open PDFSupporting Information (1)

Journal of the American Chemical Society

Cite this: J. Am. Chem. Soc. 2023, 145, 24, 13027–13037
Click to copy citationCitation copied!
https://doi.org/10.1021/jacs.2c12843
Published June 6, 2023

Copyright © 2023 The Authors. Published by American Chemical Society. This publication is licensed under

CC-BY 4.0 .

Abstract

Click to copy section linkSection link copied!
High Resolution Image
Download MS PowerPoint Slide

Mucin expression and glycosylation patterns on cancer cells differ markedly from healthy cells. Mucin 1 (MUC1) is overexpressed in several solid tumors and presents high levels of aberrant, truncated O-glycans (e.g., Tn antigen). Dendritic cells (DCs) express lectins that bind to these tumor-associated carbohydrate antigens (TACAs) to modulate immune responses. Selectively targeting these receptors with synthetic TACAs is a promising strategy to develop anticancer vaccines and to overcome TACA tolerance. In this work, we prepared, via a solid phase peptide synthesis approach, a modular tripartite vaccine candidate, incorporating a high-affinity glycocluster based on a tetraphenylethylene scaffold, to target the macrophage galactose-type lectin (MGL) on antigen presenting cells. MGL is a C-type lectin receptor that binds Tn antigens and can route them to human leukocyte antigen class II or I, making it an attractive target for anticancer vaccines. Conjugation of the glycocluster to a library of MUC1 glycopeptides bearing the Tn antigen is shown to promote uptake and recognition of the TACA by DCs via MGL. In vivo testing revealed that immunization with the newly designed vaccine construct bearing the GalNAc glycocluster induced a higher titer of anti-Tn-MUC1 antibodies compared to the TACAs alone. Additionally, the antibodies obtained bind a library of tumor-associated saccharide structures on MUC1 and MUC1-positive breast cancer cells. Conjugation of a high-affinity ligand for MGL to tumor-associated MUC1 glycopeptide antigens has a synergistic impact on antibody production.

This publication is licensed under

CC-BY 4.0 .
  • cc licence
  • by licence
Copyright © 2023 The Authors. Published by American Chemical Society

Subjects

what are subjects

Introduction

Click to copy section linkSection link copied!
The macrophage galactose-type lectin (MGL) is an endocytic receptor located on the surface of antigen-presenting cells (APCs), in particular dendritic cells (DCs) and macrophages. MGL binds to terminal α- or β-GalNAc residues in a calcium-dependent manner (1−3) and can employ a secondary binding site outside the carbohydrate recognition domain (CRD) to engage with glycoproteins (4) or disaccharides. (5) MGL was reported to recognize Tn-modified Mucin 1 (Tn-MUC1) tandem repeat (TR) glycopeptides (6,7) on several cancer cell lines, including colon, lung, and bronchioalveolar carcinoma. (8) The MUC1 glycoprotein is a membrane-bound mucin found on epithelial cells, but aberrantly expressed with truncated glycans in several cancer cell lines. (9−12)
These unique MUC1 glycopeptide epitopes on epithelial cancer cells are interesting targets for immunotherapy. (13,14) However, MUC1 immunotolerance is a major hurdle in the development of efficient anticancer vaccines. (15) The unglycosylated protein backbone is not sufficient to trigger an immune response and unconjugated MUC1 glycopeptides are weakly immunogenic.
The role of MGL in immune regulation is not fully understood but appears to be highly dependent on the antigen structure. In some cases, glycotope interactions with MGL seem to facilitate immune escape of the pathogen via DC secretion of anti-inflammatory cytokines, (16−19) such as interleukin-10 (IL-10), (20) which reduces the glycolytic activity of DCs (5) or abrogates the T helper 1/2 (Th1/2) type response in favor of regulatory T cells (Treg) or T helper 17 (Th17) type responses. (21)
However, in other cases, the Tn–MGL interaction has been leveraged to prime the immune system against tumor antigens by promoting antigen processing and presentation on APCs.
A glycopeptide therapeutic vaccine candidate based on multiple antigenic glycopeptide (MAG) composed of tri Tn glycotope, MAG-Tn3, has been shown to engage MGL to induce tumor-specific cytotoxic antibodies in breast cancer patients. (22−24) In another study, a short peptide lacking glycosylation was bound to the MGL CRD in a calcium-dependent manner and was shown to improve survival in an ovarian cancer murine model by triggering IFN-γ release and maturation of immune cells. (25) Furthermore, MGL expressed specifically on CD1c+ DCs was reported to bind and internalize Tn-MUC1 glycopeptides and enhance toll-like receptor 7/8-induced cytokine secretion. (26)
The dichotomous signaling and consequent immune response observed upon MGL engagement requires further investigation, but the remarkable example offered by Leclerc team with MAG-Tn3, and the solid evidence that MGL enhances antigen uptake and routing of tumor-associated carbohydrate antigens (TACA’s) to human leukocyte antigen class I and II compartments (27,28) suggest that MGL targeting for prophylactic and therapeutic vaccine development should not be disregarded.
Herein, we propose to further enhance MGL-induced Tn-MUC1 uptake by DCs via a modular vaccine design strategy and incorporate a synthetic glycocluster into vaccine constructs with an aim to overcome the natural immunotolerance of MUC1.
We report the synthesis of a trivalent ligand 1, which can be used in Fmoc solid-phase peptide synthesis (Fmoc-SPPS). This synthetic glycocluster has three α-S-GalNAc residues grafted to a tetraphenylethylene (TPE) scaffold containing an amino acid linker (TPEaa, 1). Specifically, we generated a glycopeptide library based on the MUC1 TR HGVTSAPDT*RPAPGS*T*AP (* represents a possible glycosylation site) presenting the Tn-antigen at the possible glycosylation sites and the TPEaa 1 at the peptide’s N-terminus. We assessed conjugate binding to recombinant MGL (2) via enzyme linked immunosorbent assay (ELISA) and found that the TPEaa-functionalized MUC1 enhanced binding to MGL. The presence of the TPEaa glycocluster 1 increased the peptide avidity for MGL approximately 10-fold (11 vs 12 and 13). For in vivo studies, the novel fully synthetic tripartite vaccine candidate 16, consisting of the trivalent TPE glycocluster 1, the P30 T helper epitope, and an antigenic Tn-modified MUC1 glycopeptide was synthesized. The tripartite construct 16, induced a higher antibody titer compared to the control 15 that lacked the glycocluster unit 1. The antibodies produced upon immunization with 16 bound to MUC1-Tn-positive T47D human breast cancer cells. Furthermore, the antibody selectivity was assessed via glycopeptide microarray binding assays, which revealed that the presence of glycocluster 1 did not alter the recognition pattern of the antibodies.
Both sera showed a preferential binding for the PDTR region over glycopeptides glycosylated at the GVTS or GSTA regions while binding for extensively glycosylated glycopeptides, such as core 2 and core 4 common for healthy cells mucins, was not observed.

Results and Discussion

Click to copy section linkSection link copied!

Glycan and Multivalent Ligand Design

MGL CRD preferentially binds GalNAc over galactopyranoside mostly due to direct hydrogen bond between the carbonyl of the acetamide group and Nε2 of the His286 imidazole group and CH−π interaction between the methyl group of the acetamide with the aromatic side chain of Tyr236. (29) It shows a slight preference for α-Gal over the corresponding β-anomer. (2,30) Furthermore, monovalent α-S-GalNAc had a 50-fold higher inhibitory potency than GalNAc for MGL CRD bound to crypt-associated cells of murine jejunum. (31) Thioglycosides (S-glycosides) are accepted by most biological systems and are less susceptible to acid or enzymatic cleavage than O-glycosides. Given its enhanced affinity and well-established synthetic access, we designed a multivalent glycocluster display of the α-S-GalNAc derivative I (Scheme 1).

Scheme 1

Scheme 1. Synthesis of the Trivalent Ligand 1 to Engage MGL
High Resolution Image
Download MS PowerPoint Slide
We previously assessed the ability of tetravalent and trivalent α-S-GalNAc glycoclusters in inhibiting the binding of MGL to fixed murine jejunum. The ligands containing the TPE scaffold outperformed other scaffolds with the same valency and similar geometry. (31) This might be due to the established ability of TPE to aggregate in aqueous media (32,33) with the increased valency of the supramolecular structure leading to higher-affinity interactions with MGL. Before embarking on the design of a synthetic route for a conjugatable glycocluster based on the TPE core, we evaluated the ability of the glycocluster 1S (Figure S2) to bind human MGL (hMGL) by isothermal titration calorimetry (ITC). We expressed recombinantly and purified the hMGL CRD, amino acids 181–277. To mimic closely the oligomerization state of the protein on APCs, also the extracellular portion of the lectin, amino acids 81–309, containing a coiled-coil motive, amino acids 85–176, (34) that induces the trimerization of the CRD was recombinantly expressed and purified (Figure S1).
ITC experiments (Figures S3–S5 and Table S1) revealed a high affinity of the TPE ligand for MGL, both the extracellular portion and CRD. 1S has a Kd of 67 and 250 nM for MGL CRD and full length, respectively. The fraction of active ligand (rA) was found to be <1 (Figure S6), suggesting that the formation of supramolecular clusters blocks access to a proportion of the ligand binding sites. Furthermore, the trivalent glycocluster 1S provides an unutilized phenol that can be harnessed to install a conjugation handle.

Synthesis of Trivalent MGL Ligand 1

The synthesis of 1 is summarized in Scheme 1.
The synthesis commenced from 2, which was prepared as described previously. (35) The direct tri-O-propargylation of 2 with propargyl bromide gave 3 in 34% yield. The propargylation gave also mono, di and tetra-O-propargylated products, however, this was still more efficient than a sequence involving the mono protection of an hydroxyl group of 2, followed by tripropargylation and then removal of the protecting group. (31) The phenolic compound 3 was then reacted with triethylene glycol (TEG) chloride, followed by treatment of the primary alcohol under Mitsunobu type reaction to obtain the azide 5, which was then reduced to the corresponding amine using tributylphosphine and was subsequently coupled with N-Fmoc-Asp-OtBu using EEDQ to give compound 7 in 85% yield. The S-GalNAc derivative I was synthesized as reported previously (30) and reacted with 7 via copper(I)-catalyzed 1,3-dipolar azide–alkyne cycloaddition. (31) The final acidic cleavage of the tert-butyl ester of 8 gave the desired compound TPEaa 1 for Fmoc-SPPS with an overall yield of 14%.

Synthesis of MUC1 Peptides, Conjugates with Trivalent S-GalNAc, and Vaccine Constructs to Target MGL

Since MUC1 is ubiquitously found on epithelial cell surfaces and tumor-associated glycoforms with an aberrant glycosylation pattern are overexpressed in many carcinomas, it is an attractive antigenic target for the development of cancer vaccines. Because the immune system has a natural tolerance for endogenous structures, MUC1-based vaccines employing the unmodified MUC1 TR sequence HGVT*S*APDT*RPAPGS*T*APPA (* represents possible glycosylation sites) as the antigen are only weakly immunogenic.
To enhance the immunogenicity, TACAs can be introduced into the MUC1 vaccine. A study on monoclonal antibodies suggested that the PDTR region is immunodominant in mice that were immunized with MUC1 vaccines. (36) Additionally, carbohydrate clusters, with the carbohydrate on adjacent Ser and Thr residues, were found to be the preferred epitopes of monoclonal antibodies. (37−40) With this in mind, we chose the MUC1 TR sequence glycosylated with the Tn-antigen in the PDT*R and the GS*T*A regions as antigen peptides. Although mucin-derived peptides can engage and crosslink B cell receptors, assigning them the unique or sole role as B cell epitopes is limiting as they may also contain T cell epitopes. For instance, the antigenic-dominant domain of MUC1 TR, which is included in the sequence SAPDT*RPAP, can complex with MHC class I (Kb) molecules and trigger the activation of cytotoxic T lymphocytes. (41) Furthermore, peptides can function as a shuttle to present carbohydrate antigens to T cells and generate carbohydrate-specific T cells that trigger a superior cellular immune response. (42) On the other hand, specific T follicular helper cells promote B cell class switching and memory formation. (43)
We planned the synthesis of two sets of peptides: the first set for in vitro and in cellulo testing and the second set designed for in vivo testing.
For the first set, a small library of control peptides and tumor-associated MUC1 TR glycopeptides carrying Tn-antigens were synthesized (Figure 1, peptides 9–14). One of the amino acids at the C-terminus was exchanged with a biotinylated lysine (KB) for peptide immobilization on streptavidin-coated ELISA plates and for fluorophore conjugation in flowcytometry and microscopy experiments. To test the influence of the TPEaa on MGL binding, ligand 1 was introduced to the N-terminus of peptides 12–14.

Figure 1

Figure 1. Summary of peptides 9–16 synthesized by SPPS.

High Resolution Image
Download MS PowerPoint Slide
In the second peptides set (Figure 1, peptides 15 and 16), we introduced the T helper epitope P30 (FNNFTVSFWLRVPKVSASHLE), derived from tetanus toxoid, to improve the presentation of exogenous glycopeptides on the MHC complex and activate T cells for in vivo testing. (44)
The tripartite vaccine 16 contains the Tn-modified MUC1 glycopeptide, the T helper epitope P30, and the high-affinity ligand 1 to enhance endocytosis by the C-type lectin MGL. Additionally, a corresponding control glycopeptide 15 lacking the MGL glycocluster ligand 1 was prepared.
Unglycosylated and Tn-modified MUC1 peptides were synthesized by automated SPPS according to a previously described protocol for glycopeptide synthesis. (45,46) Briefly, preloaded TentaGel R Trt Fmoc-Ala was used, and the standard Fmoc-protected amino acids as well as the biotinylated lysine were coupled using HBTU and HOBt. Glycosylated Ser and Thr amino acids (1.5 equiv) were pre-activated using the more reactive HATU and HOAt, manually added to the resin, and the coupling time was extended to 8 h. After complete assembly of peptides 14–16, a triethylene glycol spacer (Sp) was introduced to the N-terminus. The peptides were then further elongated with the P30 epitope by Fmoc-SPPS. We inserted TPEaa 1 by a final coupling step at the N-terminus and placed this furthest away from the resin, to maximize the coupling yield. To this end, TPEaa 1 (2 equiv) was manually pre-activated with HATU/HOAt and coupled to peptides 12–14 and 16 for 8 h. The obtained peptides were cleaved from the resin with simultaneous removal of acid-sensitive protecting groups on the amino acid side chains using TFA and TIPS. The O-acetyl protecting groups on the glycans were removed by transesterification in methanol using catalytic amounts of sodium methoxide, and compounds 9–16 were finally isolated after purification by C-18 preparative HPLC.

Recombinant MGL Receptor Binds to S-GalNAc MUC1 Glycopeptide Conjugates

To assess the ability of recombinant MGL to bind the synthesized peptides 9–14, we performed an ELISA using streptavidin-coated plates to which the biotinylated peptides 9–14 were immobilized. As reported in other affinity assessment assays, we varied the peptide concentration, while the lectin concentration was kept constant. (47,48)
To determine the optimal peptides and protein range to use in the assay, we performed several pilot titrations using a biotinylated polyacrylamide coupled-GalNAc polymer (PAA-GalNAC) (Figure S7).
The biotinylated peptides were coupled to a streptavidin-coated plate at twofold serial dilutions, which was then probed with recombinant MGL at a constant concentration tagged with a human IgG1Fc fragment and goat anti-human Fc-peroxidase was used for colorimetric detection.
The EC50 values derived from the ELISA binding studies with TA-MUC1 peptides are shown in Figure 2. Peptides 12–14 bearing the TPE α-S-GalNAc-glycocluster showed higher avidities than peptide 11 (Figure 2). MGL exhibited similar affinities for peptides 12–14, suggesting that MGL binds preferentially to the trivalent glycocluster over the Tn antigens on the MUC1 glycopeptide. A slight improvement in the EC50, from 3.90 nM for 13 to 2.65 nM for 14, was observed with the addition of a spacer separating the MUC1 peptide from the α-S-GalNAc-glycocluster. MGL binding to the unglycosylated MUC1 peptide 10 or to the tetanus toxoid P30 epitope 9 was not observed, confirming that glycosylation on the MUC1 backbone or the TPEaa ligand are crucial for MGL binding.

Figure 2

Figure 2. EC50 values for the Tn-MUC1 (glyco)peptides and their glycocluster-conjugates with MGL were obtained using ELISA assays of a dilution series of the immobilized peptides 9–14 and recombinant MGL: the biotinylated peptides 9–14 were coupled to a streptavidin-coated plate at twofold dilutions, which was then titrated with recombinant MGL-Fc. MGL binding was detected using peroxidase conjugated to a goat anti-human Fc Ab. Curve fitting is shown in the figure. For all the peptides, the R2 value of the curve fitting was above 0.98.

High Resolution Image
Download MS PowerPoint Slide

MGL Binds Vaccine Constructs and Mediates Antigen Uptake in Murine Bone Marrow-Derived Dendritic Cells

To assess the vaccine construct uptake by DCs via MGL, we isolated and cultured murine bone marrow-derived dendritic cells (BMDCs) and assessed the peptide uptake by flow cytometry. We allowed the peptide to bind to the cell surface at 4 °C and then labeled the constructs with streptavidin-PE. The cells were then incubated at 37 °C for 1 h to facilitate peptide internalization. Analysis of the mean fluorescence intensity (MFI) showed that the uptake of peptide 12 was fourfold larger compared to the uptake of peptides 10 and 11. The weak fluorescence increase of 12 in the 37 °C samples compared to the 4 °C samples suggests a receptor-mediated uptake mechanism (Figure 3A,B).

Figure 3

Figure 3. Flow cytometry analysis on BMDCs. (A) MFI of MHCII+CD11c+ cells relative to the peptides. BMDCs where incubated in the presence of the biotinylated peptides 10, 11, and 12 at 4 °C and then labeled with streptavidine-PE. (B) Peptides-associated fluorescence intensity histogram after incubation with BMDCs at 37 °C. (C) MFI relative to the MGL receptor on BMDC surface. BMDCs were treated with peptide 12 for 30 min at 37 °C. Error bars represent standard errors.

High Resolution Image
Download MS PowerPoint Slide
To verify whether the uptake was C-type lectin dependent, we sequestered calcium ions with EDTA. In the presence of the chelating agent, the binding was significantly diminished (Figure 3A,B). Furthermore, we used anti-MGL antibodies to evaluate the presence of MGL on the surface of the BMDCs post incubation with peptide 12. The amount of accessible MGL on the cell surface decreased by 48% in the samples treated with peptide 12 compared to the untreated cells (Figure 3C).
Peptide uptake was investigated by confocal microscopy. Peptide 10, serving as a negative control, and peptide 12 were incubated at 4 °C with BMDCs to allow binding to the MGL receptor. After a washing step, we labeled the bound peptides with streptavidin-Alexa 594. The cells incubated with the unglycosylated peptide 10 showed only unspecific binding (Figure 4A), while 12 showed high levels of cell surface binding (Figures 4B,C and S8). After incubation of the cells for 1 h at 37 °C, uptake of peptide 12 into the cells was detected. No uptake of 12 was observed at 4 °C, suggesting that the uptake is predominantly receptor mediated, thus excluding non-specific mechanisms of internalization. Again, no binding or uptake of peptide 10 was observed under any conditions, corroborating the observations and conclusions from flow cytometry experiments.

Figure 4

Figure 4. Comparison of fluorophore-labeled MUC1 peptides 10 and 12 by microscopy. BMDCs were treated with 5 μg/mL of the biotinylated peptides 10 and 12 at 4 °C for 1 h followed by washing and treatment with streptavidin-Alexa 594. (A,B) Localization of the peptides on the cells surface. (C) Localization of the peptides after incubation for 30 min and 1 h at 37 °C. Internalization was visualized by confocal microscopy.

High Resolution Image
Download MS PowerPoint Slide

S-GalNAc Glycocluster-MUC1 Conjugate Vaccine 16 Induces Higher IgG Antibody Titers

With the MUC1-p30 vaccine conjugates 15 and 16 in hand, we tested their ability to induce a strong and specific immune response in mice, in order to evaluate whether the trivalent MGL ligand S-GalNAc on the TPEaa glycocluster 16 would be beneficial compared to the non-functionalized derivative 15. Groups of four female C57BL/6J mice were immunized with conjugates 15 and 16 four times in 2-week intervals (days 0, 14, 28, and 42) with equal amounts of MUC1 peptides per injection. Serum samples were collected 5 days post-immunization, and antibody titers were determined by ELISA (Figure 5A). Glycopeptide 11 was conjugated to BSA, immobilized on ELISA plates, and probed with the antibody sera at twofold dilutions. The TPEaa vaccine 16 induced higher IgG antibody titers than the vaccine construct 15. At day 47, the IgG concentration in the sera of mice immunized with 16 is over sevenfold higher than that observed in mice immunized with 15 (Figure 5B). IgG subtyping analysis showed that vaccine 15 elicited IgG1, IgG2b, and IgG2c in a ratio 0.20:0.40:0.40, while vaccine 16 elicited IgG21, IgG2b, and IgG2c in a ratio 0.24:0.50:0.26 (Figure 5C). The antibody subtyping profile of 16, specifically the similar IgG1/IgG2c levels and higher IgG2b levels, suggest a mixed Th1/Th2 response. Previous work showed that the antibody subclasses profile of Qβ virus-like particles conjugated with 270 copies of STnMUC1(SAPDT*RPAP) (49,50) predominantly yielded a Th1-type profile. Construct 16 could be used in complementary settings to Qβ-(STnMUC1)270 when a mixed Th1/Th2 response is desired. (51) The produced IgG2b antibodies may activate antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity, and future studies will investigate further avenues and therapeutic potential in detail.

Figure 5

Figure 5. In vivo experiment immunization schedule and antibody analysis. (A) Experimental design: 2.5 nmol of construct per injection; day 0, immunization subcutaneous (s.c.) with complete Freund’s adjuvant (CFA); day s14, 28, and 42, immunization intraperitoneal (i.p.) with incomplete Freund’s Adjuvant (IFA); blood collection: 5 days post immunization. (B) IgG titers in the serum of mice immunized with constructs 15 and 16, respectively. The sera were collected at days 19, 33, and 47. (C) Total IgG and IgG subtypes from sera collected at day 47 from mice immunized with constructs 15 and 16. All ELISA measurements were performed on plates coated with BSA-(11)4-17 synthesized as reported in the Supporting Information. The p values are determined through two-way ANOVA with Šídák multiple comparison test using GraphPad Prism, ***p < 0.001; error bars indicate standard error.

High Resolution Image
Download MS PowerPoint Slide

Antibodies Induced by Glycocluster-MUC1 Conjugate Vaccine Bind to Native Breast Cancer Cells

Synthetic glycopeptide vaccines often employ glycans with lower degrees of complexity compared to native tumor antigens. These simpler constructs are not always able to induce specific antibodies that can recognize native tumors. (14) In order to determine the ability of the induced antibodies to recognize native cancer cells, sera collected at day 47 were incubated with T47D human breast cancer cells, which are known to overexpress aberrantly glycosylated MUC1. (52) Bound antibodies were labeled with a fluorescent anti-IgG secondary antibody for detection by flow cytometry. The MUC1-Tn antibodies induced after immunization with 15 and 16 recognized and bound to native breast cancer cells. As observed previously with the ELISA, vaccine construct 16 elicited a higher antibody titer compared to 15 (Figure 6).

Figure 6

Figure 6. Flow cytometry analysis of the specific recognition of breast tumor cells T47D by antibodies. (A) MFIs of binding of T47D-MUC1 cells by sera (1:100) of three mice (red, green, and blue curves) immunized with construct 16. The gray filled trace is from pre-immune serum. (B) Comparison of the MFIs of antibodies from sera of 15 and 16 (1:100). Error bars represent ±1 standard error from the mean of three biological replicates (n = 3). MUC1 expression on TD47 was confirmed by anti-MUC1 mAb HPMV (1:5 dilution) produced in-house. (53) The p-values were determined through a two-tailed t-test using GraphPad Prism, **p < 0.01; error bars indicate standard error.

High Resolution Image
Download MS PowerPoint Slide

Evaluation of Glycan Specificity of Antibodies Induced by 15 and 16

In order to evaluate the ability of vaccine conjugates 15 and 16 to induce strong and specific immune responses in vivo, the binding specificities of the polyclonal mouse sera obtained from immunization with vaccines 15 and 16 were determined using a synthetic MUC1 glycopeptide microarray library (Table S2). The microarrays were incubated with the obtained sera at a 1:1000 dilution. Bound antibodies were detected using a secondary Cy5-labeled goat anti-mouse IgG antibody. Microarray analysis showed similar binding patterns for antibodies elicited by vaccines 15 and 16, demonstrating that addition of the TPEaa does not affect antibody recognition (Figure 7). Additionally, higher antibody titers were again observed using construct 16. Furthermore, the antibodies exhibited preferences for glycosylation in the immune-dominant PDTR region over peptides glycosylated in the GVTS or GSTA regions.

Figure 7

Figure 7. Binding specificity of antibodies generated by vaccines 15 and 16 to mucin core Tn antigen, T antigen, and 1 and 3 glycopeptides. Binding of sera at 1:1000 dilution to an array of varied mucin peptide sequences, glycosylation sites, and glycan structures are shown. Mouse sera derived from 15 and 16 showed similar recognition patterns. However, higher binding to MUC1 glycopeptides was observed for mice immunized with 16. For complete figures, see S14 and S15. Bars represent standard error (n = 5).

High Resolution Image
Download MS PowerPoint Slide
Tn antigens and shorter linear mucin-type O-glycan core structures, typically found on cancer cells in patients with poor prognosis, (54,55) were better recognized by all sera compared to the peptides with extensive glycosylation with branched core 2 and core 4 structures, typically found on healthy cells. These latter diminished antibody recognitions of all antisera (Figure 8), suggesting that vaccine conjugates preferentially induced a tumor-specific humoral immune response.

Figure 8

Figure 8. Binding specificity of antibodies generated by vaccines 15 and 16 to mucin core 2 and 4 glycopeptides. Binding of sera at 1:1000 dilution to an array of varied mucin peptide sequences, glycosylation sites, and glycan structures are shown. Mouse sera derived from 15 and 16 showed preferential binding to tumor-associated mucin core. For complete figures, see S14 and S15. Bars represent standard error (n = 5).

High Resolution Image
Download MS PowerPoint Slide
Variations in selectivity among the sera from mice individuals for Tn antigens and other more extended glycan structures, as well as preferences for peptide epitopes glycosylated in the GSTA region, were observed. Importantly, these variations did not depend on the respective vaccine construct but could be attributed to the natural antibody production of the respective mouse.
No significant cross-reactivity with MUC4 and MUC5B glycopeptides carrying T- and Tn-antigens could be observed (Figures S16 and S17), indicating that the induced antibodies were specific for tumor-associated MUC1 glycopeptides rather than the glycan epitope only.

Conclusions

Click to copy section linkSection link copied!
We leveraged the increased affinity of TPE glycocluster constructs for MGL to develop a novel anti-cancer vaccine. We established a synthetic route to generate a synthetic tripartite vaccine containing a trivalent TPE glycocluster based on S-GalNAc to enhance MGL binding, a P30 T helper epitope and an antigenic Tn-modified MUC1 glycopeptide to induce tumor-specific antibodies in mice. ELISA experiments with recombinant MGL showed that MUC1-Tn glycopeptides displaying the trivalent TPE glycocluster were stronger binders compared to the corresponding MUC1-Tn glycopeptides without the trivalent S-GalNAc ligand. Additionally, flow cytometry analysis showed that the glycocluster facilitated increased internalization by DCs via the MGL receptor compared to the glycopeptide alone. The highest MGL binder was then introduced into the tripartite vaccine for immunization, leading to high titers of anti-Tn-MUC1 antibodies in vivo. The obtained antibodies were able to recognize MUC1-positive breast cancer cells. Microarray analysis showed that immunization with the vaccine construct bearing the multivalent S-GalNAc glycocluster led to increased antibody production without causing loss of selectivity for the target antigen. Furthermore, the antibodies exhibited preferences for short and linear mucin O-glycan core structures in the immune-dominant PDTR MUC1 region. Our results indicate that the vaccine construct 16 efficiently targets MGL, thus increasing antigen endocytosis by the MGL receptor and consequently eliciting a strong humoral response.
Thus, we created a structurally defined tripartite vaccine to enhance endocytosis and evaluated it in vitro and in vivo. The reported construct is based on a modular design and synthesis approach and offers the potential to transfer this modularity concept more widely to vaccine design. For example, different glycocluster ligands of high affinity for other DC lectins could be incorporated to investigate their properties, (56) or other glycopeptide constructs more efficiently targeted to DCs via MGL using the TPE ligand. Overall, our findings prove, for the first time, that the concept of targeting clinically relevant vaccine epitopes with a rationally designed MGL-ligand is applicable successfully in vivo, resulting in a boosted immune response.

Materials and Method

Click to copy section linkSection link copied!

Mouse Immunization

8 weeks old C57BL/6J female mice (Charles River) were housed and maintained in microisolator cages under specific pathogen-free conditions at the animal facility of Johannes Gutenberg-University according to institutionally approved protocols (permission was obtained from the Landesuntersuchungsamt Koblenz, 23 177-07/G 19-1-099). The mice received the first immunization s.c. with CFA on day 0 in a total volume of 50 μL into the right flank. The mice were immunized i.p. on days 14, 28, and 42 with IFA in a volume of 100 μL with a dose of 2.5 nmol per injection. Blood was collected from the tail vein five days post each vaccination, and serum was prepared by centrifugation for 15 min at 2500g.

Anti-Muc1 Antibody Quantification and Subtyping

ELISA was used for the analysis of induced anti-Muc1 antibodies in antiserum. 96-well MaxiSorp plates were coated by incubating overnight at 4 °C with 1 μg/mL glycopeptide 11 in BSA, washed three times with washing buffer (PBS containing 0.05% Tween-20), and blocked for 30 min at 37 °C with blocking buffer (washing buffer supplemented with 1% BSA). Dilutions of sera in blocking buffer were added and incubated for 2 h at 37 °C. The plates were washed three times with washing buffer and incubated with the different antibody subtypes IgG1, IgG2b (BD Pharmigen), IgG2a (PharMingen), IgG2c (Abcam), and IgG3 (BioLegend). After incubation and three washes, horseradish peroxidase-conjugated goat anti-mouse IgG (H + L) (Abcam) diluted 1:1000 in blocking buffer was added for 1 h at 37 °C. The plates were then washed three times and incubated for 10 min with 50 μL per well of 2,2′-azino-di-(3-ethylbenzothiazoline sulfonic acid substrate in citrate buffer (pH 4.4 and 1:4000) 30% H2O2. The reaction was quenched by adding 50 μL of 1 M aqueous sulfuric acid to each well. Plates were read at 410 nm using a Tecan Infinite F200 Pro microplate reader. For each IgG of the antiserum, optical densities were measured and the absorbance curves were fitted to obtain antibody titers. There were no IgGs detectable for the negative controls (untreated mice).

Anti-Muc1 Antibody Recognition of Breast Cancer Cells by Flow Cytometry

100.000 MUC1-positive T47D tumor cells (ATCC, Maryland USA) were incubated with 1 μL of the serum for 30 min at 4 °C in PBS. The cells were then washed two times with PBS and incubated with an Alexa Fluor 488 conjugated goat-anti-mouse-IgG secondary antibody (Invitrogen) and with fixable viability dye eFluor780 (Invitrogen) to exclude false-positive dead cells. The cells were washed two times and subsequently taken up and analyzed. Flow cytometry data were acquired on a Canto flow cytometer (BD Biosciences) and analyzed with FlowJo Software (FlowJoLLC).

Supporting Information

Click to copy section linkSection link copied!

The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/jacs.2c12843.

  • Experimental details and characterization of the synthetic antigens (PDF)

Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Author Information

Click to copy section linkSection link copied!
  • Corresponding Author
    • Adele Gabba - School of Biological and Chemical Sciences, University of Galway, University Rd., H91 TK33 Galway, Ireland;  Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany;  Orcidhttps://orcid.org/0000-0001-8240-6482;  Email: [email protected]
  • Authors
    • Riem Attariya - Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    • Sandra Behren - Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    • Christian Pett - Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    • Joost C. van der Horst - Department of Molecular Cell Biology and Immunology, Amsterdam UMC Location Vrije Universiteit Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands;  Amsterdam Institute for Infection and Immunity, Cancer Immunology1105 AZ Amsterdam, the Netherlands
    • Hajime Yurugi - Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    • Jin Yu - Glycosciences Laboratory, Imperial College London, W12 0NN London, U.K.
    • Moritz Urschbach - Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany
    • Juan Sabin - AFFINImeter Scientific & Development Team, Software 4 Science Developments, 15782 Santiago de Compostela, A Coruña, Spain;  Departamento de Física Aplicada, Facultad de Física, Universidad de Santiago de Compostela, 15705 Santiago de Compostela, Spain
    • Gabriel Birrane - Division of Experimental Medicine, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, United States;  Orcidhttps://orcid.org/0000-0002-1759-5499
    • Edgar Schmitt - Institute of Immunology, University Medical Center Mainz, Langenbeckstr.1, 55131 Mainz, Germany
    • Sandra J. van Vliet - Department of Molecular Cell Biology and Immunology, Amsterdam UMC Location Vrije Universiteit Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands;  Amsterdam Institute for Infection and Immunity, Cancer Immunology1105 AZ Amsterdam, the Netherlands
    • Pol Besenius - Department of Chemistry, Johannes Gutenberg University Mainz, Duesbergweg 10−14, 55128 Mainz, Germany;  Orcidhttps://orcid.org/0000-0001-7478-4459
    • Ulrika Westerlind - Department of Chemistry, Umeå University, KBC-Building, Linneaus väg 6, S-907 36 Umeå, Sweden
    • Paul V. Murphy - School of Biological and Chemical Sciences, University of Galway, University Rd., H91 TK33 Galway, Ireland;  SSPC, the Science Foundation Ireland Research Centre for Pharmaceuticals, University of Galway, University Rd., H91 TK33 Galway, Ireland;  Orcidhttps://orcid.org/0000-0002-1529-6540
  • Author Contributions

    S.J.v.V., P.B., U.W., and P.V.M. are joint senior authors.

  • Funding

    This work was supported by Science Foundation Ireland via grant number 16/IA/4419 to P.V.M., by the Irish Research Council via grant number GOIPG/2016/858 and EMBO short-term fellowship 7323 to A.G.. M.U., A.G., E.S., and P.B. acknowledge financial support from the Deutsche Forschungs-gemeinschaft (DFG) (CRC 1066) and the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (ERC CoG SUPRAVACC- 819856). U.W. acknowledges support from Kempe foundation (JCK-1819.1 and JCK-1819.2).

  • Notes
    The authors declare no competing financial interest.

Acknowledgments

Click to copy section linkSection link copied!

The authors acknowledge Prof. L.L. Kiessling and V. Lensch, MIT Chemistry, for the discussions and training on flow cytometry, Prof. Horst Kunz for the discussion, and Ozgur Cakici and Prof. Jerome Groopman, at the Beth Israel Deaconess Medical Center, for use of the facilities and ITC training.

References

Click to copy section linkSection link copied!

This article references 56 other publications.

  1. 1
    van Vliet, S. J.; Saeland, E.; van Kooyk, Y. Sweet Preferences of MGL: Carbohydrate Specificity and Function. Trends Immunol. 2008, 29, 83– 90,  DOI: 10.1016/j.it.2007.10.010
    Google Scholar
    1
    Sweet preferences of MGL: carbohydrate specificity and function
    van Vliet, Sandra J.; Saeland, Eirikur; van Kooyk, Yvette
    Trends in Immunology (2008), 29 (2), 83-90CODEN: TIRMAE; ISSN:1471-4906. (Elsevier B.V.)
    A review. C-type lectins play important roles in both innate and adaptive immune responses. In contrast to the mannose- or fucose-specific C-type lectins DC-SIGN and mannose receptor, the galactose-type lectins, of which only macrophage galactose-type lectin (MGL) is found within the immune system, are less well known. MGL is selectively expressed by immature dendritic cells and macrophages with elevated levels on tolerogenic or alternatively activated subsets. Human MGL has an exclusive specificity for rare terminal GalNAc structures, which are revealed on the tumor-assocd. mucin MUC1 and CD45 on effector T cells. These findings implicate MGL in the homeostatic control of adaptive immunity. The authors discuss here the functional similarities and differences between MGL orthologs and compare MGL to its closest homolog, the liver-specific asialoglycoprotein receptor (ASGP-R).
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXhslGitLc%253D&md5=a79c991afa6912a3da6a2f349682dcb6
  2. 2
    van Vliet, S. J.; van Liempt, E.; Saeland, E.; Aarnoudse, C. A.; Appelmelk, B.; Irimura, T.; Geijtenbeek, T. B. H.; Blixt, O.; Alvarez, R.; van Die, I.; van Kooyk, Y. Carbohydrate Profiling Reveals a Distinctive Role for the C-Type Lectin MGL in the Recognition of Helminth Parasites and Tumor Antigens by Dendritic Cells. Int. Immunol. 2005, 17, 661– 669,  DOI: 10.1093/intimm/dxh246
    Google Scholar
    2
    Carbohydrate profiling reveals a distinctive role for the C-type lectin MGL in the recognition of helminth parasites and tumor antigens by dendritic cells
    van Vliet, Sandra J.; van Liempt, Ellis; Saeland, Eirikur; Aarnoudse, Corlien A.; Appelmelk, Ben; Irimura, Tatsuro; Geijtenbeek, Teunis B. H.; Blixt, Ola; Alvarez, Richard; van Die, Irma; van Kooyk, Yvette
    International Immunology (2005), 17 (5), 661-669CODEN: INIMEN; ISSN:0953-8178. (Oxford University Press)
    Dendritic cells (DCs) are key to the maintenance of peripheral tolerance to self-antigens and the orchestration of an immune reaction to foreign antigens. C-type lectins, expressed by DCs, recognize carbohydrate moieties on antigens that can be internalized for processing and presentation. Little is known about the exact glycan structures on self-antigens and pathogens that are specifically recognized by the different C-type lectins and how this interaction influences DC function. The authors have analyzed the carbohydrate specificity of the human C-type lectin macrophage galactose-type lectin (MGL) using glycan microarray profiling and identified an exclusive specificity for terminal α- and β-linked GalNAc residues that naturally occur as parts of glycoproteins or glycosphingolipids. Specific glycan structures contg. terminal GalNAc moieties, expressed by the human helminth parasite Schistosoma mansoni as well as tumor antigens and a subset of gangliosides, were identified as ligands for MGL. The authors' results indicate an endogenous function for DC-expressed MGL in the clearance and tolerance to self-gangliosides, and in the pattern recognition of tumor antigens and foreign glycoproteins derived from helminth parasites.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXjslOqu70%253D&md5=8f8fc96e5d622e47338311d219077a3e
  3. 3
    Pirro, M.; Rombouts, Y.; Stella, A.; Neyrolles, O.; Burlet-Schiltz, O.; van Vliet, S. J.; de Ru, A. H.; Mohammed, Y.; Wuhrer, M.; van Veelen, P. A.; Hensbergen, P. J. Characterization of Macrophage Galactose-Type Lectin (MGL) Ligands in Colorectal Cancer Cell Lines. Biochim. Biophys. Acta, Gen. Subj. 2020, 1864, 129513,  DOI: 10.1016/j.bbagen.2020.129513
    Google Scholar
    3
    Characterization of Macrophage Galactose-type Lectin (MGL) ligands in colorectal cancer cell lines
    Pirro, Martina; Rombouts, Yoann; Stella, Alexandre; Neyrolles, Olivier; Burlet-Schiltz, Odile; van Vliet, Sandra J.; de Ru, Arnoud H.; Mohammed, Yassene; Wuhrer, Manfred; van Veelen, Peter A.; Hensbergen, Paul J.
    Biochimica et Biophysica Acta, General Subjects (2020), 1864 (4), 129513CODEN: BBGSB3; ISSN:0304-4165. (Elsevier B.V.)
    The Ca2+-dependent C-type lectin receptor Macrophage Galactose-type Lectin (MGL) is highly expressed by tolerogenic dendritic cells (DC) and macrophages. MGL exhibits a high binding specificity for terminal alpha- and beta-linked GalNAc residues found in Tn, sTn and LacdiNAc antigens. These glycan epitopes are often overexpressed in colorectal cancer (CRC), and, as such, MGL can be used to discriminate tumor from the corresponding healthy tissues. Moreover, the high expression of MGL ligands is assocd. with poor disease-free survival in stage III of CRC tumors. Nonetheless, the glycoproteins expressed by tumor cells that are recognized by MGL have hitherto remained elusive. Using a panel of three CRC cell lines (HCT116, HT29 and LS174T), recapitulating CRC diversity, we performed FACS staining and pull-down assays using a recombinant sol. form of MGL (and a mutant MGL as control) combined with mass spectrometry-based (glyco)proteomics. HCT116 and HT29, but not LS174T, are high MGL-binding CRC cell lines. On these cells, the major cell surface binding proteins are receptors (e.g. MET, PTK7, SORL1, PTPRF) and integrins (ITGB1, ITGA3). From these proteins, several N- and/or O-glycopeptides were identified, of which some carried either a LacdiNAc or Tn epitope. We have identified cell surface MGL-ligands on CRC cell lines. Advances in (glyco)proteomics have led to identification of candidate key mediators of immune-evasion and tumor growth in CRC.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhtFCktbw%253D&md5=d8254b636571303acda5e1c3491ea474
  4. 4
    Marcelo, F.; Supekar, N.; Corzana, F.; Van Der Horst, J. C.; Vuist, I. M.; Live, D.; Boons, G. J. P. H.; Smith, D. F.; Van Vliet, S. J. Identification of a Secondary Binding Site in Human Macrophage Galactose-Type Lectin by Microarray Studies: Implications for the Molecular Recognition of Its Ligands. J. Biol. Chem. 2019, 294, 1300– 1311,  DOI: 10.1074/jbc.ra118.004957
    Google Scholar
    4
    Identification of a secondary binding site in human macrophage galactose-type lectin by microarray studies: Implications for the molecular recognition of its ligands
    Marcelo, Filipa; Supekar, Nitin; Corzana, Francisco; van der Horst, Joost C.; Vuist, Ilona M.; Live, David; Boons, Geert-Jan P. H.; Smith, David F.; van Vliet, Sandra J.
    Journal of Biological Chemistry (2019), 294 (4), 1300-1311CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
    The human macrophage galactose-type lectin (MGL) is a C-type lectin characterized by a unique specificity for terminal GalNAc residues present in the tumor-assocd. Tn antigen (αGalNAc-Ser/Thr) and its sialylated form, the sialyl-Tn antigen. However, human MGL has multiple splice variants, and whether these variants have distinct ligand-binding properties is unknown. Here, using glycan microarrays, we compared the binding properties of the short MGL 6C (MGLshort) and the long MGL 6B (MGLlong) splice variants, as well as of a histidine-to-threonine mutant (MGLshort H259T). Although the MGLshort and MGLlong variants displayed similar binding properties on the glycan array, the MGLshort H259T mutant failed to interact with the sialyl-Tn epitope. As the MGLshort H259T variant could still bind a single GalNAc monosaccharide on this array, we next investigated its binding characteristics to Tn-contg. glycopeptides derived from the MGL ligands mucin 1 (MUC1), MUC2, and CD45. Strikingly, in the glycopeptide microarray, the MGLshort H259T variant lost high-affinity binding toward Tn-contg. glycopeptides, esp. at low probing concns. Moreover, MGLshort H259T was unable to recognize cancer-assocd. Tn epitopes on tumor cell lines. Mol. dynamics simulations indicated that in WT MGLshort, His259 mediates H bonds directly or engages the Tn-glycopeptide backbone through water mols. These bonds were lost in MGLshort H259T, thus explaining its lower binding affinity. Together, our results suggest that MGL not only connects to the Tn carbohydrate epitope, but also engages the underlying peptide via a secondary binding pocket within the MGL carbohydrate recognition domain contg. the His259 residue.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvVCjsL0%253D&md5=edab0eacc775c4e5b0eab684197e413a
  5. 5
    Zaal, A.; Li, R. J. E.; Lübbers, J.; Bruijns, S. C. M.; Kalay, H.; van Kooyk, Y.; van Vliet, S. J. Activation of the C-Type Lectin MGL by Terminal GalNAc Ligands Reduces the Glycolytic Activity of Human Dendritic Cells. Front. Immunol. 2020, 11, 305,  DOI: 10.3389/fimmu.2020.00305
    Google Scholar
    5
    Activation of the C-type lectin MGL by terminal GalNAc ligands reduces the glycolytic activity of human dendritic cells
    Zaal, Anouk; Eveline Li, R. J.; Luebbers, Joyce; Bruijns, Sven C. M.; Kalay, Hakan; van Kooyk, Yvette; van Vliet, Sandra J.
    Frontiers in Immunology (2020), 11 (), 305CODEN: FIRMCW; ISSN:1664-3224. (Frontiers Media S.A.)
    Many tumors display alterations in biosynthetic pathways of glycosylation, resulting in increased expression of specific tumor-assocd. glycan structures. Expression of these altered glycan structures is assocd. with metastasis and poor prognosis. Antigen presenting cells can recognize tumor-assocd. glycan structures, including the truncated O-glycan Tn antigen, via specific glycan receptors. Tn antigen-mediated activation of the C-type lectin MGL on dendritic cells induces regulatory T cells via the enhanced secretion of IL-10. Although these findings indicate that MGL engagement by glycan ligands can modulate immune responses, the impact of MGL ligation on dendritic cells is still not completely understood. Therefore, we employed RNA sequencing, GO term enrichment and pathway anal. on human monocyte-derived dendritic cells stimulated with two different MGL glycan ligands. Our analyses revealed a reduced expression of genes coding for key enzymes involved in the glycolysis pathway, TCA cycle, and oxidative phosphorylation. In concordance with this, extracellular flux anal. confirmed the decrease in glycolytic activity upon MGL triggering in human dendritic cells. To our knowledge, we are the first to report a diminished glycolytic activity of human dendritic cells upon C-type lectin stimulation. Overall, our findings highlight the impact of tumor-assocd. glycans on dendritic cell biol. and metab. and will increase our understanding on how glycans can shape immunity.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhvVarsr7E&md5=c34bc71b5c601783d553f49625e0451e
  6. 6
    Marcelo, F.; Garcia-Martin, F.; Matsushita, T.; Sardinha, J.; Coelho, H.; Oude-Vrielink, A.; Koller, C.; André, S.; Cabrita, E. J.; Gabius, H. J.; Nishimura, S. I.; Jiménez-Barbero, J.; Cañada, F. J. Delineating Binding Modes of Gal/GalNAc and Structural Elements of the Molecular Recognition of Tumor-Associated Mucin Glycopeptides by the Human Macrophage Galactose-Type Lectin. Chem.─Eur. J. 2014, 20, 16147– 16155,  DOI: 10.1002/chem.201404566
    Google Scholar
    6
    Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin
    Marcelo, Filipa; Garcia-Martin, Fayna; Matsushita, Takahiko; Sardinha, Joao; Coelho, Helena; Oude-Vrielink, Anneloes; Koller, Christiane; Andre, Sabine; Cabrita, Eurico J.; Gabius, Hans-Joachim; Nishimura, Shin-ichiro; Jimenez-Barbero, Jesus; Canada, F. Javier
    Chemistry - A European Journal (2014), 20 (49), 16147-16155CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
    The human macrophage galactose-type lectin (MGL) is a key physiol. receptor for the carcinoma-assocd. Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the mol. recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by satn. transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control expts. with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with mol. dynamics simulations and Corcema-ST to establish a 3D view on the mol. recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by addnl. hydrogen bonds and CH-π contacts involving exclusively the NHAc moiety.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXitVCiurfN&md5=c30d5d96f7c5d59672d097293659fd42
  7. 7
    Mortezai, N.; Behnken, H. N.; Kurze, A. K.; Ludewig, P.; Buck, F.; Meyer, B.; Wagener, C. Tumor-Associated Neu5Ac-Tn and Neu5Gc-Tn Antigens Bind to C-Type Lectin CLEC10A (CD301, MGL). Glycobiology 2013, 23, 844– 852,  DOI: 10.1093/glycob/cwt021
    Google Scholar
    7
    Tumor-associated Neu5Ac-Tn and Neu5Gc-Tn antigens bind to C-type lectin CLEC10A (CD301, MGL)
    Mortezai, Naghmeh; Behnken, Henning N.; Kurze, Anna-Katharina; Ludewig, Peter; Buck, Friedrich; Meyer, Bernd; Wagener, Christoph
    Glycobiology (2013), 23 (7), 844-852CODEN: GLYCE3; ISSN:0959-6658. (Oxford University Press)
    In human tumors, glycoproteins often exhibit abnormal glycosylation patterns, e.g. certain Lewis structures, TF antigen, Tn antigen and/or their sialylated forms, creating addnl. binding sites for glycoreceptors. In the present study, we have analyzed the carbohydrate specificity of the C-type lectin CLEC10A using glycan profiling by ELISA (ELISA). In addn. to the known ligands, we show binding to two tumor-assocd. antigens, namely Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn, with an affinity of CLEC10A in the micromolar range. Detailed analyses of the glycan-lectin interactions were carried out by surface plasmon resonance (SPR) and satn. transfer difference (STD) NMR. CLEC10A binds Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn with dissocn. consts. of 297 and 80 μM, resp., as detd. by SPR. Comparison of the STD NMR (NMR) binding epitopes of Tn and Neu5Acα2,6-Tn revealed a const. binding mode of the N-acetylgalactosamine moiety. This finding is in good agreement with binding studies of CLEC10A transfectomas, which show a well-defined interaction of transmembrane CLEC10A with 6-sialylated-Tn structures. Since both Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn together with the previously known Tn antigen are expressed in human tumors such as mammary carcinoma, the interaction with CLEC10A expressed by macrophages and dendritic cells could be of major functional significance in tumor progression.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXptFeruro%253D&md5=9c362e8087a962acf61ac6b5b8d959e2
  8. 8
    Bulteau, F.; Thépaut, M.; Henry, M.; Hurbin, A.; Vanwonterghem, L.; Vivès, C.; Le Roy, A.; Ebel, C.; Renaudet, O.; Fieschi, F.; Coll, J. L. Targeting Tn-Antigen-Positive Human Tumors with a Recombinant Human Macrophage Galactose C-Type Lectin. Mol. Pharm. 2022, 19, 235– 245,  DOI: 10.1021/acs.molpharmaceut.1c00744
    Google Scholar
    8
    Targeting Tn-Antigen-Positive Human Tumors with a Recombinant Human Macrophage Galactose C-Type Lectin
    Bulteau, Francois; Thepaut, Michel; Henry, Maxime; Hurbin, Amandine; Vanwonterghem, Laetitia; Vives, Corinne; Le Roy, Aline; Ebel, Christine; Renaudet, Olivier; Fieschi, Franck; Coll, Jean-Luc
    Molecular Pharmaceutics (2022), 19 (1), 235-245CODEN: MPOHBP; ISSN:1543-8384. (American Chemical Society)
    Alterations in glycosylation cause the emergence of tumor-assocd. carbohydrate antigens (TACAs) during tumorigenesis. Truncation of O-glycans reveals the Thomsen nouveau (Tn) antigen, an N-acetylgalactosamine (GalNAc) frequently attached to serine or threonine amino acids, that is accessible on the surface of cancer cells but not on healthy cells. Interestingly, GalNac can be recognized by macrophage galactose lectin (MGL), a type C lectin receptor expressed in immune cells. In this study, recombinant MGL fragments were tested in vitro for their cancer cell-targeting efficiency by flow cytometry and confocal microscopy and in vivo after administration of fluorescent MGL to tumor-bearing mice. Our results demonstrate the ability of MGL to target Tn-pos. human tumors without inducing toxicity. This outcome makes MGL, a fragment of a normal human protein, the first vector candidate for in vivo diagnosis and imaging of human tumors and, possibly, for therapeutic applications.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXislKks7rF&md5=53c31461c4343a34f3d81b59300b2609
  9. 9
    Cheever, M. A.; Allison, J. P.; Ferris, A. S.; Finn, O. J.; Hastings, B. M.; Hecht, T. T.; Mellman, I.; Prindiville, S. A.; Viner, J. L.; Weiner, L. M.; Matrisian, L. M. The Prioritization of Cancer Antigens: A National Cancer Institute Pilot Project for the Acceleration of Translational Research. Clin. Cancer Res. 2009, 15, 5323– 5337,  DOI: 10.1158/1078-0432.ccr-09-0737
    Google Scholar
    9
    The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research
    Cheever Martin A; Allison James P; Ferris Andrea S; Finn Olivera J; Hastings Benjamin M; Hecht Toby T; Mellman Ira; Prindiville Sheila A; Viner Jaye L; Weiner Louis M; Matrisian Lynn M
    Clinical cancer research : an official journal of the American Association for Cancer Research (2009), 15 (17), 5323-37 ISSN:.
    The purpose of the National Cancer Institute pilot project to prioritize cancer antigens was to develop a well-vetted, priority-ranked list of cancer vaccine target antigens based on predefined and preweighted objective criteria. An additional aim was for the National Cancer Institute to test a new approach for prioritizing translational research opportunities based on an analytic hierarchy process for dealing with complex decisions. Antigen prioritization involved developing a list of "ideal" cancer antigen criteria/characteristics, assigning relative weights to those criteria using pairwise comparisons, selecting 75 representative antigens for comparison and ranking, assembling information on the predefined criteria for the selected antigens, and ranking the antigens based on the predefined, preweighted criteria. Using the pairwise approach, the result of criteria weighting, in descending order, was as follows: (a) therapeutic function, (b) immunogenicity, (c) role of the antigen in oncogenicity, (d) specificity, (e) expression level and percent of antigen-positive cells, (f) stem cell expression, (g) number of patients with antigen-positive cancers, (h) number of antigenic epitopes, and (i) cellular location of antigen expression. None of the 75 antigens had all of the characteristics of the ideal cancer antigen. However, 46 were immunogenic in clinical trials and 20 of them had suggestive clinical efficacy in the "therapeutic function" category. These findings reflect the current status of the cancer vaccine field, highlight the possibility that additional organized efforts and funding would accelerate the development of therapeutically effective cancer vaccines, and accentuate the need for prioritization.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BD1Mrpslamsw%253D%253D&md5=bf8652477b44e40ca6da6b83f8ec471d
  10. 10
    Nath, S.; Mukherjee, P. MUC1: A Multifaceted Oncoprotein with a Key Role in Cancer Progression. Trends Mol. Med. 2014, 20, 332– 342,  DOI: 10.1016/j.molmed.2014.02.007
    Google Scholar
    10
    MUC1: a multifaceted oncoprotein with a key role in cancer progression
    Nath, Sritama; Mukherjee, Pinku
    Trends in Molecular Medicine (2014), 20 (6), 332-342CODEN: TMMRCY; ISSN:1471-4914. (Elsevier Ltd.)
    A review. The transmembrane glycoprotein Mucin 1 (MUC1) is aberrantly glycosylated and overexpressed in a variety of epithelial cancers, and plays a crucial role in progression of the disease. Tumor-assocd. MUC1 differs from the MUC1 expressed in normal cells with regard to its biochem. features, cellular distribution, and function. In cancer cells, MUC1 participates in intracellular signal transduction pathways and regulates the expression of its target genes at both the transcriptional and post-transcriptional levels. This review highlights the structural and functional differences that exist between normal and tumor-assocd. MUC1. We also discuss the recent advances made in the use of MUC1 as a biomarker and therapeutic target for cancer.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXksleqs7w%253D&md5=73e37d4a1e83b57e90078c2b4a4afbc0
  11. 11
    Springer, G. F. Immunoreactive T and Tn Epitopes in Cancer Diagnosis, Prognosis, and Immunotherapy. J. Mol. Med. 1997, 75, 594– 602,  DOI: 10.1007/s001090050144
    Google Scholar
    11
    Immunoreactive T and Tn epitopes in cancer diagnosis, prognosis, and immunotherapy
    Springer, Georg F.
    Journal of Molecular Medicine (Berlin) (1997), 75 (8), 594-602CODEN: JMLME8; ISSN:0946-2716. (Springer)
    A review with 74 refs. Aberrant glycosylation is a hallmark of cancer cells. The blood group precursors T (Thomsen-Friedenreich) and Tn epitopes are shielded in healthy and benign-diseased tissues but uncovered in approx. 90% of carcinomas. T and Tn glycoproteins are specific, autoimmunogenic pancarcinoma antigens. These antigens may also be found in neoplastic blood cells (and on LTV-II infected T lymphocytes). Fundamental chem. and phys. aspects of these glycoproteins of primary carcinomas are discussed first. Tn and T epitopes are cell and tissue adhesion mols., essential in invasion by and metastasis of carcinoma which includes adherent and proliferative phases. These properties are then delineated next, followed by consideration of pathophysiol. and clin. aspects of these antigens. Immunohistochem. studies of the extent of Tn antigen expression in primary breast carcinoma demonstrate its highly significant correlation with clinicopathol. tumor stages, and hence its value as a reliable prognosticator. On the other hand, there is no significant, prognostically useful assocn. between T antigen expression and clin. disease course. Everyone has "preexisting" anticarcinoma anti-Tn and anti-T antibodies, induced predominantly by the intestinal flora, while cellular immune responses to T and Tn epitopes are evoked only by carcinomas and some lymphomas. Carcinoma dedifferentiation leading to predominance of Tn over T epitopes is described, as is the role of Tn and T epitopes in very early, including preclin., carcinoma detection. The highest sensitivities in carcinoma detection are for preclin. and the earliest clin. stages. Obviously, preclin. carcinoma detection is of practical importance. T/anti-T tests detected preclin. carcinoma in 77% of 48 patients long (‾ 6 yr) before their biopsy/X-ray results became pos. There were no false predictions of carcinoma in 38 control persons with benign diseases (observation av. 4.8 yr). These findings open a novel window for both curative approaches and pathophysiol. studies. The autoimmunogenicity of carcinoma T/Tn antigen led us more than two decades ago to begin intradermal vaccination of patients with advanced breast carcinoma of stages IV-IIb, predominately after modified radical mastectomy and sometimes lumpectomy plus axillary dissection always followed by adjuvant radio/chemotherapy. The vaccine consists of human group O red blood cell membrane derived, HLA-free T/Tn antigen contg. as adjuvant Ca3(PO4)2 plus a trace of phosphoglycolipid A hyperantigen, i.e., S. typhi vaccine (USP), which itself has T and Tn specificities. Our efforts have now for up to 20 yr remained successful in a large majority of the 32 patients. All 32 patients survived at least 5 yr; 10-yr survival was statistically highly significantly improved (5-yr survival: P<1×10-7; 10-yr survival: P<1×10-5) compared to statistics of the United States National Cancer Institute. Because these vaccinations are successful, their extension to large populations with major types of carcinomas should be considered, and even immunol. carcinoma prevention may be contemplated.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXlvVKjtL8%253D&md5=0d81fbc8c7a7835e58872c3c707a2115
  12. 12
    Beckwith, D. M.; Cudic, M. Tumor-Associated O-Glycans of MUC1: Carriers of the Glyco-Code and Targets for Cancer Vaccine Design. Semin Immunol. 2020, 47, 101389,  DOI: 10.1016/j.smim.2020.101389
    Google Scholar
    12
    Tumor-associated O-glycans of MUC1: Carriers of the glyco-code and targets for cancer vaccine design
    Beckwith, Donella M.; Cudic, Mare
    Seminars in Immunology (2020), 47 (), 101389CODEN: SEIME2; ISSN:1044-5323. (Elsevier Ltd.)
    A review. The transformation from normal to malignant phenotype in human cancers is assocd. with aberrant cell-surface glycosylation. It has frequently been reported that MUC1, the heavily glycosylated cell-surface mucin, is altered in both, expression and glycosylation pattern, in human carcinomas of the epithelium. The presence of incomplete or truncated glycan structures, often capped by sialic acid, commonly known as tumor-assocd. carbohydrate antigens (TACAs), play a key role in tumor initiation, progression, and metastasis. Accumulating evidence suggests that expression of TACAs is assocd. with tumor escape from immune defenses. In this report, we will give an overview of the oncogenic functions of MUC1 that are exerted through TACA interactions with endogenous carbohydrate-binding proteins (lectins). These interactions often lead to creation of a pro-tumor microenvironment, favoring tumor progression and metastasis, and tumor evasion. In addn., we will describe current efforts in the design of cancer vaccines with special emphasis on synthetic MUC1 glycopeptide vaccines. Anal. of the key factors that govern structure-based design of immunogenic MUC1 glycopeptide epitopes are described. The role of TACA type, position, and d. on obsd. humoral and cellular immune responses is evaluated.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXlslemsQ%253D%253D&md5=42abcf8643445549c0fe38087b7b7640
  13. 13
    Gaidzik, N.; Westerlind, U.; Kunz, H. The Development of Synthetic Antitumour Vaccines from Mucin Glycopeptide Antigens. Chem. Soc. Rev. 2013, 42, 4421– 4442,  DOI: 10.1039/c3cs35470a
    Google Scholar
    13
    The development of synthetic antitumor vaccines from mucin glycopeptide antigens
    Gaidzik, Nikola; Westerlind, Ulrika; Kunz, Horst
    Chemical Society Reviews (2013), 42 (10), 4421-4442CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
    A review. Based on important cell-biol. and biochem. results concerning the structural difference between membrane glycoproteins of normal epithelial cells and epithelial tumor cells, tumor-assocd. glycopeptide antigens were chem. synthesized and structurally confirmed. Glycopeptide structures of the tandem repeat sequence of mucin MUC1 of epithelial tumor cells constitute the most promising tumor-assocd. antigens. In order to generate a sufficient immunogenicity of these endogenous structures, usually tolerated by the immune system, these synthetic glycopeptide antigens were conjugated to immune stimulating components: in fully synthetic 2-component vaccines either with T-cell peptide epitopes or with Toll-like receptor2 lipopeptide ligands or in 3-component vaccines with both these stimulants. Alternatively, the synthetic glycopeptide antigens were coupled to immune stimulating carrier proteins. In particular, MUC1 glycopeptide conjugates with Tetanus toxoid proved to be efficient vaccines inducing very strong immune responses in mice. The antibodies elicited with the fully synthetic vaccines showed selective recognition of the tumor-assocd. glycopeptides as was shown by neutralization and micro-array binding expts. After booster immunizations, most of the immune responses showed the installation of an immunol. memory. Immunization with fully synthetic 3-component vaccines induced immune reactions with therapeutic effects in terms of redn. of the tumor burden in mice or in killing of tumor cells in culture, while MUC1 glycopeptide-Tetanus toxoid vaccines elicited antibodies in mice which recognized tumor cells in human tumor tissues. The results achieved so far are considered to be promising for the development of an active immunization against tumors.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXms1Gitb0%253D&md5=038515ff3ac8ec3851429dbe8866a781
  14. 14
    Stergiou, N.; Urschbach, M.; Gabba, A.; Schmitt, E.; Kunz, H.; Besenius, P. The Development of Vaccines from Synthetic Tumor-Associated Mucin Glycopeptides and Their Glycosylation-Dependent Immune Response. Chem. Rec. 2021, 21, 3313– 3331,  DOI: 10.1002/tcr.202100182
    Google Scholar
    14
    The Development of Vaccines from Synthetic Tumor-Associated Mucin Glycopeptides and their Glycosylation-Dependent Immune Response
    Stergiou, Natascha; Urschbach, Moritz; Gabba, Adele; Schmitt, Edgar; Kunz, Horst; Besenius, Pol
    Chemical Record (2021), 21 (11), 3313-3331CODEN: CRHEAK; ISSN:1528-0691. (Wiley-VCH Verlag GmbH & Co. KGaA)
    A review. Tumor-assocd. carbohydrate antigens are overexpressed as altered-self in most common epithelial cancers. Their glycosylation patterns differ from those of healthy cells, functioning as an ID for cancer cells. Scientists have been developing anti-cancer vaccines based on mucin glycopeptides, yet the interplay of delivery system, adjuvant and tumor assocd. MUC epitopes in the induced immune response is not well understood. The current state of the art suggests that the identity, abundancy and location of the glycans on the MUC backbone are all key parameters in the cellular and humoral response. This review shares lessons learned by us in over two decades of research in glycopeptide vaccines. By bridging synthetic chem. and immunol., we discuss efforts in designing synthetic MUC1/4/16 vaccines and focus on the role of glycosylation patterns. We provide a brief introduction into the mechanisms of the immune system and aim to promote the development of cancer subunit vaccines.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXisFOiurzI&md5=83bfea0e1d565dc62594bc6566d06ae5
  15. 15
    Rowse, G. J.; Tempero, R. M.; VanLith, M. L.; Hollingsworth, M. A.; Gendler, S. J. Tolerance and Immunity to MUC1 in a Human MUC1 Transgenic Murine Model. Cancer Res. 1998, 58, 315– 321
    Google Scholar
    15
    Tolerance and immunity to MUC1 in a human MUC1 transgenic murine model
    Rowse, Gerald J.; Tempero, Richard M.; VanLith, Michelle L.; Hollingsworth, Michael A.; Gendler, Sandra J.
    Cancer Research (1998), 58 (2), 315-321CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
    The human epithelial mucin, MUC1, is a large transmembrane glycoprotein that is expressed on most simple epithelia. It is overexpressed and aberrantly glycosylated on many human epithelial tumors, including more than 90% of human breast cancers. MUC1 is of interest as an immunotherapy target because patients with breast, ovarian, and pancreatic cancers have T lymphocytes in their tumor-draining lymph nodes that can be induced to recognize and lyse MUC1-expressing tumor cells. We have produced a transgenic mouse model that expresses the human MUC1 mol. on an inbred C57Bl/6 background to investigate the effect of endogenous expression of MUC1 on the ability of mice to generate anti-tumor immunity to MUC1-expressing tumors. Transgenic mice expressed the human transgene in a pattern and level consistent with that obsd. in humans. Transgenic mice were tolerant to stimulation by MUC1 as evidenced by the ability of MUC1-expressing tumor cells to grow in these mice, whereas MUC1-expressing cells were eliminated from wild-type mice. Moreover, transgenic mice immunized with MUC1 peptides failed to exhibit Ig class switching to the IgG subtypes. These data suggest that endogenous expression of MUC1 protein by MUC1 transgenic mice induces T-cell tolerance to stimulation by MUC1. The transgenic mice will provide a useful model to investigate the mechanisms that regulate immunol. tolerance to tumor antigens and will facilitate the investigation of anti-MUC1 immunotherapy formulations.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1cXntF2gtQ%253D%253D&md5=4dd48bf8e125ca46d65dca946d6b3171
  16. 16
    Brown, G. D.; Willment, J. A.; Whitehead, L. C-type lectins in immunity and homeostasis. Nat. Rev. Immunol. 2018, 18, 374– 389,  DOI: 10.1038/s41577-018-0004-8
    Google Scholar
    16
    C-type lectins in immunity and homeostasis
    Brown, Gordon D.; Willment, Janet A.; Whitehead, Lauren
    Nature Reviews Immunology (2018), 18 (6), 374-389CODEN: NRIABX; ISSN:1474-1733. (Nature Research)
    A review. The C-type lectins are a superfamily of proteins that recognize a broad repertoire of ligands and that regulate a diverse range of physiol. functions. Most research attention has focused on the ability of C-type lectins to function in innate and adaptive antimicrobial immune responses, but these proteins are increasingly being recognized to have a major role in autoimmune diseases and to contribute to many other aspects of multicellular existence. Defects in these mols. lead to developmental and physiol. abnormalities, as well as altered susceptibility to infectious and non-infectious diseases. In this Review, we present an overview of the roles of C-type lectins in immunity and homeostasis, with an emphasis on the most exciting recent discoveries.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhtFWqs7jI&md5=9da9ec71ab6fb3262f297cff160aa72d
  17. 17
    Ilarregui, J. M.; Kooij, G.; Rodríguez, E.; Van Der Pol, S. M. A.; Koning, N.; Kalay, H.; Van Der Horst, J. C.; Van Vliet, S. J.; García-Vallejo, J. J.; De Vries, H. E.; Van Kooyk, Y. Macrophage Galactose-Type Lectin (MGL) Is Induced on M2 Microglia and Participates in the Resolution Phase of Autoimmune Neuroinflammation. J. Neuroinflammation 2019, 16, 130– 214,  DOI: 10.1186/s12974-019-1522-4
    Google Scholar
    17
    Macrophage galactose-type lectin (MGL) is induced on M2 microglia and participates in the resolution phase of autoimmune neuroinflammation
    Ilarregui Juan M; Rodriguez Ernesto; Koning Nathalie; Kalay Hakan; van der Horst Joost C; van Vliet Sandra J; Garcia-Vallejo Juan J; van Kooyk Yvette; Kooij Gijs; van der Pol Susanne M A; de Vries Helga E
    Journal of neuroinflammation (2019), 16 (1), 130 ISSN:.
    BACKGROUND: Multiple sclerosis (MS) involves a misdirected immune attack against myelin in the brain and spinal cord, leading to profound neuroinflammation and neurodegeneration. While the mechanisms of disease pathogenesis have been widely studied, the suppression mechanisms that lead to the resolution of the autoimmune response are still poorly understood. Here, we investigated the role of the C-type lectin receptor macrophage galactose-type lectin (MGL), usually expressed on tolerogenic antigen-presenting cells (APCs), as a negative regulator of autoimmune-driven neuroinflammation. METHODS: We used in silico, immunohistochemical, immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR) and flow cytometry analysis to explore the expression and functionality of MGL in human macrophages and microglia, as well as in MS post-mortem tissue. In vitro, we studied the capacity of MGL to mediate apoptosis of experimental autoimmune encephalomyelitis (EAE)-derived T cells and mouse CD4(+) T cells. Finally, we evaluated in vivo and ex vivo the immunomodulatory potential of MGL in EAE. RESULTS: MGL plays a critical role in the resolution phase of EAE as MGL1-deficient (Clec10a(-/-)) mice showed a similar day of onset but experienced a higher clinical score to that of WT littermates. We demonstrate that the mouse ortholog MGL1 induces apoptosis of autoreactive T cells and diminishes the expression of pro-inflammatory cytokines and inflammatory autoantibodies. Moreover, we show that MGL1 but not MGL2 induces apoptosis of activated mouse CD4(+) T cells in vitro. In human settings, we show that MGL expression is increased in active MS lesions and on alternatively activated microglia and macrophages which, in turn, induces the secretion of the immunoregulatory cytokine IL-10, underscoring the clinical relevance of this lectin. CONCLUSIONS: Our results show a new role of MGL-expressing APCs as an anti-inflammatory mechanism in autoimmune neuroinflammation by dampening pathogenic T and B cell responses, uncovering a novel clue for neuroprotective therapeutic strategies with relevance for in MS clinical applications.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3Mzhs1Gqtg%253D%253D&md5=431c7d27f54f82c5dff177e464034b71
  18. 18
    Costa, M.; da Costa, V.; Lores, P.; Landeira, M.; Rodríguez-Zraquia, S. A.; Festari, M. F.; Freire, T. Macrophage Gal/GalNAc Lectin 2 (MGL2)+ Peritoneal Antigen Presenting Cells during Fasciola Hepatica Infection Are Essential for Regulatory T Cell Induction. Sci. Rep. 2022, 12, 17661– 17712,  DOI: 10.1038/s41598-022-21520-w
    Google Scholar
    18
    Macrophage Gal/GalNAc lectin 2 (MGL2)+ peritoneal antigen presenting cells during Fasciola hepatica infection are essential for regulatory T cell induction
    Costa, Monique; da Costa, Valeria; Lores, Pablo; Landeira, Mercedes; Rodriguez-Zraquia, Santiago A.; Festari, Maria Florencia; Freire, Teresa
    Scientific Reports (2022), 12 (1), 17661CODEN: SRCEC3; ISSN:2045-2322. (Nature Portfolio)
    Fasciola hepatica, one of the agents that causes fasciolosis, modulates the host immune system to allow parasite survival in the host. F. hepatica expresses carbohydrate-contg. glycoconjugates that are decoded by C-type lectin receptors, such as Dectin-1, mannose receptor, DC-SIGN and MGL, that are mainly present on myeloid antigen presenting cells (APCs) and can mediate immunoregulatory properties on T cells. In particular, Macrophage Gal/GalNAc lectin 2 (MGL2) expands modified Th2 immune responses, while suppressing Th1 polarization, upon recognition of GalNAc-glycosylated parasite components. In this study, by using MGL2-DTR transgenic mice that encode human diphtheria toxin receptor in MGL2+ cells, we demonstrate the role of peritoneal APCs during F. hepatica infection in favoring parasite survival. This process might be mediated by the induction of splenic Tregs in vivo, since the depletion of MGL2+ cells conferred mice with partial resistance to the infection and abrogated the increase of CD4+/CD25+ FoxP3+ Tregs induced by the parasite. Therefore, MGL2+ cells are crit. determinants of F. hepatica infection and could constitute immune checkpoints to control parasite infection.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XislaksLzP&md5=8bf1f3ef110b3c9652c20404bec5ca1d
  19. 19
    Pace, A.; Scirocchi, F.; Napoletano, C.; Zizzari, I. G.; D’angelo, L.; Santoro, A.; Nuti, M.; Rahimi, H.; Rughetti, A. Glycan-Lectin Interactions as Novel Immunosuppression Drivers in Glioblastoma. Int. J. Mol. Sci. 2022, 23, 6312,  DOI: 10.3390/ijms23116312
    Google Scholar
    19
    Glycan-Lectin Interactions as Novel Immunosuppression Drivers in Glioblastoma
    Pace, Angelica; Scirocchi, Fabio; Napoletano, Chiara; Zizzari, Ilaria Grazia; D'Angelo, Luca; Santoro, Antonio; Nuti, Marianna; Rahimi, Hassan; Rughetti, Aurelia
    International Journal of Molecular Sciences (2022), 23 (11), 6312CODEN: IJMCFK; ISSN:1422-0067. (MDPI AG)
    A review. Despite diagnostic and therapeutic improvements, glioblastoma (GB) remains one of the most threatening brain tumor in adults, underlining the urgent need of new therapeutic targets. Lectins are glycan-binding proteins that regulate several biol. processes through the recognition of specific sugar motifs. Lectins and their ligands are found on immune cells, endothelial cells and, also, tumor cells, pointing out a strong correlation among immunity, tumor microenvironment and vascularization. In GB, altered glycans and lectins contribute to tumor progression and immune evasion, shaping the tumor-immune landscape promoting immunosuppressive cell subsets, such as myeloid-derived suppressor cells (MDSCs) and M2-macrophages, and affecting immunoeffector populations, such as CD8+ T cells and dendritic cells (DCs). Here, we discuss the latest knowledge on the immune cells, immune related lectin receptors (C-type lectins, Siglecs, galectins) and changes in glycosylation that are involved in immunosuppressive mechanisms in GB, highlighting their interest as possible novel therapeutical targets.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XhsFWru73N&md5=4b611e66666a11371dab01a910cc502d
  20. 20
    da Costa, V.; van Vliet, S. J.; Carasi, P.; Frigerio, S.; García, P. A.; Croci, D. O.; Festari, M. F.; Costa, M.; Landeira, M.; Rodríguez-Zraquia, S. A.; Cagnoni, A. J.; Cutine, A. M.; Rabinovich, G. A.; Osinaga, E.; Mariño, K. V.; Freire, T. The Tn Antigen Promotes Lung Tumor Growth by Fostering Immunosuppression and Angiogenesis via Interaction with Macrophage Galactose-Type Lectin 2 (MGL2). Cancer Lett. 2021, 518, 72– 81,  DOI: 10.1016/j.canlet.2021.06.012
    Google Scholar
    20
    The Tn antigen promotes lung tumor growth by fostering immunosuppression and angiogenesis via interaction with Macrophage Galactose-type lectin 2 (MGL2)
    da Costa, Valeria; van Vliet, Sandra J.; Carasi, Paula; Frigerio, Sofia; Garcia, Pablo A.; Croci, Diego O.; Festari, Maria Florencia; Costa, Monique; Landeira, Mercedes; Rodriguez-Zraquia, Santiago A.; Cagnoni, Alejandro J.; Cutine, Anabela M.; Rabinovich, Gabriel A.; Osinaga, Eduardo; Marino, Karina V.; Freire, Teresa
    Cancer Letters (New York, NY, United States) (2021), 518 (), 72-81CODEN: CALEDQ; ISSN:0304-3835. (Elsevier Inc.)
    Tn is a tumor-assocd. carbohydrate antigen that constitutes both a diagnostic tool and an immunotherapeutic target. It originates from interruption of the mucin O-glycosylation pathway through defects involving, at least in part, alterations in core-1 synthase activity, which is highly dependent on Cosmc, a folding chaperone. Tn antigen is recognized by the Macrophage Galactose-type Lectin (MGL), a C-type lectin receptor present on dendritic cells and macrophages. Specific interactions between Tn and MGL shape anti-tumoral immune responses by regulating several innate and adaptive immune cell programs. In this work, we generated and characterized a variant of the lung cancer murine cell line LL/2 that expresses Tn by mutation of the Cosmc chaperone gene (Tn+ LL/2). We confirmed Tn expression by lectin glycophenotyping and specific anti-Tn antibodies, verified abrogation of T-synthase activity in these cells, and confirmed its recognition by the murine MGL2 receptor. Interestingly, Tn+ LL/2 cells were more aggressive in vivo, resulting in larger and highly vascularized tumors than those generated from wild type Tn- LL/2 cells. In addn., Tn+ tumors exhibited an increase in CD11c+ F4/80+ cells with high expression of MGL2, together with an augmented expression of IL-10 in infiltrating CD4+ and CD8+ T cells. Importantly, this immunosuppressive microenvironment was dependent on the presence of MGL2+ cells, since depletion of these cells abrogated tumor growth, vascularization and recruitment of IL-10+ T cells. Altogether, our results suggest that expression of Tn in tumor cells and its interaction with MGL2-expressing CD11c+F4/80+ cells promote immunosuppression and angiogenesis, thus favoring tumor progression.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhtlKrtbfP&md5=3dcc08a15f2b731d5a4c66d19ae857a1
  21. 21
    Freire, T.; Lo-Man, R.; Bay, S.; Leclerc, C. Tn Glycosylation of the MUC6 Protein Modulates Its Immunogenicity and Promotes the Induction of Th17-Biased T Cell Responses. J. Biol. Chem. 2011, 286, 7797– 7811,  DOI: 10.1074/jbc.m110.209742
    Google Scholar
    21
    Tn Glycosylation of the MUC6 Protein Modulates its Immunogenicity and Promotes the Induction of Th17-biased T Cell Responses
    Freire, Teresa; Lo-Man, Richard; Bay, Sylvie; Leclerc, Claude
    Journal of Biological Chemistry (2011), 286 (10), 7797-7811CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
    The Tn antigen (α-GalNAc-O-Ser/Thr) is one of the most specific human cancer-assocd. structures. This antigen, together with mucins, the major carriers of O-glycosylated tumor antigens in adenocarcinomas, are being evaluated as anti-cancer immunotherapeutic targets. In particular, the MUC6 protein, which is normally expressed only in gastric tissues, has been detected in intestinal, pulmonary, colorectal, and breast carcinomas. To develop anti-cancer vaccines based on the Tn antigen, the authors produced MUC6 proteins with different Tn d. by using mixts. of recombinant ppGalNAc-T1, -T2, and -T7. The obtained glycoproteins were characterized and analyzed for their immunol. properties, as compared with the non-glycosylated MUC6. The authors show that these various MUC6:Tn glycoproteins were well recognized by both MUC6 and Tn-specific antibodies. However, Tn glycosylation of the MUC6 protein strongly affected their immunogenicity by partially abrogating Th1 cell responses, and promoting IL-17 responses. Moreover, the non-glycosylated MUC6 was more efficiently presented than MUC6:Tn glycoproteins to specific T CD4+ hybridomas, suggesting that Tn glycosylation may affect MUC6 processing or MHC binding of the processed peptides. In conclusion, the results indicate that Tn glycosylation of the MUC6 protein strongly affects its B and T cell immunogenicity, and might favor immune escape of tumor cells.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXislylsL8%253D&md5=fd44b4f44acf8d5e5ac975a79b421a65
  22. 22
    Freire, T.; Zhang, X.; Dériaud, E.; Ganneau, C.; Vichier-Guerre, S.; Azria, E.; Launay, O.; Lo-Man, R.; Bay, S.; Leclerc, C. Glycosidic Tn-Based Vaccines Targeting Dermal Dendritic Cells Favor Germinal Center B-Cell Development and Potent Antibody Response in the Absence of Adjuvant. Blood 2010, 116, 3526– 3536,  DOI: 10.1182/blood-2010-04-279133
    Google Scholar
    22
    Glycosidic Tn-based vaccines targeting dermal dendritic cells favor germinal center B-cell development and potent antibody response in the absence of adjuvant
    Freire, Teresa; Zhang, Xiaoming; Deriaud, Edith; Ganneau, Christelle; Vichier-Guerre, Sophie; Azria, Elie; Launay, Odile; Lo-Man, Richard; Bay, Sylvie; Leclerc, Claude
    Blood (2010), 116 (18), 3526-3536CODEN: BLOOAW; ISSN:0006-4971. (American Society of Hematology)
    In vivo targeting of C-type lectin receptors is an effective strategy for increasing antigen uptake and presentation by dendritic cells (DCs). To induce efficient immune response, glycosylated tumor-assocd. Tn antigens were used to target DCs through binding to macrophage galactose-type lectin (MGL). The capacity of Tn-glycosylated antigens-and the multiple antigenic glycopeptide Tn3 therapeutic candidate vaccine-to target mouse and human MGL+ DCs are demonstrated, esp. regarding dermal DCs. In mice, MGL+ CD103- dermal DCs efficiently captured and processed glycosylated Tn antigen in vivo, inducing a potent major histocompatibility complex (MHC) class II-restricted T-cell response. Intradermal immunization with Tn-glycopeptides induced high levels of Th2 cytokines-even in the presence of unmethylated cytosine-phosphate-guanosine-and was assocd. with increased expansion of the germinal center B-cell population. Therefore, MGL acts as an efficient endocytic antigen receptor on dermal DCs in vivo, able to prime Tn-specific T- and B-cell responses. Moreover, even in the absence of adjuvant, immunization with this glycosidic Tn-based vaccine induced high levels of anti-Tn antibody responses, recognizing human tumor cells. In vivo DC-targeting strategies, based on Tn-MGL interactions, constitute a promising strategy for enhancing antigen presentation and inducing potent antibody response.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhsVKksb7J&md5=0da063b69b6371bdb6d5f4687b13ce49
  23. 23
    Rosenbaum, P.; Artaud, C.; Bay, S.; Ganneau, C.; Campone, M.; Delaloge, S.; Gourmelon, C.; Loirat, D.; Medioni, J.; Pein, F.; Sablin, M. P.; Tredan, O.; Varga, A.; Leclerc, C. The Fully Synthetic Glycopeptide MAG-Tn3 Therapeutic Vaccine Induces Tumor-Specific Cytotoxic Antibodies in Breast Cancer Patients. Cancer Immunol. Immunother. 2020, 69, 703– 716,  DOI: 10.1007/s00262-020-02503-0
    Google Scholar
    23
    The fully synthetic glycopeptide MAG-Tn3 therapeutic vaccine induces tumor-specific cytotoxic antibodies in breast cancer patients
    Rosenbaum, Pierre; Artaud, Cecile; Bay, Sylvie; Ganneau, Christelle; Campone, Mario; Delaloge, Suzette; Gourmelon, Carole; Loirat, Delphine; Medioni, Jacques; Pein, Francois; Sablin, Marie-Paule; Tredan, Olivier; Varga, Andrea; Leclerc, Claude
    Cancer Immunology Immunotherapy (2020), 69 (5), 703-716CODEN: CIIMDN; ISSN:0340-7004. (Springer)
    Cancer is one of the main causes of mortality worldwide and a major public health concern. Among various strategies, therapeutic vaccines have been developed to stimulate anti-tumoral immune responses. However, in spite of extensive studies, this approach suffers from a lack of efficacy. We designed the MAG-Tn3 vaccine, aiming to induce antibody responses against Tn, a tumor-assocd. carbohydrate antigen. The fully synthetic glycopeptide vaccine MAG-Tn3 is composed of four arms built on three adjacent Tn moieties assocd. with the tetanus toxin-derived peptide TT830-844 CD4+ T-cell epitope. This promiscuous CD4+ T-cell epitope can bind to a wide range of HLA-DRB mols. and is thus expected to activate CD4+ T-cell responses in a large part of the human population. The MAG-Tn3 vaccine was formulated with the GSK-proprietary immunostimulant AS15, composed of CpG7909, MPL, and QS21, which has been shown to stimulate both innate and humoral responses, in addn. to being well tolerated. The first results of phase I clin. trial demonstrated that all vaccinated patients developed high levels of Tn-specific antibodies. Moreover, these antibodies specifically recognized Tn-expressing human tumor cells and killed them through a complement-dependent cytotoxicity mechanism. Overall, this study establishes, for the first time, the capacity of a fully synthetic glycopeptide cancer vaccine to induce specific immune responses in humans.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjtlajur0%253D&md5=446ca7adcafd30823c3a3158da8c9b2e
  24. 24
    Laubreton, D.; Bay, S.; Sedlik, C.; Artaud, C.; Ganneau, C.; Dériaud, E.; Viel, S.; Puaux, A. L.; Amigorena, S.; Gérard, C.; Lo-Man, R.; Leclerc, C. The Fully Synthetic MAG-Tn3 Therapeutic Vaccine Containing the Tetanus Toxoid-Derived TT830-844 Universal Epitope Provides Anti-Tumor Immunity. Cancer Immunol. Immunother. 2016, 65, 315– 325,  DOI: 10.1007/s00262-016-1802-0
    Google Scholar
    24
    The fully synthetic MAG-Tn3 therapeutic vaccine containing the tetanus toxoid-derived TT830-844 universal epitope provides anti-tumor immunity
    Laubreton, Daphne; Bay, Sylvie; Sedlik, Christine; Artaud, Cecile; Ganneau, Christelle; Deriaud, Edith; Viel, Sophie; Puaux, Anne-Laure; Amigorena, Sebastian; Gerard, Catherine; Lo-Man, Richard; Leclerc, Claude
    Cancer Immunology Immunotherapy (2016), 65 (3), 315-325CODEN: CIIMDN; ISSN:0340-7004. (Springer)
    Malignant transformations are often assocd. with aberrant glycosylation processes that lead to the expression of new carbohydrate antigens at the surface of tumor cells. Of these carbohydrate antigens, the Tn antigen is particularly highly expressed in many carcinomas, esp. in breast carcinoma. We designed MAG-Tn3, a fully synthetic vaccine based on three consecutive Tn moieties that are O-linked to a CD4+ T cell epitope, to induce anti-Tn antibody responses that could be helpful for therapeutic vaccination against cancer. To ensure broad coverage within the human population, the tetanus toxoid-derived peptide TT830-844 was selected as a T-helper epitope because it can bind to various HLA-DRB mols. We showed that the MAG-Tn3 vaccine, which was formulated with the GSK proprietary immunostimulant AS15 and designed for human cancer therapy, is able to induce an anti-Tn antibody response in mice of various H-2 haplotypes, and this response correlates with the ability to induce a specific T cell response against the TT830-844 peptide. The universality of the TT830-844 peptide was extended to new H-2 and HLA-DRB mols. that were capable of binding this T cell epitope. Finally, the MAG-Tn3 vaccine was able to induce anti-Tn antibody responses in cynomolgus monkeys, which targeted Tn-expressing tumor cells and mediated tumor cell death both in vitro and in vivo. Thus, MAG-Tn3 is a highly promising anticancer vaccine that is currently under evaluation in a phase I clin. trial.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XisFSmsr4%253D&md5=913211d77c205902ac2a681ce07698a5
  25. 25
    Eggink, L. L.; Roby, K. F.; Cote, R.; Kenneth Hoober, J. An Innovative Immunotherapeutic Strategy for Ovarian Cancer: CLEC10A and Glycomimetic Peptides. J. Immunother. Cancer 2018, 6, 28,  DOI: 10.1186/s40425-018-0339-5
    Google Scholar
    25
    An innovative immunotherapeutic strategy for ovarian cancer: CLEC10A and glycomimetic peptides
    Eggink Laura L; Cote Robert; Kenneth Hoober J; Roby Katherine F
    Journal for immunotherapy of cancer (2018), 6 (1), 28 ISSN:.
    BACKGROUND: Receptors specific for the sugar N-acetylgalactosamine (GalNAc) include the human type II, C-type lectin receptor macrophage galactose-type lectin/C-type lectin receptor family member 10A (MGL/CLEC10A/CD301) that is expressed prominently by human peripheral immature dendritic cells, dendritic cells in the skin, alternatively-activated (M2a) macrophages, and to lesser extents by several other types of tissues. CLEC10A is an endocytic receptor on antigen-presenting cells and has been proposed to play an important role in maturation of dendritic cells and initiation of an immune response. In this study, we asked whether a peptide that binds in the GalNAc-binding site of CLEC10A would serve as an effective tool to activate an immune response against ovarian cancer. METHODS: A 12-mer sequence emerged from a screen of a phage display library with a GalNAc-specific lectin. The peptide, designated svL4, and a shorter peptide consisting of the C-terminal 6 amino acids, designated sv6D, were synthesized as tetravalent structures based on a tri-lysine core. In silico and in vitro binding assays were developed to evaluate binding of the peptides to GalNAc-specific receptors. Endotoxin-negative peptide solutions were administered by subcutaneous injection and biological activity of the peptides was determined by secretion of cytokines and the response of peritoneal immune cells in mice. Anti-cancer activity was studied in a murine model of ovarian cancer. RESULTS: The peptides bound to recombinant human CLEC10A with high avidity, with half-maximal binding in the low nanomolar range. Binding to the receptor was Ca(2+)-dependent. Subcutaneous injection of low doses of peptides into mice on alternate days resulted in several-fold expansion of populations of mature immune cells within the peritoneal cavity. Peptide sv6D effectively suppressed development of ascites in a murine ovarian cancer model as a monotherapy and in combination with the chemotherapeutic drug paclitaxel or the immunotherapeutic antibody against the receptor PD-1. Toxicity, including antigenicity and release of cytotoxic levels of cytokines, was not observed. CONCLUSION: sv6D is a functional ligand for CLEC10A and induces maturation of immune cells in the peritoneal cavity. The peptide caused a highly significant extension of survival of mice with implanted ovarian cancer cells with a favorable toxicity and non-antigenic profile.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1MjjslCrtA%253D%253D&md5=cd7dab036f346f98d7bccb199c31e183
  26. 26
    Heger, L.; Balk, S.; Lühr, J. J.; Heidkamp, G. F.; Lehmann, C. H. K.; Hatscher, L.; Purbojo, A.; Hartmann, A.; Garcia-Martin, F.; Nishimura, S. I.; Cesnjevar, R.; Nimmerjahn, F.; Dudziak, D. CLEC10A Is a Specific Marker for Human CD1c+dendritic Cells and Enhances Their Toll-like Receptor 7/8-Induced Cytokine Secretion. Front. Immunol. 2018, 9, 744,  DOI: 10.3389/fimmu.2018.00744
    Google Scholar
    26
    CLEC10A is a specific marker for human CD1c+ dendritic cells and enhances their toll-like receptor 7/8-induced cytokine secretion
    Heger, Lukas; Balk, Silke; Luehr, Jennifer J.; Heidkamp, Gordon F.; Lehmann, Christian H. K.; Hatscher, Lukas; Purbojo, Ariawan; Hartmann, Arndt; Garcia-Martin, Fayna; Nishimura, Shin-Ichiro; Cesnjevar, Robert; Nimmerjahn, Falk; Dudziak, Diana
    Frontiers in Immunology (2018), 9 (), 744/1-744/16CODEN: FIRMCW; ISSN:1664-3224. (Frontiers Media S.A.)
    Dendritic cells (DCs) are major players for the induction of immune responses. Apart from plasmacytoid DCs (pDCs), human DCs can be categorized into two types of conven-tional DCs: CD141+ DCs (cDC1) and CD1c+ DCs (cDC2). Defining uniquely expressed surface markers on human immune cells is not only important for the identification of DC subpopulations but also a prerequisite for harnessing the DC subset-specific potential in immunomodulatory approaches, such as antibody-mediated antigen targeting. Although others identified CLEC9A as a specific endocytic receptor for CD141+ DCs, such a recep-tor for CD1c+ DCs has not been discovered, yet. By performing transcriptomic and flow cytometric analyses on human DC subpopulations from different lymphohematopoietic tissues, we identified CLEC10A (CD301, macrophage galactose-type C-type lectin) as a specific marker for human CD1c+ DCs. We further demonstrate that CLEC10A rapidly internalizes into human CD1c+ DCs upon binding of a monoclonal antibody directed against CLEC10A. The binding of a CLEC10A-specific bivalent ligand (the MUC-1 pep-tide glycosylated with N-acetylgalactosamine) is limited to CD1c+ DCs and enhances the cytokine secretion (namely TNFα, IL-8, and IL-10) induced by TLR 7/8 stimulation. Thus, CLEC10A represents not only a candidate to better define CD1c+ DCs-due to its high endocytic potential-CLEC10A also exhibits an interesting candidate receptor for future antigen-targeting approaches.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXit1Gls7vK&md5=2f4e523c6fac15e319656205747b0255
  27. 27
    Napoletano, C.; Rughetti, A.; Agervig Tarp, M. P.; Coleman, J.; Bennett, E. P.; Picco, G.; Sale, P.; Denda-Nagai, K.; Irimura, T.; Mandel, U.; Clausen, H.; Frati, L.; Taylor-Papadimitriou, J.; Burchell, J.; Nuti, M. Tumor-Associated Tn-MUC1 Glycoform Is Internalized through the Macrophage Galactose-Type C-Type Lectin and Delivered to the HLA Class I and II Compartments in Dendritic Cells. Cancer Res. 2007, 67, 8358– 8367,  DOI: 10.1158/0008-5472.can-07-1035
    Google Scholar
    27
    Tumor-Associated Tn-MUC1 Glycoform Is Internalized through the Macrophage Galactose-Type C-Type Lectin and Delivered to the HLA Class I and II Compartments in Dendritic Cells
    Napoletano, Chiara; Rughetti, Aurelia; Agervig Tarp, Mads P.; Coleman, Julia; Bennett, Eric P.; Picco, Gianfranco; Sale, Patrizio; Denda-Nagai, Kaori; Irimura, Tatsuro; Mandel, Ulla; Clausen, Henrik; Frati, Luigi; Taylor-Papadimitriou, Joyce; Burchell, Joy; Nuti, Marianna
    Cancer Research (2007), 67 (17), 8358-8367CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
    The type of interaction between tumor-assocd. antigens and specialized antigen-presenting cells such as dendritic cells (DCs) is crit. for the type of immunity that will be generated. MUC1, a highly O-glycosylated mucin, is overexpressed and aberrantly glycosylated in several tumor histiotypes. This results in the expression of tumor-assocd. glycoforms and in MUC1 carrying the tumor-specific glycan Tn (GalNAcα1-O-Ser/Thr). Glycopeptides corresponding to three tandem repeats of MUC1, enzymically glycosylated with 9 or 15 mol of GalNAc, were shown to specifically bind and to be internalized by immature monocyte-derived DCs (iDCs). Binding required calcium and the GalNAc residue and was competed out by GalNAc polymer and Tn-MUC1 or Tn-MUC2 glycopeptides. The macrophage galactose-type C-type lectin (MGL) receptor expressed on iDCs was shown to be responsible for the binding. Confocal anal. and ELISA done on subcellular fractions of iDCs showed that the Tn-MUC1 glycopeptides colocalized with HLA class I and II compartments after internalization. Importantly, although Tn-MUC1 recombinant protein was bound and internalized by MGL, the glycoprotein entered the HLA class II compartment, but not the HLA class I pathway. These data indicate that MGL expressed on iDCs is an optimal receptor for the internalization of short GalNAcs carrying immunogens to be delivered into HLA class I and II compartments. Such glycopeptides therefore represent a new way of targeting the HLA class I and II pathways of DCs. These results have possible implications in designing cancer vaccines.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXpvFKisr4%253D&md5=d1d5372f51e7a10194abd3254298342b
  28. 28
    Napoletano, C.; Zizzari, I. G.; Rughetti, A.; Rahimi, H.; Irimura, T.; Clausen, H.; Wandall, H. H.; Belleudi, F.; Bellati, F.; Pierelli, L.; Frati, L.; Nuti, M. Targeting of Macrophage Galactose-Type C-Type Lectin (MGL) Induces DC Signaling and Activation. Eur. J. Immunol. 2012, 42, 936– 945,  DOI: 10.1002/eji.201142086
    Google Scholar
    28
    Targeting of macrophage galactose-type C-type lectin (MGL) induces DC signaling and activation
    Napoletano, Chiara; Zizzari, Ilaria G.; Rughetti, Aurelia; Rahimi, Hassan; Irimura, Tatsuro; Clausen, Henrik; Wandall, Hans H.; Belleudi, Francesca; Bellati, Filippo; Pierelli, Luca; Frati, Luigi; Nuti, Marianna
    European Journal of Immunology (2012), 42 (4), 936-945CODEN: EJIMAF; ISSN:0014-2980. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Dendritic cells (DCs) sense the microenvironment through several types of receptors recognizing pathogen-assocd. mol. patterns. In particular, C-type lectins, expressed by distinct subsets of DCs, recognize and internalize specific carbohydrate antigen in a Ca2+-dependent manner. Targeting of these receptors is becoming an efficient strategy of delivering antigens in DC-based anticancer immunotherapy. Here we investigated the role of the macrophage galactose type C-lectin receptor (MGL), expressed by immature DCs (iDCs), as a mol. target for α-N-acetylgalactosamine (GalNAc or Tn)-carrying tumor-assocd. antigens to improve DC performance. MGL expressed by ex vivo-generated iDCs from healthy donors was engaged by a 60-mer MUC19Tn-glycopeptide as a Tn-carrying tumor-assocd. antigen, and an anti-MGL antibody, as a specific MGL binder. We demonstrated that MGL engagement induced homotrimers and homodimers, triggering the phosphorylation of extracellular signal-regulated kinase 1,2 (ERK1,2) and nuclear factor-κB activation. Anal. of DC phenotype and function demonstrated that MGL engagement improved DC performance as antigen-presenting cells, promoting the upregulation of maturation markers, a decrease in phagocytosis, an enhancement of motility, and most importantly an increase in antigen-specific CD8+T-cell activation. These results demonstrate that the targeting of MGL receptor on human DCs has an adjuvant effect and that this strategy can be used to design novel anticancer vaccines.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XmtVKktr0%253D&md5=1937d82644d4875e83734ad6623e849f
  29. 29
    Gabba, A.; Bogucka, A.; Luz, J. G.; Diniz, A.; Coelho, H.; Corzana, F.; Cañada, F. J.; Marcelo, F.; Murphy, P. V.; Birrane, G. Crystal Structure of the Carbohydrate Recognition Domain of the Human Macrophage Galactose C-Type Lectin Bound to GalNAc and the Tumor-Associated Tn Antigen. Biochemistry 2021, 60, 1327– 1336,  DOI: 10.1021/acs.biochem.1c00009
    Google Scholar
    29
    Crystal Structure of the Carbohydrate Recognition Domain of the Human Macrophage Galactose C-Type Lectin Bound to GalNAc and the Tumor-Associated Tn Antigen
    Gabba, Adele; Bogucka, Agnieszka; Luz, John G.; Diniz, Ana; Coelho, Helena; Corzana, Francisco; Canada, Francisco Javier; Marcelo, Filipa; Murphy, Paul V.; Birrane, Gabriel
    Biochemistry (2021), 60 (17), 1327-1336CODEN: BICHAW; ISSN:0006-2960. (American Chemical Society)
    The human macrophage galactose lectin (MGL) is an endocytic type ii transmembrane receptor expressed on immature monocyte-derived dendritic cells and activated macrophages and plays a role in modulating the immune system in response to infections and cancer. MGL contains an extracellular calcium-dependent (C-type) carbohydrate recognition domain (CRD) that specifically binds terminal N-acetylgalactosamine glycan residues such as the Tn and sialyl-Tn antigens found on tumor cells, as well as other N- and O-glycans displayed on certain viruses and parasites. Even though the glycan specificity of MGL is known and several binding glycoproteins were identified, the mol. basis for substrate recognition has remained elusive due to the lack of high-resoln. structures. Here the authors present crystal structures of the MGL CRD at near endosomal pH and in several complexes, which reveal details of the interactions with the natural ligand, GalNAc, the cancer-assocd. Tn-Ser antigen, and a synthetic GalNAc mimetic ligand. Like the asialoglycoprotein receptor, addnl. calcium atoms are present and contribute to stabilization of the MGL CRD fold. The structure provides the mol. basis for preferential binding of N-acetylgalactosamine over galactose and prompted the reevaluation of the binding modes previously proposed in soln. Satn. transfer difference NMR data acquired using the MGL CRD and interpreted using the crystal structure indicate a single binding mode for GalNAc in soln. Models of MGL1 and MGL2, the mouse homologs of MGL, explain how these proteins might recognize LewisX and GalNAc, resp.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXmsVWjt74%253D&md5=bfc979febdd29a4b74685f1439c2274a
  30. 30
    André, S.; O’Sullivan, S.; Koller, C.; Murphy, P. V.; Gabius, H. J. Bi- to Tetravalent Glycoclusters Presenting GlcNAc/GalNAc as Inhibitors: From Plant Agglutinins to Human Macrophage Galactose-Type Lectin (CD301) and Galectins. Org. Biomol. Chem. 2015, 13, 4190– 4203,  DOI: 10.1039/c5ob00048c
    Google Scholar
    30
    Bi- to tetravalent glycoclusters presenting GlcNAc/GalNAc as inhibitors: from plant agglutinins to human macrophage galactose-type lectin (CD301) and galectins
    Andre, Sabine; O'Sullivan, Shane; Koller, Christiane; Murphy, Paul V.; Gabius, Hans-Joachim
    Organic & Biomolecular Chemistry (2015), 13 (14), 4190-4203CODEN: OBCRAK; ISSN:1477-0520. (Royal Society of Chemistry)
    Emerging insights into the functional spectrum of tissue lectins leads to identification of new targets for the custom-made design of potent inhibitors, providing a challenge for synthetic chem. The affinity and selectivity of a carbohydrate ligand for a lectin may immensely be increased by a no. of approaches, which includes varying geometrical or topol. features. This perspective leads to the design and synthesis of glycoclusters and their testing using assays of physiol. relevance. Herein, hydroquinone, resorcinol, benzene-1,3,5-triol and tetra(4-hydroxyphenyl)ethene have been employed as scaffolds and propargyl derivs. obtained. The triazole-contg. linker to the α/β-O/S-glycosides of GlcNAc/GalNAc presented on these scaffolds was generated by copper-catalyzed azide-alkyne cycloaddn. This strategy was used to give a panel of nine glycoclusters with bi-, tri- and tetravalency. Maintained activity for lectin binding after conjugation was ascertained for both sugars in solid-phase assays with the plant agglutinins WGA (GlcNAc) and DBA (GalNAc). Absence of cross-reactivity excluded any carbohydrate-independent reactivity of the bivalent compds., allowing the authors to proceed to further testing with a biomedically relevant lectin specific for GalNAc. Macrophage galactose(-binding C)-type lectin, involved in immune defense by dendritic cells and in virus uptake, was produced as a sol. protein without/with its α-helical coiled-coil stalk region. Binding to ligands presented on a matrix and on cell surfaces was highly susceptible to the presence of the tetravalent inhibitor derived from the tetraphenylethene-contg. scaffold, and presentation of GalNAc with an α-thioglycosidic linkage proved favorable. Cross-reactivity of this glycocluster to human galectins-3 and -4, which interact with Tn-antigen-presenting mucins, was rather small. Evidently, the valency and spatial display of α-GalNAc residues is a key factor to design potent and selective inhibitors for this lectin.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXjtVemtbw%253D&md5=0d13d8cd20d4c914636876b34ecee15a
  31. 31
    Kaltner, H.; Manning, J. C.; García Caballero, G.; Di Salvo, C.; Gabba, A.; Romero-Hernández, L. L.; Knospe, C.; Wu, D.; Daly, H. C.; O’Shea, D. F.; Gabius, H. J.; Murphy, P. V. Revealing Biomedically Relevant Cell and Lectin Type-Dependent Structure-Activity Profiles for Glycoclusters by Using Tissue Sections as an Assay Platform. RSC Adv. 2018, 8, 28716– 28735,  DOI: 10.1039/c8ra05382k
    Google Scholar
    31
    Revealing biomedically relevant cell and lectin type-dependent structure-activity profiles for glycoclusters by using tissue sections as an assay platform
    Kaltner, Herbert; Manning, Joachim C.; Garcia Caballero, Gabriel; Di Salvo, Claudia; Gabba, Adele; Romero-Hernandez, Laura L.; Knospe, Clemens; Wu, Dan; Daly, Harrison C.; O'Shea, Donal F.; Gabius, Hans-Joachim; Murphy, Paul V.
    RSC Advances (2018), 8 (50), 28716-28735CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)
    The increasing realization of the involvement of lectin-glycan recognition in (patho)physiol. processes inspires envisioning therapeutic intervention by high-avidity/specificity blocking reagents. Synthetic glycoclusters are proving to have potential for becoming such inhibitors but the commonly used assays have their drawbacks to predict in vivo efficacy. They do not represent the natural complexity of (i) cell types and (ii) spatial and structural complexity of glycoconjugate representation. Moreover, testing lectins in mixts., as present in situ, remains a major challenge, giving direction to this work. Using a toolbox with four lectins and six bi- to tetravalent glycoclusters bearing the cognate sugar in a model study, we here document the efficient and versatile application of tissue sections (from murine jejunum as the model) as a platform for routine and systematic glycocluster testing without commonly encountered limitations. The nature of glycocluster structure, esp. core and valency, and of protein features, i.e. architecture, fine-specificity and valency, are shown to have an influence, as cell types can differ in response profiles. Proceeding from light microscopy to monitoring by fluorescence microscopy enables grading of glycocluster activity on individual lectins tested in mixts. This work provides a robust tool for testing glycoclusters prior to considering in vivo expts.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhsV2itr3M&md5=54f546a60e3039cd0aeb157e0210375c
  32. 32
    Tseng, N. W.; Liu, J.; Ng, J. C. Y.; Lam, J. W. Y.; Sung, H. H. Y.; Williams, I. D.; Tang, B. Z. Deciphering Mechanism of Aggregation-Induced Emission (AIE): Is E-Z Isomerisation Involved in an AIE Process?. Chem. Sci. 2012, 3, 493– 497,  DOI: 10.1039/c1sc00690h
    Google Scholar
    32
    Deciphering mechanism of aggregation-induced emission (AIE): Is E-Z isomerisation involved in an AIE process?
    Tseng, Nai-Wen; Liu, Jianzhao; Ng, Jason C. Y.; Lam, Jacky W. Y.; Sung, Herman H. Y.; Williams, Ian D.; Tang, Ben Zhong
    Chemical Science (2012), 3 (2), 493-497CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
    In this work, we address a mechanistic issue on AIE process and correct a long-held misconception on stilbene photoluminescence. E-Z isomerization has been generally recognized as the cause of emission quenching in stilbene solns. A natural question arisen from this common belief is whether suppression of E-Z isomerization by aggregate formation in a stilbenic fluorogen system is responsible for its AIE phenomenon. Monitoring of the structural change of a stilbene deriv. named 1,2-diphenyl-1,2-di(p-tolyl)ethene by NMR during UV irradn. reveals that the E-Z isomerization is not involved in its AIE process under the normal photoluminescence spectral measurement conditions.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XkslaqsQ%253D%253D&md5=e978caacf20d1ce780445f43518662cc
  33. 33
    Donnier-Maréchal, M.; Abdullayev, S.; Bauduin, M.; Pascal, Y.; Fu, M.-Q.; He, X.-P.; Gillon, E.; Imberty, A.; Kipnis, E.; Dessein, R.; Vidal, S. Tetraphenylethylene-Based Glycoclusters with Aggregation-Induced Emission (AIE) Properties as High-Affinity Ligands of Bacterial Lectins. Org. Biomol. Chem. 2018, 16, 8804– 8809,  DOI: 10.1039/c8ob02035c
    Google Scholar
    33
    Tetraphenylethylene-based glycoclusters with aggregation-induced emission (AIE) properties as high-affinity ligands of bacterial lectins
    Donnier-Marechal, Marion; Abdullayev, Shuay; Bauduin, Marvin; Pascal, Yoann; Fu, Meng-Qi; He, Xiao-Peng; Gillon, Emilie; Imberty, Anne; Kipnis, Eric; Dessein, Rodrigue; Vidal, Sebastien
    Organic & Biomolecular Chemistry (2018), 16 (45), 8804-8809CODEN: OBCRAK; ISSN:1477-0520. (Royal Society of Chemistry)
    Tetraphenylethylene (TPE) is fluorescent through aggregation induced emission (AIE) in water. Herein, TPE was used as the core of glycoclusters that target the bacterial lectins LecA and LecB of Pseudomonas aeruginosa. Synthesis of these TPE-based glycoclusters was accomplished by using azide-alkyne "click" chem. The AIE properties of the resulting glycoclusters could be readily verified, but imaging could not be pursued due to the overlap of the fluorescence signals from cells and bacteria. Nonetheless, the glycoclusters displayed nanomolar affinities toward LecA and LecB. Further evaluation in a cell-based anti-adhesive assay highlighted a limited decrease in adhesion (20%) for the fucosylated glycocluster. This confirmed that these TPE-based glycoclusters are indeed LecA and LecB high-affinity ligands. Nevertheless, the hypotheses involving their application in imaging or anti-adhesive therapy could not be verified.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhvFyntLzJ&md5=df9ac1602341695242757dbd943ff469
  34. 34
    Jégouzo, S. A.; Quintero-Martínez, A.; Ouyang, X.; Dos Santos, Á.; Taylor, M. E.; Drickamer, K. Organization of the Extracellular Portion of the Macrophage Galactose Receptor: A Trimeric Cluster of Simple Binding Sites for N-Acetylgalactosamine. Glycobiology 2013, 23, 853– 864,  DOI: 10.1093/glycob/cwt022
    Google Scholar
    34
    Organization of the extracellular portion of the macrophage galactose receptor: A trimeric cluster of simple binding sites for N-acetylgalactosamine
    Jegouzo, Sabine A. F.; Quintero-Martinez, Adrian; Ouyang, Xiangyu; dos Santos, Alia; Taylor, Maureen E.; Drickamer, Kurt
    Glycobiology (2013), 23 (7), 853-864CODEN: GLYCE3; ISSN:0959-6658. (Oxford University Press)
    The properties of the human macrophage galactose receptor have been investigated. Specificity for N-acetylgalactosamine (GalNAc) residues with exposed 3- and 4-hydroxyl groups explains virtually all of the results obtained from a recently expanded array of synthetic glycans and is consistent with a model for the structure of the binding site. This simple interaction is sufficient to explain the ability of the receptor to bind to tumor-cell glycans bearing Tn and sialyl-Tn antigens, but not to more elaborate O-linked glycans that predominate on normal cells. This specificity also allows for binding of parasite glycans and screening of an array of bacterial outer membrane oligosaccharides confirms that the receptor binds to a subset of these structures with appropriately exposed GalNAc residues. A key feature of the receptor is the clustering of binding sites in the extracellular portion of the protein, which retains the trimeric structure obsd. in the cell membrane. Chem. crosslinking, gel filtration, CD anal. and differential scanning calorimetry demonstrate that this trimeric structure of the receptor is stabilized by an α-helical coiled coil that extends from the surface of the membrane to the globular carbohydrate-recognition domains. The helical neck domains form independent trimerization domains. Taken together, these results indicate that the macrophage galactose receptor shares many of the features of serum mannose-binding protein, in which clusters of monosaccharide-binding sites serve as detectors for a simple epitope that is not common on endogenous cell surface glycans but that is abundant on the surfaces of tumor cells and certain pathogens.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXptFerurk%253D&md5=dac97ed7fb3dbee78fffe466d1314cfe
  35. 35
    Hu, X. M.; Chen, Q.; Wang, J. X.; Cheng, Q. Y.; Yan, C. G.; Cao, J.; He, Y. J.; Han, B. H. Tetraphenylethylene-Based Glycoconjugate as a Fluorescence “Turn-on” Sensor for Cholera Toxin. Chem.─Asian J. 2011, 6, 2376– 2381,  DOI: 10.1002/asia.201100141
    Google Scholar
    35
    Tetraphenylethylene-based Glycoconjugate as a Fluorescence "Turn-On" Sensor for Cholera Toxin
    Hu, Xin-Ming; Chen, Qi; Wang, Jin-Xiang; Cheng, Qian-Yi; Yan, Chao-Guo; Cao, Jie; He, Yu-Jian; Han, Bao-Hang
    Chemistry - An Asian Journal (2011), 6 (9), 2376-2381CODEN: CAAJBI; ISSN:1861-4728. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Tetraphenylethylene (TPE)-based glycoconjugates were easily synthesized by copper(I)-catalyzed "click reactions" between propargyl-attached TPE and azido-functionalized sugars. The TPE compd. bearing lactosyl moieties (Lac-TPE) was found to be a fluorescence "turn-on" sensor for cholera toxin by virtue of aggregation-induced emission characteristics of the TPE motif owing to the specific interaction of lactose with the cholera toxin B subunit, while a cellobiose-functionalized TPE deriv. did not show any response to the toxin. Therefore, Lac-TPE shows promising applications in the detection of cholera toxin, as well as in the investigation of carbohydrate-protein interaction.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXoslCrsrY%253D&md5=af3f2d2fede95ef75314655ded79cb5f
  36. 36
    Price, M. R.; Rye, P. D.; Petrakou, E.; Murray, A.; Brady, K.; Imai, S.; Haga, S.; Kiyozuka, Y.; Schol, D.; Meulenbroek, M. F. A.; Snijdewint, F. G. M.; Von Mensdorff-Pouilly, S.; Verstraeten, R. A.; Nil, K.; Blockzjil, A.; Nil, N.; Nilsson, O.; Nil, R.; Suresh, M. R.; Nil, K.; Fortier, S.; Nil, B.; Berg, A.; Longenecker, M. B.; Nil, H.; Boer, M.; Nil, K.; McKenzie, I. F. C.; Nil, G.; Simeoni, L. A.; Ter-Grigoryan, A. G.; Belyanchikov, I. M.; Bovin, N. V.; Cao, Y.; Karsten, U.; Dai, J.; Allard, W. J.; Davis, G.; Yeung, K. K.; Hanisch, F. G.; Lloyd, K. O.; Kudryashov, V.; Sikut, R.; Sikut, A.; Zhang, K.; Baeckström, D.; Hansson, G. C.; Reis, C. A.; Hassan, H.; Bennett, E. P.; Claussen, H.; Norum, L.; Varaas, T.; Kierulf, B.; Nustad, K.; Ciborowski, P.; Konitzki, W. M.; Magarian-Blander, J.; Finn, O. J.; Hilgers, J. Summary Report on the ISOBM TD-4 Workshop: Analysis of 56 Monoclonal Antibodies against the MUC1 Mucin. Tumor Biol. 1998, 19, 1– 20,  DOI: 10.1159/000056500
    Google Scholar
    36
    Summary report on the ISOBM TD-4 Workshop: analysis of 56 monoclonal antibodies against the MUC1 mucin. San Diego, Calif., November 17-23, 1996
    Price M R; Rye P D; Petrakou E; Murray A; Brady K; Imai S; Haga S; Kiyozuka Y; Schol D; Meulenbroek M F; Snijdewint F G; von Mensdorff-Pouilly S; Verstraeten R A; Kenemans P; Blockzjil A; Nilsson K; Nilsson O; Reddish M; Suresh M R; Koganty R R; Fortier S; Baronic L; Berg A; Longenecker M B; Hilgers J; et al
    Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine (1998), 19 Suppl 1 (), 1-20 ISSN:1010-4283.
    Sixteen research groups participated in the ISOBM TD-4 Workshop in which the reactivity and specificity of 56 monoclonal antibodies against the MUC1 mucin was investigated using a diverse panel of target antigens and MUC1 mucin-related synthetic peptides and glycopeptides. The majority of antibodies (34/56) defined epitopes located within the 20-amino acid tandem repeat sequence of the MUC1 mucin protein core. Of the remaining 22 antibodies, there was evidence for the involvement of carbohydrate residues in the epitopes for 16 antibodies. There was no obvious relationship between the type of immunogen and the specificity of each antibody. Synthetic peptides and glycopeptides were analyzed for their reactivity with each antibody either by assay of direct binding (e.g. by ELISA or BiaCore) or by determining the capacity of synthetic ligands to inhibit antibody binding interactions. There was good concordance between the research groups in identifying antibodies reactive with peptide epitopes within the MUC1 protein core. Epitope mapping tests were performed using the Pepscan analysis for antibody reactivity against overlapping synthetic peptides, and results were largely consistent between research groups. The dominant feature of epitopes within the MUC1 protein core was the presence, in full or part, of the hydrophilic sequence of PDTRAPAP. Carbohydrate epitopes were less easily characterized and the most useful reagents in this respect were defined oligosaccharides, rather than purified mucin preparations enriched in particular carbohydrate moieties. It was evident that carbohydrate residues were involved in many epitopes, by regulating epitope accessibility or masking determinants, or by stabilizing preferred conformations of peptide epitopes within the MUC1 protein core. Overall, the studies, highlight concordance between groups rather than exposing inconsistencies which gives added confidence to the results of analyses of the specificity of antimucin monoclonal antibodies.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADyaK1c%252FosVSjsA%253D%253D&md5=18abcd625acb7d3e72598493a1abc31b
  37. 37
    Pett, C.; Cai, H.; Liu, J.; Palitzsch, B.; Schorlemer, M.; Hartmann, S.; Stergiou, N.; Lu, M.; Kunz, H.; Schmitt, E.; Westerlind, U. Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. Chem.─Eur. J. 2017, 23, 3875– 3884,  DOI: 10.1002/chem.201603921
    Google Scholar
    37
    Microarray analysis of antibodies induced with synthetic antitumor vaccines: Specificity against diverse mucin core structures
    Pett, Christian; Cai, Hui; Liu, Jia; Palitzsch, Bjoern; Schorlemer, Manuel; Hartmann, Sebastian; Stergiou, Natascha; Lu, Mengji; Kunz, Horst; Schmitt, Edgar; Westerlind, Ulrika
    Chemistry - A European Journal (2017), 23 (16), 3875-3884CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Glycoprotein research is pivotal for vaccine development and biomarker discovery. Many successful methodologies for reliably increasing the antigenicity toward tumor-assocd. glycopeptide structures have been reported. Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunol. memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunol. For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-assocd. glycans (TN, STN, and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable d. at the five different sites of the MUC1 tandem repeat. This is one of the most extensive glycopeptide libraries ever made through total synthesis. On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amts. of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-assocd. antigen structures. The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. It is obsd. that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. All antibody sera show high reactivity to the tumor-assocd. saccharide structures on MUC1. Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhsVaqs7c%253D&md5=6af0f627abe8d7c19adef5f7c7cea23a
  38. 38
    Wandall, H. H.; Blixt, O.; Tarp, M. A.; Pedersen, J. W.; Bennett, E. P.; Mandel, U.; Ragupathi, G.; Livingston, P. O.; Hollingsworth, M. A.; Taylor-Papadimitriou, J.; Burchell, J.; Clausen, H. Cancer Biomarkers Defined by Autoantibody Signatures to Aberrant O-Glycopeptide Epitopes. Cancer Res. 2010, 70, 1306– 1313,  DOI: 10.1158/0008-5472.can-09-2893
    Google Scholar
    38
    Cancer Biomarkers Defined by Autoantibody Signatures to Aberrant O-Glycopeptide Epitopes
    Wandall, Hans H.; Blixt, Ola; Tarp, Mads A.; Pedersen, Johannes W.; Bennett, Eric P.; Mandel, Ulla; Ragupathi, Govind; Livingston, Phil O.; Hollingsworth, Michael A.; Taylor-Papadimitriou, Joyce; Burchell, Joy; Clausen, Henrik
    Cancer Research (2010), 70 (4), 1306-1313CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
    Autoantibodies to cancer antigens hold promise as biomarkers for early detection of cancer. Proteins that are aberrantly processed in cancer cells are likely to present autoantibody targets. The extracellular mucin MUC1 is overexpressed and aberrantly glycosylated in many cancers; thus, the authors evaluated whether autoantibodies generated to aberrant O-glycoforms of MUC1 might serve as sensitive diagnostic biomarkers for cancer. Using an antibody-based glycoprofiling ELISA assay, the authors documented that aberrant truncated glycoforms were not detected in sera of cancer patients. An O-glycopeptide microarray was developed that detected IgG antibodies to aberrant O-glycopeptide epitopes in patients vaccinated with a keyhole limpet hemocyanin-conjugated truncated MUC1 peptide. The authors detected cancer-assocd. IgG autoantibodies in sera from breast, ovarian, and prostate cancer patients against different aberrent O-glycopeptide epitopes derived from MUC1. These autoantibodies represent a previously unaddressed source of sensitive biomarkers for early detection of cancer. The methods the authors have developed for chemoenzymic synthesis of O-glycopeptides on microarrays may allow for broader mining of the entire cancer O-glycopeptidome. Cancer Res; 70(4); 1306-13.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhvFaksrc%253D&md5=f21b0b73d10e93808720c84f4f4c8711
  39. 39
    Borgert, A.; Heimburg-Molinaro, J.; Song, X.; Lasanajak, Y.; Ju, T.; Liu, M.; Thompson, P.; Ragupathi, G.; Barany, G.; Smith, D. F.; Cummings, R. D.; Live, D. Deciphering Structural Elements of Mucin Glycoprotein Recognition. ACS Chem. Biol. 2012, 7, 1031– 1039,  DOI: 10.1021/cb300076s
    Google Scholar
    39
    Deciphering Structural Elements of Mucin Glycoprotein Recognition
    Borgert, Andrew; Heimburg-Molinaro, Jamie; Song, Xuezheng; Lasanajak, Yi; Ju, Tongzhong; Liu, Mian; Thompson, Pamela; Ragupathi, Govind; Barany, George; Smith, David F.; Cummings, Richard D.; Live, David
    ACS Chemical Biology (2012), 7 (6), 1031-1039CODEN: ACBCCT; ISSN:1554-8929. (American Chemical Society)
    Mucin glycoproteins present a complex structural landscape arising from the multiplicity of glycosylation patterns afforded by their numerous serine and threonine glycosylation sites, often in clusters, and with variations in resp. glycans. To explore the structural complexities in such glycoconjugates, the authors used NMR to systematically analyze the conformational effects of glycosylation d. within a cluster of sites. This allows correlation with mol. recognition through anal. of interactions between these and other glycopeptides, with antibodies, lectins, and sera, using a glycopeptide microarray. Selective antibody interactions with discrete conformational elements, reflecting aspects of the peptide and disposition of GalNAc residues, are obsd. The authors' results help bridge the gap between conformational properties and mol. recognition of these mols., with implications for their physiol. roles. Features of the native mucin motifs impact their relative immunogenicity and are accurately encoded in the antibody binding site, with the conformational integrity being preserved in isolated glycopeptides, as reflected in the antibody binding profile to array components.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XksVeksrw%253D&md5=c37329341e9dd4ef7681a22906532ea1
  40. 40
    Coltart, D. M.; Royyuru, A. K.; Williams, L. J.; Glunz, P. W.; Sames, D.; Kuduk, S. D.; Schwarz, J. B.; Chen, X. T.; Danishefsky, S. J.; Live, D. H. Principles of Mucin Architecture: Structural Studies on Synthetic Glycopeptides Bearing Clustered Mono-Di-Tri-and Hexasaccharide Glycodomains. J. Am. Chem. Soc. 2002, 124, 9833– 9844,  DOI: 10.1021/ja020208f
    Google Scholar
    40
    Principles of Mucin Architecture: Structural Studies on Synthetic Glycopeptides Bearing Clustered Mono-, Di-, Tri-, and Hexasaccharide Glycodomains
    Coltart, Don M.; Royyuru, Ajay K.; Williams, Lawrence J.; Glunz, Peter W.; Sames, Dalibor; Kuduk, Scott D.; Schwarz, Jacob B.; Chen, Xiao-Tao; Danishefsky, Samuel J.; Live, David H.
    Journal of the American Chemical Society (2002), 124 (33), 9833-9844CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    The structural characteristics of a mucin glycopeptide motif derived from the N-terminal fragment STTAV of the cell surface glycoprotein CD43 have been investigated by NMR. In this study, a series of mols. prepd. by total synthesis were examd., consisting of the peptide itself, three glycopeptides having clustered sites of α-O-glycosylation on the serine and threonine side chains with the Tn, TF, and STF carbohydrate antigens, resp., and one with the β-O-linked TF antigen. Addnl., a glycopeptide having the sequence SSSAVAV, triglycosylated with the Ley epitope, was investigated. NMR data for the tri-STF-STTAV glycopeptide were used to solve the structure of this construct through restrained mol. dynamics calcns. The calcns. revealed a defined conformation for the glycopeptide core rooted in the interaction of the peptide and the first N-acetylgalactosamine residue. The similarity of the NMR data for each of the α-O-linked glycopeptides demonstrates that this structure persists for each construct and that the mode of attachment of the first sugar and the peptide is paramount in establishing the organization of the core. The core provides a common framework on which a variety of glycans may be displayed. Remarkably, while there is a profound organizational effect on the peptide backbone with the α-linked glycans, attachment via a β-linkage has little apparent consequence.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XlsFygtL8%253D&md5=db84e77f38b2cba8d3645af74b80257f
  41. 41
    Apostolopoulos, V.; Yuriev, E.; Ramsland, P. A.; Halton, J.; Osinski, C.; Li, W.; Plebanski, M.; Paulsen, H.; McKenzie, I. F. C. A Glycopeptide in Complex with MHC Class I Uses the GalNAc Residue as an Anchor. Proc. Natl. Acad. Sci. U.S.A. 2003, 100, 15029– 15034,  DOI: 10.1073/pnas.2432220100
    Google Scholar
    41
    A glycopeptide in complex with MHC class I uses the GalNAc residue as an anchor
    Apostolopoulos, Vasso; Yuriev, Elizabeth; Ramsland, Paul A.; Halton, Jodie; Osinski, Carla; Li, Wenjun; Plebanski, Magdalena; Paulsen, Hans; McKenzie, Ian F. C.
    Proceedings of the National Academy of Sciences of the United States of America (2003), 100 (25), 15029-15034CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
    Peptides bind MHC class I mols. by anchoring hydrophobic side chains into pockets in the peptide binding groove. Here, we report an immunogenic (in vitro and in vivo) MUC1 glycopeptide (MUC1-8-5GalNAc) bound to H-2Kb, fully crossreactive with the nonglycosylated variant. Mol. modeling showed that the central P5-Thr-GalNAc residue points into the C pocket and forms van der Waals and hydrogen bond interactions with the MHC class I. As predicted, GalNAc, a modified peptide carrying an addnl. anchor in the central C anchor pocket, increased the affinity by ≈100-fold compared with the native low-affinity peptide (MUC1-8). The findings demonstrate that glycopeptides assocd. with MHC class I mols. can use GalNAc to anchor the peptide in the groove and enable high-affinity binding.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXpvFaqtLY%253D&md5=22feb349b128c699a317d5d103262948
  42. 42
    Avci, F. Y.; Li, X.; Tsuji, M.; Kasper, D. L. A Mechanism for Glycoconjugate Vaccine Activation of the Adaptive Immune System and Its Implications for Vaccine Design. Nat. Med. 2011, 17, 1602– 1609,  DOI: 10.1038/nm.2535
    Google Scholar
    42
    A mechanism for glycoconjugate vaccine activation of the adaptive immune system and its implications for vaccine design
    Avci, Fikri Y.; Li, Xiangming; Tsuji, Moriya; Kasper, Dennis L.
    Nature Medicine (New York, NY, United States) (2011), 17 (12), 1602-1609CODEN: NAMEFI; ISSN:1078-8956. (Nature Publishing Group)
    Glycoconjugate vaccines have provided enormous health benefits globally, but they have been less successful in some populations at high risk for developing disease. To identify new approaches to enhancing glycoconjugate effectiveness, we investigated mol. and cellular mechanisms governing the immune response to a prototypical glycoconjugate vaccine. We found that in antigen-presenting cells a carbohydrate epitope is generated upon endolysosomal processing of group B streptococcal type III polysaccharide coupled to a carrier protein. In conjunction with a carrier protein-derived peptide, this carbohydrate epitope binds major histocompatibility class II (MHCII) and stimulates carbohydrate-specific CD4+ T cell clones to produce interleukins 2 and 4-cytokines essential for providing T cell help to antibody-producing B cells. An archetypical glycoconjugate vaccine that we constructed to maximize the presentation of carbohydrate-specific T cell epitopes is 50-100 times more potent and substantially more protective in a neonatal mouse model of group B Streptococcus infection than a vaccine constructed by methods currently used by the vaccine industry. Our discovery of how glycoconjugates are processed resulting in presentation of carbohydrate epitopes that stimulate CD4+ T cells has key implications for glycoconjugate vaccine design that could result in greatly enhanced vaccine efficacy.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsV2gt7%252FL&md5=c02d36037865b38be13179ca1eb84fce
  43. 43
    Vinuesa, C. G.; Linterman, M. A.; Yu, D.; Maclennan, I. C. M. Follicular Helper T Cells. Annu. Rev. Immunol. 2016, 34, 335– 368,  DOI: 10.1146/annurev-immunol-041015-055605
    Google Scholar
    43
    Follicular Helper T Cells
    Vinuesa, Carola G.; Linterman, Michelle A.; Yu, Di; MacLennan, Ian C. M.
    Annual Review of Immunology (2016), 34 (), 335-368CODEN: ARIMDU; ISSN:0732-0582. (Annual Reviews)
    Although T cell help for B cells was described several decades ago, it was the identification of CXCR5 expression by B follicular helper T (Tfh) cells and the subsequent discovery of their dependence on BCL6 that led to the recognition of Tfh cells as an independent helper subset and accelerated the pace of discovery. More than 20 transcription factors, together with RNA-binding proteins and microRNAs, control the expression of chemotactic receptors and mols. important for the function and homeostasis of Tfh cells. Tfh cells prime B cells to initiate extrafollicular and germinal center antibody responses and are crucial for affinity maturation and maintenance of humoral memory. In addn. to the roles that Tfh cells have in antimicrobial defense, in cancer, and as HIV reservoirs, regulation of these cells is crit. to prevent autoimmunity. The realization that follicular T cells are heterogeneous, comprising helper and regulatory subsets, has raised questions regarding a possible division of labor in germinal center B cell selection and elimination.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XivFKlsrw%253D&md5=4c8dc98065906b9115c3876d236f16be
  44. 44
    Xie, Y.; Chen, Y.; Ahmed, K. A.; Li, W.; Ahmed, S.; Sami, A.; Chibbar, R.; Tang, X.; Tao, M.; Xu, J.; Xiang, J. Potent CD4+ T-Cell Epitope P30 Enhances HER2/Neu-Engineered Dendritic Cell-Induced Immunity against Tg1-1 Breast Cancer in Transgenic FVBneuN Mice by Enhanced CD4+ T-Cell-Stimulated CTL Responses. Cancer Gene Ther. 2013, 20, 590– 598,  DOI: 10.1038/cgt.2013.60
    Google Scholar
    44
    Potent CD4+ T-cell epitope P30 enhances HER2/neu-engineered dendritic cell-induced immunity against Tg1-1 breast cancer in transgenic FVBneuN mice by enhanced CD4+ T-cell-stimulated CTL responses
    Xie, Y.; Chen, Y.; Ahmed, K. A.; Li, W.; Ahmed, S.; Sami, A.; Chibbar, R.; Tang, X.; Tao, M.; Xu, J.; Xiang, J.
    Cancer Gene Therapy (2013), 20 (10), 590-598CODEN: CGTHEG; ISSN:0929-1903. (Nature Publishing Group)
    One of the major obstacles in human epidermal growth factor receptor (HER)-2/neu-specific trastuzumab immunotherapy of HER2/neu-pos. breast cancer is the development of trastuzumab resistance, warranting the search for other therapeutic strategies. Although dendritic cell (DC) vaccines have been extensively applied in clin. trials for cancer treatment, the vaccination efficacy is still limited, mostly because DC vaccines are not sufficient to break tumor-assocd. antigen-specific self-immune tolerance in cancer patients. P30 (FNNFTVSFWLRVPKVSASHLE) derived from tetanus toxin is a universally potent CD4+ T helper epitope capable of enhancing CD8+ cytotoxic T-lymphocyte (CTL) responses. In this study, the authors constructed two recombinant adenoviral vectors (AdVs), AdVOVA-P30 and AdVHER2/neu-P30, expressing ovalbumin (OVA)-P30 and HER2/neu-P30. To enhance DC vaccine efficacy, the authors transfected mouse bone marrow (BM)-derived DCs with AdVOVA-P30 and AdVHER2/neu-P30 to generate engineered DCOVA-P30 and DCHER2/neu-P30 vaccines, resp. The authors, then, compared CD4+ and CD8+ T-cell responses and antitumor immunity derived from DCOVA-P30 and DCHER2/neu-P30 vaccination in wild-type C57BL/6 and transgenic FVBneuN mice, resp. The authors demonstrate that engineered DCOVA-P30 vaccine stimulates more efficient CD4+ and CD8+ T-cell responses than DCOVA in C57BL/6 mice. Interestingly, the increased DCOVA-P30-induced CTL responses are mainly contributed by enhanced CD4+ T-cell-stimulated CTL proliferation. The authors show that DCOVA-P30 vaccine also stimulates more efficient therapeutic immunity against OVA-expressing BL6-10OVA melanoma than DCOVA in C57BL/6 mice. In addn., the authors demonstrate that DCHER2/neu-P30 vaccine stimulates more efficient CD4+ and CD8+ T-cell responses and protective immunity against HER2/neu-expressing Tg1-1 breast cancer than DCHER2/neu in transgenic FVBneuN mice with HER2/neu-specific self-immune tolerance. Therefore, the engineered DCHER2/neu-P30 vaccine may provide a new immunotherapy alternative for women with HER2/neu+ breast cancer, esp. for trastuzumab-resistant HER2/neu+ breast cancer patients.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVynurvK&md5=41b0969396d251e48b29472f55e361df
  45. 45
    Pett, C.; Schorlemer, M.; Westerlind, U. A Unified Strategy for the Synthesis of Mucin Cores 1-4 Saccharides and the Assembled Multivalent Glycopeptides. Chem.─Eur. J. 2013, 19, 17001– 17010,  DOI: 10.1002/chem.201302921
    Google Scholar
    45
    A unified strategy for the synthesis of mucin cores 1-4 saccharides and the assembled multivalent glycopeptides
    Pett, Christian; Schorlemer, Manuel; Westerlind, Ulrika
    Chemistry - A European Journal (2013), 19 (50), 17001-17010CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
    By displaying different O-glycans in a multivalent mode, mucin and mucin-like glycoproteins are involved in a plethora of protein binding events. The understanding of the roles of the glycans and the identification of potential glycan binding proteins are major challenges. To enable future binding studies of mucin glycan and glycopeptide probes, a method that gives flexible and efficient access to all common mucin core-glycosylated amino acids was developed. Based on a convergent synthesis strategy starting from a shared early stage intermediate by differentiation in the glycoside acceptor reactivity, a common disaccharide building block allows for the creation of extended glycosylated amino acids carrying the mucin type-2 cores 1-4 saccharides. Formation of a phenyl-sulfenyl-N-Troc (Troc = trichloroethoxycarbonyl) byproduct during N-iodosuccinimide-promoted thioglycoside couplings was further characterized and a new methodol. for the removal of the Troc group is described. The obtained glycosylated 9-fluorenylmethoxycarbonyl (Fmoc)-protected amino acid building blocks are incorporated into peptides for multivalent glycan display.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhslaqsL3E&md5=76a4e68e8e97ba1fc418a5a6e3968d89
  46. 46
    Pett, C.; Westerlind, U. A Convergent Strategy for the Synthesis of Type-1 Elongated Mucin Cores 1-3 and the Corresponding Glycopeptides. Chem.─Eur. J. 2014, 20, 7287– 7299,  DOI: 10.1002/chem.201400162
    Google Scholar
    46
    A Convergent Strategy for the Synthesis of Type-1 Elongated Mucin Cores 1-3 and the Corresponding Glycopeptides
    Pett, Christian; Westerlind, Ulrika
    Chemistry - A European Journal (2014), 20 (24), 7287-7299CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Mucins are a class of highly O-glycosylated proteins found on the surface of cells in epithelial tissues. O-Glycosylation is crucial for the functionality of mucins and changes therein can have severe consequences for an organism. With that in mind, the elucidation of interactions of carbohydrate binding proteins with mucins, whether in morbidly altered or unaltered conditions, continue to shed light on mechanisms involved in diseases like chronic inflammations and cancer. Despite the known importance of type-1 and type-2 elongated mucin cores 1-4 in glycobiol., the corresponding type-1 structures are much less well studied. Here, the first chem. synthesis of extended mucin type-1 O-glycan core 1-3 amino acid structures based on a convergent approach is presented. By utilizing differentiation in acceptor reactivity, shared early stage Tn- and T-acceptor intermediates were elongated with a common type-1 [β-D-Gal-1,3-β-D-GlcNAc] disaccharide, which allows for straightforward prepn. of diverse glycosylated amino acids carrying the type-1 mucin core 1-3 saccharides. The obtained glycosylated 9-fluorenylmethoxycarbonyl (Fmoc)-protected amino acid building blocks were employed in synthesis of type-1 mucin glycopeptides, which are useful in biol. applications.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXot1Sitbc%253D&md5=7ae3edc6aada767e8621be31268fd418
  47. 47
    Li, R.-J. E.; Hogervorst, T. P.; Achilli, S.; Bruijns, S. C. M.; Spiekstra, S.; Vivès, C.; Thépaut, M.; Filippov, D. V.; van der Marel, G. A.; van Vliet, S. J.; Fieschi, F.; Codée, J. D. C.; van Kooyk, Y. Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens. Front. Cell Dev. Biol. 2020, 8, 556,  DOI: 10.3389/fcell.2020.00556
    Google Scholar
    47
    Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens
    Li Rui-Jun Eveline; Bruijns Sven C M; Spiekstra Sander; van Vliet Sandra J; van Kooyk Yvette; Hogervorst Tim P; Filippov Dmitri V; van der Marel Gijs A; Codee Jeroen D C; Achilli Silvia; Vives Corinne; Thepaut Michel; Fieschi Franck
    Frontiers in cell and developmental biology (2020), 8 (), 556 ISSN:2296-634X.
    Langerhans cells (LCs) are antigen-presenting cells that reside in the skin. They uniquely express high levels of the C-type lectin receptor Langerin (CD207), which is an attractive target for antigen delivery in immunotherapeutic vaccination strategies against cancer. We here assess a library of 20 synthetic, well-defined mannoside clusters, built up from one, two, and three of six monomannosides, dimannosides, or trimannosides, appended to an oligopeptide backbone, for binding with Langerin using surface plasmon resonance and flow cytometric quantification. It is found that Langerin binding affinity increases with increasing number of mannosides. Hexavalent presentation of the mannosides resulted in binding affinities ranging from 3 to 12 μM. Trivalent presentation of the dimannosides and trimannosides led to Langerin affinity in the same range. The model melanoma gp100 antigenic peptide was subsequently equipped with a hexavalent cluster of the dimannosides and trimannosides as targeting moieties. Surprisingly, although the bifunctional conjugates were taken up in LCs in a Langerin-dependent manner, limited antigen presentation to cytotoxic T cells was observed. These results indicate that targeting glycan moieties on immunotherapeutic vaccines should not only be validated for target binding, but also on the continued effects on biology, such as antigen presentation to both CD8(+) and CD4(+) T cells.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB38fitVaqtA%253D%253D&md5=d0aefd6bb1ad3f4b602a33cb65d76342
  48. 48
    Li, R.-J. E.; Hogervorst, T. P.; Achilli, S.; Bruijns, S. C.; Arnoldus, T.; Vivès, C.; Wong, C. C.; Thépaut, M.; Meeuwenoord, N. J.; van den Elst, H.; Overkleeft, H. S.; van der Marel, G. A.; Filippov, D. V.; van Vliet, S. J.; Fieschi, F.; Codée, J. D. C.; van Kooyk, Y. Systematic Dual Targeting of Dendritic Cell C-Type Lectin Receptor DC-SIGN and TLR7 Using a Trifunctional Mannosylated Antigen. Front. Chem. 2019, 7, 650,  DOI: 10.3389/fchem.2019.00650
    Google Scholar
    48
    Systematic dual targeting of dendritic cell C-type lectin receptor DC-SIGN and TLR7 using a trifunctional mannosylated antigen
    Li, Rui-Jun Eveline; Hogervorst, Tim P.; Achilli, Silvia; Bruijns, Sven C.; Arnoldus, Tim; Vives, Corinne; Wong, Chung C.; Thepaut, Michel; Meeuwenoord, Nico J.; van den Elst, Hans; Overkleeft, Herman S.; van der Marel, Gijs A.; Filippov, Dmitri V.; van Vliet, Sandra J.; Fieschi, Franck; Codee, Jeroen D. C.; van Kooyk, Yvette
    Frontiers in Chemistry (Lausanne, Switzerland) (2019), 7 (), 650CODEN: FCLSAA; ISSN:2296-2646. (Frontiers Media S.A.)
    Dendritic cells are important initiators of adaptive immunity, and they possess a multitude of Pattern Recognition Receptors to generate an adequate T cell mediated immunity against invading pathogens. PRR ligands are frequently conjugated to tumor-assocd. antigens in a vaccination strategy to enhance the immune response toward such antigens. One of these PPRs, DC-SIGN, a member of the C-type lectin receptor family, has been extensively targeted with Lewis structures and mannose glycans, often presented in multivalent fashion. Hexavalent presentation of the clusters showed the highest binding affinity, with the hexa-α1,2-di-mannoside being the most potent ligand. The four highest binding hexavalent mannoside structures were conjugated to a model melanoma gp100-peptide antigen and further equipped with a Toll-like receptor 7 (TLR7)-agonist as adjuvant for DC maturation, creating a trifunctional vaccine conjugate. Interestingly, DC-SIGN affinity of the mannoside clusters did not directly correlate with antigen presentation enhancing properties and the α1,2-di-mannoside cluster with the highest binding affinity in our library even hampered T cell activation. Overall, this systematic study has demonstrated that multivalent glycan presentation can improve DC-SIGN binding but enhanced binding cannot be directly translated into enhanced antigen presentation and the sole assessment of binding affinity is thus insufficient to det. further functional biol. activity.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXps1Crs7w%253D&md5=1857ff8a25c82901090efae7c11e215a
  49. 49
    Wu, X.; McKay, C.; Pett, C.; Yu, J.; Schorlemer, M.; Ramadan, S.; Lang, S.; Behren, S.; Westerlind, U.; Finn, M. G.; Huang, X. Synthesis and Immunological Evaluation of Disaccharide Bearing MUC-1 Glycopeptide Conjugates with Virus-like Particles. ACS Chem. Biol. 2019, 14, 2176– 2184,  DOI: 10.1021/acschembio.9b00381
    Google Scholar
    49
    Synthesis and immunological evaluation of disaccharide bearing MUC-1 glycopeptide conjugates with virus-like particles
    Wu, Xuanjun; McKay, Craig; Pett, Christian; Yu, Jin; Schorlemer, Manuel; Ramadan, Sherif; Lang, Shuyao; Behren, Sandra; Westerlind, Ulrika; Finn, M. G.; Huang, Xuefei
    ACS Chemical Biology (2019), 14 (10), 2176-2184CODEN: ACBCCT; ISSN:1554-8929. (American Chemical Society)
    Mucin-1 (MUC1) is a highly attractive antigenic target for anticancer vaccines. Naturally existing MUC1 can contain multiple types of O-linked glycans, including the Thomsen-Friedenreich (Tf) antigen and the Sialyl Thomsen-nouveau (STn) antigen. In order to target these antigens as potential anticancer vaccines, MUC1 glycopeptides SAPDT*RPAP (T* is the glycosylation site) bearing the Tf and the STn antigen, resp., have been synthesized. The bacteriophage Qβ carrier is a powerful carrier for antigen delivery. The conjugates of MUC1-Tf and -STn glycopeptides with Qβ were utilized to immunize immune-tolerant human MUC1 transgenic (MUC1.Tg) mice, which elicited superior levels of anti-MUC1 IgG antibodies with titers reaching over 2 million units. The IgG antibodies recognized a wide range of MUC1 glycopeptides bearing diverse glycans. Antibodies induced by Qβ-MUC1-Tf showed strongest binding, with MUC1-expressing melanoma B16-MUC1 cells, and effectively killed these cells in vitro. Vaccination with Qβ-MUC1-Tf first followed by tumor challenge in a lung metastasis model showed significant redns. of the no. of tumor foci in the lungs of immunized mice as compared to those in control mice. This was the first time that a MUC1-Tf-based vaccine has shown in vivo efficacy in a tumor model. As such, Qβ-MUC1 glycopeptide conjugates have great potential as anticancer vaccines.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhslegtL3E&md5=bdf9d26dd58bd13745dc5219ff444017
  50. 50
    Pirro, M.; Schoof, E.; Van Vliet, S. J.; Rombouts, Y.; Stella, A.; De Ru, A.; Mohammed, Y.; Wuhrer, M.; Van Veelen, P. A.; Hensbergen, P. J. Glycoproteomic Analysis of MGL-Binding Proteins on Acute T-Cell Leukemia Cells. J. Proteome Res. 2019, 18, 1125– 1132,  DOI: 10.1021/acs.jproteome.8b00796
    Google Scholar
    50
    Glycoproteomic Analysis of MGL-Binding Proteins on Acute T-Cell Leukemia Cells
    Pirro, Martina; Schoof, Esmee; van Vliet, Sandra J.; Rombouts, Yoann; Stella, Alexandre; de Ru, Arnoud; Mohammed, Yassene; Wuhrer, Manfred; van Veelen, Peter A.; Hensbergen, Paul J.
    Journal of Proteome Research (2019), 18 (3), 1125-1132CODEN: JPROBS; ISSN:1535-3893. (American Chemical Society)
    C-type lectins are a diverse group of proteins involved in many human physiol. and pathol. processes. Most C-type lectins are glycan-binding proteins, some of which are pivotal for innate immune responses against pathogens. Other C-type lectins, such as the macrophage galactose-type lectin (MGL), have been shown to induce immunosuppressive responses upon the recognition of aberrant glycosylation on cancer cells. MGL is known to recognize terminal N-acetylgalactosamine (GalNAc), such as the Tn antigen, which is commonly found on malignant cells. Even though this glycan specificity of MGL is well described, there is a lack of understanding of the actual glycoproteins that bind MGL. We present a glycoproteomic workflow for the identification of MGL-binding proteins, which we applied to study MGL ligands on the human Jurkat leukemia cell line. In addn. to the known MGL ligands and Tn antigen-carrying proteins CD43 and CD45 on these cells, we have identified a set of novel cell-surface ligands for MGL. Importantly, for several of these, O-glycosylation has hitherto not been described. Altogether, our data provide new insight into the identification and structure of novel MGL ligands that presumably act as modulatory mols. in cancer immune responses.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXisFyksrjE&md5=b9d3814d8c244b0c9528e4dc6b3370f6
  51. 51
    Seder, R.; Reed, S. G.; O’Hagan, D.; Malyala, P.; D’Oro, U.; Laera, D.; Abrignani, S.; Cerundolo, V.; Steinman, L.; Bertholet, S. Gaps in Knowledge and Prospects for Research of Adjuvanted Vaccines. Vaccine 2015, 33, B40– B43,  DOI: 10.1016/j.vaccine.2015.03.057
    Google Scholar
    There is no corresponding record for this reference.
  52. 52
    Hanisch, F.-G.; Stadie, T.; Deutzmann, F.; Peter-Katalinic, J. MUC1 Glycoforms in Breast Cancer. Cell Line T47D as a Model for Carcinoma-Associated Alterations of O-Glycosylation. Eur. J. Biochem. 1996, 236, 318– 327,  DOI: 10.1111/j.1432-1033.1996.00318.x
    Google Scholar
    52
    MUC1 glycoforms in breast cancer. Cell line T47D as a model for carcinoma-associated alterations of O-glycosylation
    Hanisch, Franz-Georg; Stadie, Tanja R. E.; Deutzmann, Frank; Peter-Katalinic, Jasna
    European Journal of Biochemistry (1996), 236 (1), 318-27CODEN: EJBCAI; ISSN:0014-2956. (Springer)
    A highly immunogenic peptide motif within the tandem repeat domain of MUC1 mucin is assumed to be exposed during development of breast cancer due to altered O-glycosylation. To elucidate the structural aspects of these changes, the authors have isolated and analyzed the integrated or secretory MUC1 glycoforms from carcinoma cell lines or solid tumors and from human milk. The buoyant densities measured in CsCl gradients for MUC1 glycoforms from cancer cells revealed heterogeneity of the physicochem. species and a significant redn. of their carbohydrate contents compared to MUC1 from skim milk. Immunoreactivity patterns of MUC1 glycoforms from tumor or T47D cells exhibited a lack of fucosylated Lewis blood-group-related antigens and the appearance of core-type antigen sialyl-(NeuGl)-TF, Galβ1-3(NeuGlα2-6)GalNAc. Structural chem. of MUC1 oligosaccharides demonstrated that the cancer-assocd. glycoforms carry mainly sialylated trisaccharides NeuAcα2-3Galβ1-3GalNAc or NeuAcα2-6(Galβ1-3)GalNAc, exhibit a concomitant decrease in the ratio of GlcNAc/GalNAc, a redn. or disappearance of L-fucose, and a partial substitution of N-acetylneuraminic acid by the N-glycolylated variant. On comparison to the secretory MUC1 in human milk, the glycoforms on human milk fat globule membranes showed apparently identical patterns of O-linked oligosaccharides with a preponderance of neutral polylactosamino-glycans. During serum-free cultivation of T47D cells over 4 wk, the expression of secretory MUC1 glycoforms was inconsistent based on the decreasing contents of sialic acid and on the concomitant increase of immunodetectable TF antigen.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28Xhtlantrs%253D&md5=24687001dc9ac24202033c516bf4ec5a
  53. 53
    Stergiou, N.; Nagel, J.; Pektor, S.; Heimes, A. S.; Jäkel, J.; Brenner, W.; Schmidt, M.; Miederer, M.; Kunz, H.; Roesch, F.; Schmitt, E. Evaluation of a Novel Monoclonal Antibody against Tumor-Associated MUC1 for Diagnosis and Prognosis of Breast Cancer. Int. J. Med. Sci. 2019, 16, 1188– 1198,  DOI: 10.7150/ijms.35452
    Google Scholar
    53
    Evaluation of a novel monoclonal antibody against tumor-associated MUC1 for diagnosis and prognosis of breast cancer
    Stergiou, Natascha; Nagel, Johannes; Pektor, Stefanie; Heimes, Anne-Sophie; Jaekel, Joerg; Brenner, Walburgis; Schmt, Marcus; Miederer, Matthias; Kunz, Horst; Roesch, Frank; Schmitt, Edgar
    International Journal of Medical Sciences (2019), 16 (9), 1188-1198CODEN: IJMSGZ; ISSN:1449-1907. (Ivyspring International Publisher)
    There is still a great unmet medical need concerning diagnosis and treatment of breast cancer which could be addressed by utilizing specific mol. targets. Tumor-assocd. MUC1 is expressed on over 90 % of all breast cancer entities and differs strongly from its physiol. form on epithelial cells, therefore presenting a unique target for breast cancer diagnosis and antibody-mediated immune therapy. Utilizing an anti-tumor vaccine based on a synthetically prepd. glycopeptide, we generated a monoclonal antibody (mAb) GGSK-1/30, selectively recognizing human tumor-assocd. MUC1. This antibody targets exclusively tumor-assocd. MUC1 in the absence of any binding to MUC1 on healthy epithelial cells thus enabling the generation of breast tumor-specific radiolabeled immune therapeutic tools. Methods: MAb GGSK-1/30 was used for immunohistochem. anal. of human breast cancer tissue. Its desferrioxamine (Df')-conjugate was synthesized and labeled with 89Zr. [89Zr]Zr-Df'-GGSK-1/30 was evaluated as a potential PET tracer. Binding and pharmacokinetic properties of [89Zr]Zr-Df'-GGSK-1/30 were analyzed in vitro using human and murine cell lines that express tumor-assocd. MUC1. Self-generated primary murine breast cancer cells expressing human tumor-assocd. MUC1 were transplanted s.c. in wild type and human MUC1-transgenic mice. The pharmacol. of [89Zr]Zr-Df'-GGSK-1/30 was investigated using breast tumor-bearing mice in vivo by PET/MRT imaging as well as by ex vivo organ biodistribution anal. Results: The mAb GGSK-1/30 stained specifically human breast tumor tissue and can be possibly used to predict the severity of disease progression based on the expression of the tumor-assocd. MUC1. For in vivo imaging, the Df'-conjugated mAb was radiolabeled with a radiochem. yield of 60 %, a radiochem. purity of 95 % and an apparent specific activity of 6.1 GBq/μmol. After 7 d, stabilities of 84 % in human serum and of 93 % in saline were obsd. In vitro cell studies showed strong binding to human tumor-assocd. MUC1 expressing breast cancer cells. The breast tumor-bearing mice showed an in vivo tumor uptake of >50 %ID/g and clearly visible specific enrichment of the radioconjugate via PET/MRT. Principal conclusions: Tumor-assocd. MUC1 is a very important biomarker for breast cancer next to the traditional markers estrogen receptor (ER), progesterone receptor (PR) and HER/2-neu. The mAb GGSK-1/30 can be used for the diagnosis of over 90% of breast cancers, including triple neg. breast cancer based on biopsy staining. Its radioimmunoconjugate represents a promising PET-tracer for breast cancer imaging selectively targeting breast cancer cells.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXlsVKmsbk%253D&md5=dbca95f8f30a8df92aec9a049a488e4c
  54. 54
    Festari, M. F.; Da Costa, V.; Rodríguez-Zraquia, S. A.; Costa, M.; Landeira, M.; Lores, P.; Solari-Saquieres, P.; Kramer, M. G.; Freire, T. The Tumor-Associated Tn Antigen Fosters Lung Metastasis and Recruitment of Regulatory T Cells in Triple Negative Breast Cancer. Glycobiology 2022, 32, 366– 379,  DOI: 10.1093/glycob/cwab123
    Google Scholar
    54
    The tumor-associated Tn antigen fosters lung metastasis and recruitment of regulatory T cells in triple negative breast cancer
    Festari, Maria Florencia; da Costa, Valeria; Rodriguez-Zraquia, Santiago A.; Costa, Monique; Landeira, Mercedes; Lores, Pablo; Solari-Saquieres, Patricia; Kramer, M. Gabriela; Freire, Teresa
    Glycobiology (2022), 32 (5), 366-379CODEN: GLYCE3; ISSN:1460-2423. (Oxford University Press)
    Cancer is a leading cause of death worldwide, accounting for nearly 10 million deaths. Among breast cancers (BC) subtypes, triple-neg. (TN) BC is characterized by metastatic progression and poor patient prognosis. Although, TNBC is initially sensitive to chemotherapy, many TNBC patients rapidly develop resistance, at which point metastatic disease is highly lethal. Cancer cells present phenotypic changes or mol. signatures that distinguish them from healthy cells. The Tn antigen (GalNAc-O-Thr/Ser), which constitutes a powerful tool as tumor marker, was recently reported to contribute to tumor growth. However, its role in BC-derived metastasis has not yet been addressed. In this work, we generated a pre-clin. orthotopic Tn+ model of metastatic TNBC, which mimics the patient surgical treatment and is useful to study the role of Tn in metastasis and immunoregulation. We obtained two different cell clones, which differed in their Tn antigen expression: a high Tn-expressing and a non-expressing clone. Interestingly, the Tn-pos. cell line generated significantly larger tumors and higher degree of lung metastases assocd. with a lower survival rate than the Tn-neg. and parental cell line. Furthermore, we also found that both tumors and draining-lymph nodes from Tn+-tumor-bearing mice presented a higher frequency of CD4+ FoxP3+ T cells, while their splenocytes expressed higher levels of IL-10. In conclusion, this work suggests that the Tn antigen participates in breast tumor growth and spreading, favoring metastases to the lungs that are assocd. with an immunoregulatory state, suggesting that Tn-based immunotherapy could be a strategy of choice to treat these tumors.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XisFKmtb3I&md5=66668ef3e2760c1ecacdea50dfbd8fc4
  55. 55
    Khosrowabadi, E.; Wenta, T.; Keskitalo, S.; Manninen, A.; Kellokumpu, S. Altered Glycosylation of Several Metastasis-Associated Glycoproteins with Terminal GalNAc Defines the Highly Invasive Cancer Cell Phenotype. Oncotarget 2022, 13, 73– 89,  DOI: 10.18632/oncotarget.28167
    Google Scholar
    55
    Altered glycosylation of several metastasis-associated glycoproteins with terminal GalNAc defines the highly invasive cancer cell phenotype
    Khosrowabadi Elham; Wenta Tomasz; Manninen Aki; Kellokumpu Sakari; Keskitalo Salla
    Oncotarget (2022), 13 (), 73-89 ISSN:.
    Several distinct metastasis-associated glycosylation changes have been shown to promote cancer cell invasion and metastasis, the main cause of death of cancer patients. However, it is unclear whether their presence reflects cell- or tissue-specific variations for metastasis, or species needed to drive different phases of the metastatic cascade. To address this issue from a different perspective, we investigated here whether different cancer cell lines share any glycotopes that are common and important for their invasive phenotype. By using lectin microarray glycan profiling and an established myoma tissue-based 3D invasion assay, we identified a single glycotope recognized by Helix Pomatia agglutinin (HPA), whose expression level in different cancer cells correlated significantly with their invasive potential. Lectin pull-down assay and LC-MS/MS analysis in highly- (A431 and SW-48) and poorly invasive (HepG2 and RCC4) cancer cells revealed ~85 glycoproteins of which several metastasis-promoting members of the integrin family of cell adhesion receptors, the epidermal growth factor receptor (EGFR) and the matrix metalloproteinase-14 (MMP-14) were among the abundant ones. Moreover, we showed that the level of the GalNAc glycotope in MMP-14, EGFR, αV-, β1- and β4 integrin in highly and poorly invasive cancer cells correlated positively with their invasive potential. Collectively, our findings suggest that altered glycosylation of several metastasis-associated glycoproteins with terminal GalNAc drives the highly invasive cancer cell phenotype.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB2M%252FotVagtQ%253D%253D&md5=f64cd9793dd2699928fc68aa5b362687
  56. 56
    Geijtenbeek, T. B. H.; Gringhuis, S. I. Signalling through C-Type Lectin Receptors: Shaping Immune Responses. Nat. Rev. Immunol. 2009, 9, 465– 479,  DOI: 10.1038/nri2569
    Google Scholar
    56
    Signalling through C-type lectin receptors: shaping immune responses
    Geijtenbeek, Teunis B. H.; Gringhuis, Sonja I.
    Nature Reviews Immunology (2009), 9 (7), 465-479CODEN: NRIABX; ISSN:1474-1733. (Nature Publishing Group)
    A review. C-type lectin receptors (CLRs) expressed by dendritic cells are crucial for tailoring immune responses to pathogens. Following pathogen binding, CLRs trigger distinct signalling pathways that induce the expression of specific cytokines which det. T cell polarization fates. Some CLRs can induce signalling pathways that directly activate nuclear factor-κB, whereas other CLRs affect signalling by Toll-like receptors. Dissecting these signalling pathways and their effects on host immune cells is essential to understand the mol. mechanisms involved in the induction of adaptive immune responses. In this Review we describe the role of CLR signalling in regulating adaptive immunity and immunopathogenesis and discuss how this knowledge can be harnessed for the development of innovative vaccination approaches.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXntFCgurc%253D&md5=91f3810595e1bb4794efa63a97321ecf

Cited By

Click to copy section linkSection link copied!

This article is cited by 1 publications.

  1. Shi-Hao Zhou, Ru-Yan Zhang, Yu Wen, Yong-Ke Zou, Dong Ding, Miao-Miao Bian, Hong-Ying Cui, Jun Guo. Multifunctional Lipidated Protein Carrier with a Built-In Adjuvant as a Universal Vaccine Platform Potently Elevates Immunogenicity of Weak Antigens. Journal of Medicinal Chemistry 2024, 67 (8) , 6822-6838. https://doi.org/10.1021/acs.jmedchem.4c00412
Download PDF
Go to Journal of the American Chemical Society
Get e-Alerts
Get e-Alerts

Journal of the American Chemical Society

Cite this: J. Am. Chem. Soc. 2023, 145, 24, 13027–13037
Click to copy citationCitation copied!
https://doi.org/10.1021/jacs.2c12843
Published June 6, 2023

Copyright © 2023 The Authors. Published by American Chemical Society. This publication is licensed under

CC-BY 4.0 .

Article Views

8283

Altmetric

-

Citations

1
Learn about these metrics

Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.

  • Abstract

    High Resolution Image
    Download MS PowerPoint Slide

    Scheme 1

    Scheme 1. Synthesis of the Trivalent Ligand 1 to Engage MGL
    High Resolution Image
    Download MS PowerPoint Slide

    Figure 1

    Figure 1. Summary of peptides 9–16 synthesized by SPPS.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 2

    Figure 2. EC50 values for the Tn-MUC1 (glyco)peptides and their glycocluster-conjugates with MGL were obtained using ELISA assays of a dilution series of the immobilized peptides 9–14 and recombinant MGL: the biotinylated peptides 9–14 were coupled to a streptavidin-coated plate at twofold dilutions, which was then titrated with recombinant MGL-Fc. MGL binding was detected using peroxidase conjugated to a goat anti-human Fc Ab. Curve fitting is shown in the figure. For all the peptides, the R2 value of the curve fitting was above 0.98.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 3

    Figure 3. Flow cytometry analysis on BMDCs. (A) MFI of MHCII+CD11c+ cells relative to the peptides. BMDCs where incubated in the presence of the biotinylated peptides 10, 11, and 12 at 4 °C and then labeled with streptavidine-PE. (B) Peptides-associated fluorescence intensity histogram after incubation with BMDCs at 37 °C. (C) MFI relative to the MGL receptor on BMDC surface. BMDCs were treated with peptide 12 for 30 min at 37 °C. Error bars represent standard errors.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 4

    Figure 4. Comparison of fluorophore-labeled MUC1 peptides 10 and 12 by microscopy. BMDCs were treated with 5 μg/mL of the biotinylated peptides 10 and 12 at 4 °C for 1 h followed by washing and treatment with streptavidin-Alexa 594. (A,B) Localization of the peptides on the cells surface. (C) Localization of the peptides after incubation for 30 min and 1 h at 37 °C. Internalization was visualized by confocal microscopy.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 5

    Figure 5. In vivo experiment immunization schedule and antibody analysis. (A) Experimental design: 2.5 nmol of construct per injection; day 0, immunization subcutaneous (s.c.) with complete Freund’s adjuvant (CFA); day s14, 28, and 42, immunization intraperitoneal (i.p.) with incomplete Freund’s Adjuvant (IFA); blood collection: 5 days post immunization. (B) IgG titers in the serum of mice immunized with constructs 15 and 16, respectively. The sera were collected at days 19, 33, and 47. (C) Total IgG and IgG subtypes from sera collected at day 47 from mice immunized with constructs 15 and 16. All ELISA measurements were performed on plates coated with BSA-(11)4-17 synthesized as reported in the Supporting Information. The p values are determined through two-way ANOVA with Šídák multiple comparison test using GraphPad Prism, ***p < 0.001; error bars indicate standard error.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 6

    Figure 6. Flow cytometry analysis of the specific recognition of breast tumor cells T47D by antibodies. (A) MFIs of binding of T47D-MUC1 cells by sera (1:100) of three mice (red, green, and blue curves) immunized with construct 16. The gray filled trace is from pre-immune serum. (B) Comparison of the MFIs of antibodies from sera of 15 and 16 (1:100). Error bars represent ±1 standard error from the mean of three biological replicates (n = 3). MUC1 expression on TD47 was confirmed by anti-MUC1 mAb HPMV (1:5 dilution) produced in-house. (53) The p-values were determined through a two-tailed t-test using GraphPad Prism, **p < 0.01; error bars indicate standard error.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 7

    Figure 7. Binding specificity of antibodies generated by vaccines 15 and 16 to mucin core Tn antigen, T antigen, and 1 and 3 glycopeptides. Binding of sera at 1:1000 dilution to an array of varied mucin peptide sequences, glycosylation sites, and glycan structures are shown. Mouse sera derived from 15 and 16 showed similar recognition patterns. However, higher binding to MUC1 glycopeptides was observed for mice immunized with 16. For complete figures, see S14 and S15. Bars represent standard error (n = 5).

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 8

    Figure 8. Binding specificity of antibodies generated by vaccines 15 and 16 to mucin core 2 and 4 glycopeptides. Binding of sera at 1:1000 dilution to an array of varied mucin peptide sequences, glycosylation sites, and glycan structures are shown. Mouse sera derived from 15 and 16 showed preferential binding to tumor-associated mucin core. For complete figures, see S14 and S15. Bars represent standard error (n = 5).

    High Resolution Image
    Download MS PowerPoint Slide

  • References

    This article references 56 other publications.

    1. 1
      van Vliet, S. J.; Saeland, E.; van Kooyk, Y. Sweet Preferences of MGL: Carbohydrate Specificity and Function. Trends Immunol. 2008, 29, 83– 90,  DOI: 10.1016/j.it.2007.10.010
      1
      Sweet preferences of MGL: carbohydrate specificity and function
      van Vliet, Sandra J.; Saeland, Eirikur; van Kooyk, Yvette
      Trends in Immunology (2008), 29 (2), 83-90CODEN: TIRMAE; ISSN:1471-4906. (Elsevier B.V.)
      A review. C-type lectins play important roles in both innate and adaptive immune responses. In contrast to the mannose- or fucose-specific C-type lectins DC-SIGN and mannose receptor, the galactose-type lectins, of which only macrophage galactose-type lectin (MGL) is found within the immune system, are less well known. MGL is selectively expressed by immature dendritic cells and macrophages with elevated levels on tolerogenic or alternatively activated subsets. Human MGL has an exclusive specificity for rare terminal GalNAc structures, which are revealed on the tumor-assocd. mucin MUC1 and CD45 on effector T cells. These findings implicate MGL in the homeostatic control of adaptive immunity. The authors discuss here the functional similarities and differences between MGL orthologs and compare MGL to its closest homolog, the liver-specific asialoglycoprotein receptor (ASGP-R).
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXhslGitLc%253D&md5=a79c991afa6912a3da6a2f349682dcb6
    2. 2
      van Vliet, S. J.; van Liempt, E.; Saeland, E.; Aarnoudse, C. A.; Appelmelk, B.; Irimura, T.; Geijtenbeek, T. B. H.; Blixt, O.; Alvarez, R.; van Die, I.; van Kooyk, Y. Carbohydrate Profiling Reveals a Distinctive Role for the C-Type Lectin MGL in the Recognition of Helminth Parasites and Tumor Antigens by Dendritic Cells. Int. Immunol. 2005, 17, 661– 669,  DOI: 10.1093/intimm/dxh246
      2
      Carbohydrate profiling reveals a distinctive role for the C-type lectin MGL in the recognition of helminth parasites and tumor antigens by dendritic cells
      van Vliet, Sandra J.; van Liempt, Ellis; Saeland, Eirikur; Aarnoudse, Corlien A.; Appelmelk, Ben; Irimura, Tatsuro; Geijtenbeek, Teunis B. H.; Blixt, Ola; Alvarez, Richard; van Die, Irma; van Kooyk, Yvette
      International Immunology (2005), 17 (5), 661-669CODEN: INIMEN; ISSN:0953-8178. (Oxford University Press)
      Dendritic cells (DCs) are key to the maintenance of peripheral tolerance to self-antigens and the orchestration of an immune reaction to foreign antigens. C-type lectins, expressed by DCs, recognize carbohydrate moieties on antigens that can be internalized for processing and presentation. Little is known about the exact glycan structures on self-antigens and pathogens that are specifically recognized by the different C-type lectins and how this interaction influences DC function. The authors have analyzed the carbohydrate specificity of the human C-type lectin macrophage galactose-type lectin (MGL) using glycan microarray profiling and identified an exclusive specificity for terminal α- and β-linked GalNAc residues that naturally occur as parts of glycoproteins or glycosphingolipids. Specific glycan structures contg. terminal GalNAc moieties, expressed by the human helminth parasite Schistosoma mansoni as well as tumor antigens and a subset of gangliosides, were identified as ligands for MGL. The authors' results indicate an endogenous function for DC-expressed MGL in the clearance and tolerance to self-gangliosides, and in the pattern recognition of tumor antigens and foreign glycoproteins derived from helminth parasites.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXjslOqu70%253D&md5=8f8fc96e5d622e47338311d219077a3e
    3. 3
      Pirro, M.; Rombouts, Y.; Stella, A.; Neyrolles, O.; Burlet-Schiltz, O.; van Vliet, S. J.; de Ru, A. H.; Mohammed, Y.; Wuhrer, M.; van Veelen, P. A.; Hensbergen, P. J. Characterization of Macrophage Galactose-Type Lectin (MGL) Ligands in Colorectal Cancer Cell Lines. Biochim. Biophys. Acta, Gen. Subj. 2020, 1864, 129513,  DOI: 10.1016/j.bbagen.2020.129513
      3
      Characterization of Macrophage Galactose-type Lectin (MGL) ligands in colorectal cancer cell lines
      Pirro, Martina; Rombouts, Yoann; Stella, Alexandre; Neyrolles, Olivier; Burlet-Schiltz, Odile; van Vliet, Sandra J.; de Ru, Arnoud H.; Mohammed, Yassene; Wuhrer, Manfred; van Veelen, Peter A.; Hensbergen, Paul J.
      Biochimica et Biophysica Acta, General Subjects (2020), 1864 (4), 129513CODEN: BBGSB3; ISSN:0304-4165. (Elsevier B.V.)
      The Ca2+-dependent C-type lectin receptor Macrophage Galactose-type Lectin (MGL) is highly expressed by tolerogenic dendritic cells (DC) and macrophages. MGL exhibits a high binding specificity for terminal alpha- and beta-linked GalNAc residues found in Tn, sTn and LacdiNAc antigens. These glycan epitopes are often overexpressed in colorectal cancer (CRC), and, as such, MGL can be used to discriminate tumor from the corresponding healthy tissues. Moreover, the high expression of MGL ligands is assocd. with poor disease-free survival in stage III of CRC tumors. Nonetheless, the glycoproteins expressed by tumor cells that are recognized by MGL have hitherto remained elusive. Using a panel of three CRC cell lines (HCT116, HT29 and LS174T), recapitulating CRC diversity, we performed FACS staining and pull-down assays using a recombinant sol. form of MGL (and a mutant MGL as control) combined with mass spectrometry-based (glyco)proteomics. HCT116 and HT29, but not LS174T, are high MGL-binding CRC cell lines. On these cells, the major cell surface binding proteins are receptors (e.g. MET, PTK7, SORL1, PTPRF) and integrins (ITGB1, ITGA3). From these proteins, several N- and/or O-glycopeptides were identified, of which some carried either a LacdiNAc or Tn epitope. We have identified cell surface MGL-ligands on CRC cell lines. Advances in (glyco)proteomics have led to identification of candidate key mediators of immune-evasion and tumor growth in CRC.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhtFCktbw%253D&md5=d8254b636571303acda5e1c3491ea474
    4. 4
      Marcelo, F.; Supekar, N.; Corzana, F.; Van Der Horst, J. C.; Vuist, I. M.; Live, D.; Boons, G. J. P. H.; Smith, D. F.; Van Vliet, S. J. Identification of a Secondary Binding Site in Human Macrophage Galactose-Type Lectin by Microarray Studies: Implications for the Molecular Recognition of Its Ligands. J. Biol. Chem. 2019, 294, 1300– 1311,  DOI: 10.1074/jbc.ra118.004957
      4
      Identification of a secondary binding site in human macrophage galactose-type lectin by microarray studies: Implications for the molecular recognition of its ligands
      Marcelo, Filipa; Supekar, Nitin; Corzana, Francisco; van der Horst, Joost C.; Vuist, Ilona M.; Live, David; Boons, Geert-Jan P. H.; Smith, David F.; van Vliet, Sandra J.
      Journal of Biological Chemistry (2019), 294 (4), 1300-1311CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
      The human macrophage galactose-type lectin (MGL) is a C-type lectin characterized by a unique specificity for terminal GalNAc residues present in the tumor-assocd. Tn antigen (αGalNAc-Ser/Thr) and its sialylated form, the sialyl-Tn antigen. However, human MGL has multiple splice variants, and whether these variants have distinct ligand-binding properties is unknown. Here, using glycan microarrays, we compared the binding properties of the short MGL 6C (MGLshort) and the long MGL 6B (MGLlong) splice variants, as well as of a histidine-to-threonine mutant (MGLshort H259T). Although the MGLshort and MGLlong variants displayed similar binding properties on the glycan array, the MGLshort H259T mutant failed to interact with the sialyl-Tn epitope. As the MGLshort H259T variant could still bind a single GalNAc monosaccharide on this array, we next investigated its binding characteristics to Tn-contg. glycopeptides derived from the MGL ligands mucin 1 (MUC1), MUC2, and CD45. Strikingly, in the glycopeptide microarray, the MGLshort H259T variant lost high-affinity binding toward Tn-contg. glycopeptides, esp. at low probing concns. Moreover, MGLshort H259T was unable to recognize cancer-assocd. Tn epitopes on tumor cell lines. Mol. dynamics simulations indicated that in WT MGLshort, His259 mediates H bonds directly or engages the Tn-glycopeptide backbone through water mols. These bonds were lost in MGLshort H259T, thus explaining its lower binding affinity. Together, our results suggest that MGL not only connects to the Tn carbohydrate epitope, but also engages the underlying peptide via a secondary binding pocket within the MGL carbohydrate recognition domain contg. the His259 residue.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvVCjsL0%253D&md5=edab0eacc775c4e5b0eab684197e413a
    5. 5
      Zaal, A.; Li, R. J. E.; Lübbers, J.; Bruijns, S. C. M.; Kalay, H.; van Kooyk, Y.; van Vliet, S. J. Activation of the C-Type Lectin MGL by Terminal GalNAc Ligands Reduces the Glycolytic Activity of Human Dendritic Cells. Front. Immunol. 2020, 11, 305,  DOI: 10.3389/fimmu.2020.00305
      5
      Activation of the C-type lectin MGL by terminal GalNAc ligands reduces the glycolytic activity of human dendritic cells
      Zaal, Anouk; Eveline Li, R. J.; Luebbers, Joyce; Bruijns, Sven C. M.; Kalay, Hakan; van Kooyk, Yvette; van Vliet, Sandra J.
      Frontiers in Immunology (2020), 11 (), 305CODEN: FIRMCW; ISSN:1664-3224. (Frontiers Media S.A.)
      Many tumors display alterations in biosynthetic pathways of glycosylation, resulting in increased expression of specific tumor-assocd. glycan structures. Expression of these altered glycan structures is assocd. with metastasis and poor prognosis. Antigen presenting cells can recognize tumor-assocd. glycan structures, including the truncated O-glycan Tn antigen, via specific glycan receptors. Tn antigen-mediated activation of the C-type lectin MGL on dendritic cells induces regulatory T cells via the enhanced secretion of IL-10. Although these findings indicate that MGL engagement by glycan ligands can modulate immune responses, the impact of MGL ligation on dendritic cells is still not completely understood. Therefore, we employed RNA sequencing, GO term enrichment and pathway anal. on human monocyte-derived dendritic cells stimulated with two different MGL glycan ligands. Our analyses revealed a reduced expression of genes coding for key enzymes involved in the glycolysis pathway, TCA cycle, and oxidative phosphorylation. In concordance with this, extracellular flux anal. confirmed the decrease in glycolytic activity upon MGL triggering in human dendritic cells. To our knowledge, we are the first to report a diminished glycolytic activity of human dendritic cells upon C-type lectin stimulation. Overall, our findings highlight the impact of tumor-assocd. glycans on dendritic cell biol. and metab. and will increase our understanding on how glycans can shape immunity.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhvVarsr7E&md5=c34bc71b5c601783d553f49625e0451e
    6. 6
      Marcelo, F.; Garcia-Martin, F.; Matsushita, T.; Sardinha, J.; Coelho, H.; Oude-Vrielink, A.; Koller, C.; André, S.; Cabrita, E. J.; Gabius, H. J.; Nishimura, S. I.; Jiménez-Barbero, J.; Cañada, F. J. Delineating Binding Modes of Gal/GalNAc and Structural Elements of the Molecular Recognition of Tumor-Associated Mucin Glycopeptides by the Human Macrophage Galactose-Type Lectin. Chem.─Eur. J. 2014, 20, 16147– 16155,  DOI: 10.1002/chem.201404566
      6
      Delineating binding modes of Gal/GalNAc and structural elements of the molecular recognition of tumor-associated mucin glycopeptides by the human macrophage galactose-type lectin
      Marcelo, Filipa; Garcia-Martin, Fayna; Matsushita, Takahiko; Sardinha, Joao; Coelho, Helena; Oude-Vrielink, Anneloes; Koller, Christiane; Andre, Sabine; Cabrita, Eurico J.; Gabius, Hans-Joachim; Nishimura, Shin-ichiro; Jimenez-Barbero, Jesus; Canada, F. Javier
      Chemistry - A European Journal (2014), 20 (49), 16147-16155CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
      The human macrophage galactose-type lectin (MGL) is a key physiol. receptor for the carcinoma-assocd. Tn antigen (GalNAc-α-1-O-Ser/Thr) in mucins. NMR and modeling-based data on the mol. recognition features of synthetic Tn-bearing glycopeptides by MGL are presented. Cognate epitopes on the sugar and matching key amino acids involved in the interaction were identified by satn. transfer difference (STD) NMR spectroscopy. Only the amino acids close to the glycosylation site in the peptides are involved in lectin contact. Moreover, control expts. with non-glycosylated MUC1 peptides unequivocally showed that the sugar residue is essential for MGL binding, as is Ca2+. NMR data were complemented with mol. dynamics simulations and Corcema-ST to establish a 3D view on the mol. recognition process between Gal, GalNAc, and the Tn-presenting glycopeptides and MGL. Gal and GalNAc have a dual binding mode with opposite trend of the main interaction pattern and the differences in affinity can be explained by addnl. hydrogen bonds and CH-π contacts involving exclusively the NHAc moiety.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXitVCiurfN&md5=c30d5d96f7c5d59672d097293659fd42
    7. 7
      Mortezai, N.; Behnken, H. N.; Kurze, A. K.; Ludewig, P.; Buck, F.; Meyer, B.; Wagener, C. Tumor-Associated Neu5Ac-Tn and Neu5Gc-Tn Antigens Bind to C-Type Lectin CLEC10A (CD301, MGL). Glycobiology 2013, 23, 844– 852,  DOI: 10.1093/glycob/cwt021
      7
      Tumor-associated Neu5Ac-Tn and Neu5Gc-Tn antigens bind to C-type lectin CLEC10A (CD301, MGL)
      Mortezai, Naghmeh; Behnken, Henning N.; Kurze, Anna-Katharina; Ludewig, Peter; Buck, Friedrich; Meyer, Bernd; Wagener, Christoph
      Glycobiology (2013), 23 (7), 844-852CODEN: GLYCE3; ISSN:0959-6658. (Oxford University Press)
      In human tumors, glycoproteins often exhibit abnormal glycosylation patterns, e.g. certain Lewis structures, TF antigen, Tn antigen and/or their sialylated forms, creating addnl. binding sites for glycoreceptors. In the present study, we have analyzed the carbohydrate specificity of the C-type lectin CLEC10A using glycan profiling by ELISA (ELISA). In addn. to the known ligands, we show binding to two tumor-assocd. antigens, namely Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn, with an affinity of CLEC10A in the micromolar range. Detailed analyses of the glycan-lectin interactions were carried out by surface plasmon resonance (SPR) and satn. transfer difference (STD) NMR. CLEC10A binds Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn with dissocn. consts. of 297 and 80 μM, resp., as detd. by SPR. Comparison of the STD NMR (NMR) binding epitopes of Tn and Neu5Acα2,6-Tn revealed a const. binding mode of the N-acetylgalactosamine moiety. This finding is in good agreement with binding studies of CLEC10A transfectomas, which show a well-defined interaction of transmembrane CLEC10A with 6-sialylated-Tn structures. Since both Neu5Acα2,6-Tn and Neu5Gcα2,6-Tn together with the previously known Tn antigen are expressed in human tumors such as mammary carcinoma, the interaction with CLEC10A expressed by macrophages and dendritic cells could be of major functional significance in tumor progression.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXptFeruro%253D&md5=9c362e8087a962acf61ac6b5b8d959e2
    8. 8
      Bulteau, F.; Thépaut, M.; Henry, M.; Hurbin, A.; Vanwonterghem, L.; Vivès, C.; Le Roy, A.; Ebel, C.; Renaudet, O.; Fieschi, F.; Coll, J. L. Targeting Tn-Antigen-Positive Human Tumors with a Recombinant Human Macrophage Galactose C-Type Lectin. Mol. Pharm. 2022, 19, 235– 245,  DOI: 10.1021/acs.molpharmaceut.1c00744
      8
      Targeting Tn-Antigen-Positive Human Tumors with a Recombinant Human Macrophage Galactose C-Type Lectin
      Bulteau, Francois; Thepaut, Michel; Henry, Maxime; Hurbin, Amandine; Vanwonterghem, Laetitia; Vives, Corinne; Le Roy, Aline; Ebel, Christine; Renaudet, Olivier; Fieschi, Franck; Coll, Jean-Luc
      Molecular Pharmaceutics (2022), 19 (1), 235-245CODEN: MPOHBP; ISSN:1543-8384. (American Chemical Society)
      Alterations in glycosylation cause the emergence of tumor-assocd. carbohydrate antigens (TACAs) during tumorigenesis. Truncation of O-glycans reveals the Thomsen nouveau (Tn) antigen, an N-acetylgalactosamine (GalNAc) frequently attached to serine or threonine amino acids, that is accessible on the surface of cancer cells but not on healthy cells. Interestingly, GalNac can be recognized by macrophage galactose lectin (MGL), a type C lectin receptor expressed in immune cells. In this study, recombinant MGL fragments were tested in vitro for their cancer cell-targeting efficiency by flow cytometry and confocal microscopy and in vivo after administration of fluorescent MGL to tumor-bearing mice. Our results demonstrate the ability of MGL to target Tn-pos. human tumors without inducing toxicity. This outcome makes MGL, a fragment of a normal human protein, the first vector candidate for in vivo diagnosis and imaging of human tumors and, possibly, for therapeutic applications.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXislKks7rF&md5=53c31461c4343a34f3d81b59300b2609
    9. 9
      Cheever, M. A.; Allison, J. P.; Ferris, A. S.; Finn, O. J.; Hastings, B. M.; Hecht, T. T.; Mellman, I.; Prindiville, S. A.; Viner, J. L.; Weiner, L. M.; Matrisian, L. M. The Prioritization of Cancer Antigens: A National Cancer Institute Pilot Project for the Acceleration of Translational Research. Clin. Cancer Res. 2009, 15, 5323– 5337,  DOI: 10.1158/1078-0432.ccr-09-0737
      9
      The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research
      Cheever Martin A; Allison James P; Ferris Andrea S; Finn Olivera J; Hastings Benjamin M; Hecht Toby T; Mellman Ira; Prindiville Sheila A; Viner Jaye L; Weiner Louis M; Matrisian Lynn M
      Clinical cancer research : an official journal of the American Association for Cancer Research (2009), 15 (17), 5323-37 ISSN:.
      The purpose of the National Cancer Institute pilot project to prioritize cancer antigens was to develop a well-vetted, priority-ranked list of cancer vaccine target antigens based on predefined and preweighted objective criteria. An additional aim was for the National Cancer Institute to test a new approach for prioritizing translational research opportunities based on an analytic hierarchy process for dealing with complex decisions. Antigen prioritization involved developing a list of "ideal" cancer antigen criteria/characteristics, assigning relative weights to those criteria using pairwise comparisons, selecting 75 representative antigens for comparison and ranking, assembling information on the predefined criteria for the selected antigens, and ranking the antigens based on the predefined, preweighted criteria. Using the pairwise approach, the result of criteria weighting, in descending order, was as follows: (a) therapeutic function, (b) immunogenicity, (c) role of the antigen in oncogenicity, (d) specificity, (e) expression level and percent of antigen-positive cells, (f) stem cell expression, (g) number of patients with antigen-positive cancers, (h) number of antigenic epitopes, and (i) cellular location of antigen expression. None of the 75 antigens had all of the characteristics of the ideal cancer antigen. However, 46 were immunogenic in clinical trials and 20 of them had suggestive clinical efficacy in the "therapeutic function" category. These findings reflect the current status of the cancer vaccine field, highlight the possibility that additional organized efforts and funding would accelerate the development of therapeutically effective cancer vaccines, and accentuate the need for prioritization.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BD1Mrpslamsw%253D%253D&md5=bf8652477b44e40ca6da6b83f8ec471d
    10. 10
      Nath, S.; Mukherjee, P. MUC1: A Multifaceted Oncoprotein with a Key Role in Cancer Progression. Trends Mol. Med. 2014, 20, 332– 342,  DOI: 10.1016/j.molmed.2014.02.007
      10
      MUC1: a multifaceted oncoprotein with a key role in cancer progression
      Nath, Sritama; Mukherjee, Pinku
      Trends in Molecular Medicine (2014), 20 (6), 332-342CODEN: TMMRCY; ISSN:1471-4914. (Elsevier Ltd.)
      A review. The transmembrane glycoprotein Mucin 1 (MUC1) is aberrantly glycosylated and overexpressed in a variety of epithelial cancers, and plays a crucial role in progression of the disease. Tumor-assocd. MUC1 differs from the MUC1 expressed in normal cells with regard to its biochem. features, cellular distribution, and function. In cancer cells, MUC1 participates in intracellular signal transduction pathways and regulates the expression of its target genes at both the transcriptional and post-transcriptional levels. This review highlights the structural and functional differences that exist between normal and tumor-assocd. MUC1. We also discuss the recent advances made in the use of MUC1 as a biomarker and therapeutic target for cancer.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXksleqs7w%253D&md5=73e37d4a1e83b57e90078c2b4a4afbc0
    11. 11
      Springer, G. F. Immunoreactive T and Tn Epitopes in Cancer Diagnosis, Prognosis, and Immunotherapy. J. Mol. Med. 1997, 75, 594– 602,  DOI: 10.1007/s001090050144
      11
      Immunoreactive T and Tn epitopes in cancer diagnosis, prognosis, and immunotherapy
      Springer, Georg F.
      Journal of Molecular Medicine (Berlin) (1997), 75 (8), 594-602CODEN: JMLME8; ISSN:0946-2716. (Springer)
      A review with 74 refs. Aberrant glycosylation is a hallmark of cancer cells. The blood group precursors T (Thomsen-Friedenreich) and Tn epitopes are shielded in healthy and benign-diseased tissues but uncovered in approx. 90% of carcinomas. T and Tn glycoproteins are specific, autoimmunogenic pancarcinoma antigens. These antigens may also be found in neoplastic blood cells (and on LTV-II infected T lymphocytes). Fundamental chem. and phys. aspects of these glycoproteins of primary carcinomas are discussed first. Tn and T epitopes are cell and tissue adhesion mols., essential in invasion by and metastasis of carcinoma which includes adherent and proliferative phases. These properties are then delineated next, followed by consideration of pathophysiol. and clin. aspects of these antigens. Immunohistochem. studies of the extent of Tn antigen expression in primary breast carcinoma demonstrate its highly significant correlation with clinicopathol. tumor stages, and hence its value as a reliable prognosticator. On the other hand, there is no significant, prognostically useful assocn. between T antigen expression and clin. disease course. Everyone has "preexisting" anticarcinoma anti-Tn and anti-T antibodies, induced predominantly by the intestinal flora, while cellular immune responses to T and Tn epitopes are evoked only by carcinomas and some lymphomas. Carcinoma dedifferentiation leading to predominance of Tn over T epitopes is described, as is the role of Tn and T epitopes in very early, including preclin., carcinoma detection. The highest sensitivities in carcinoma detection are for preclin. and the earliest clin. stages. Obviously, preclin. carcinoma detection is of practical importance. T/anti-T tests detected preclin. carcinoma in 77% of 48 patients long (‾ 6 yr) before their biopsy/X-ray results became pos. There were no false predictions of carcinoma in 38 control persons with benign diseases (observation av. 4.8 yr). These findings open a novel window for both curative approaches and pathophysiol. studies. The autoimmunogenicity of carcinoma T/Tn antigen led us more than two decades ago to begin intradermal vaccination of patients with advanced breast carcinoma of stages IV-IIb, predominately after modified radical mastectomy and sometimes lumpectomy plus axillary dissection always followed by adjuvant radio/chemotherapy. The vaccine consists of human group O red blood cell membrane derived, HLA-free T/Tn antigen contg. as adjuvant Ca3(PO4)2 plus a trace of phosphoglycolipid A hyperantigen, i.e., S. typhi vaccine (USP), which itself has T and Tn specificities. Our efforts have now for up to 20 yr remained successful in a large majority of the 32 patients. All 32 patients survived at least 5 yr; 10-yr survival was statistically highly significantly improved (5-yr survival: P<1×10-7; 10-yr survival: P<1×10-5) compared to statistics of the United States National Cancer Institute. Because these vaccinations are successful, their extension to large populations with major types of carcinomas should be considered, and even immunol. carcinoma prevention may be contemplated.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXlvVKjtL8%253D&md5=0d81fbc8c7a7835e58872c3c707a2115
    12. 12
      Beckwith, D. M.; Cudic, M. Tumor-Associated O-Glycans of MUC1: Carriers of the Glyco-Code and Targets for Cancer Vaccine Design. Semin Immunol. 2020, 47, 101389,  DOI: 10.1016/j.smim.2020.101389
      12
      Tumor-associated O-glycans of MUC1: Carriers of the glyco-code and targets for cancer vaccine design
      Beckwith, Donella M.; Cudic, Mare
      Seminars in Immunology (2020), 47 (), 101389CODEN: SEIME2; ISSN:1044-5323. (Elsevier Ltd.)
      A review. The transformation from normal to malignant phenotype in human cancers is assocd. with aberrant cell-surface glycosylation. It has frequently been reported that MUC1, the heavily glycosylated cell-surface mucin, is altered in both, expression and glycosylation pattern, in human carcinomas of the epithelium. The presence of incomplete or truncated glycan structures, often capped by sialic acid, commonly known as tumor-assocd. carbohydrate antigens (TACAs), play a key role in tumor initiation, progression, and metastasis. Accumulating evidence suggests that expression of TACAs is assocd. with tumor escape from immune defenses. In this report, we will give an overview of the oncogenic functions of MUC1 that are exerted through TACA interactions with endogenous carbohydrate-binding proteins (lectins). These interactions often lead to creation of a pro-tumor microenvironment, favoring tumor progression and metastasis, and tumor evasion. In addn., we will describe current efforts in the design of cancer vaccines with special emphasis on synthetic MUC1 glycopeptide vaccines. Anal. of the key factors that govern structure-based design of immunogenic MUC1 glycopeptide epitopes are described. The role of TACA type, position, and d. on obsd. humoral and cellular immune responses is evaluated.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXlslemsQ%253D%253D&md5=42abcf8643445549c0fe38087b7b7640
    13. 13
      Gaidzik, N.; Westerlind, U.; Kunz, H. The Development of Synthetic Antitumour Vaccines from Mucin Glycopeptide Antigens. Chem. Soc. Rev. 2013, 42, 4421– 4442,  DOI: 10.1039/c3cs35470a
      13
      The development of synthetic antitumor vaccines from mucin glycopeptide antigens
      Gaidzik, Nikola; Westerlind, Ulrika; Kunz, Horst
      Chemical Society Reviews (2013), 42 (10), 4421-4442CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
      A review. Based on important cell-biol. and biochem. results concerning the structural difference between membrane glycoproteins of normal epithelial cells and epithelial tumor cells, tumor-assocd. glycopeptide antigens were chem. synthesized and structurally confirmed. Glycopeptide structures of the tandem repeat sequence of mucin MUC1 of epithelial tumor cells constitute the most promising tumor-assocd. antigens. In order to generate a sufficient immunogenicity of these endogenous structures, usually tolerated by the immune system, these synthetic glycopeptide antigens were conjugated to immune stimulating components: in fully synthetic 2-component vaccines either with T-cell peptide epitopes or with Toll-like receptor2 lipopeptide ligands or in 3-component vaccines with both these stimulants. Alternatively, the synthetic glycopeptide antigens were coupled to immune stimulating carrier proteins. In particular, MUC1 glycopeptide conjugates with Tetanus toxoid proved to be efficient vaccines inducing very strong immune responses in mice. The antibodies elicited with the fully synthetic vaccines showed selective recognition of the tumor-assocd. glycopeptides as was shown by neutralization and micro-array binding expts. After booster immunizations, most of the immune responses showed the installation of an immunol. memory. Immunization with fully synthetic 3-component vaccines induced immune reactions with therapeutic effects in terms of redn. of the tumor burden in mice or in killing of tumor cells in culture, while MUC1 glycopeptide-Tetanus toxoid vaccines elicited antibodies in mice which recognized tumor cells in human tumor tissues. The results achieved so far are considered to be promising for the development of an active immunization against tumors.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXms1Gitb0%253D&md5=038515ff3ac8ec3851429dbe8866a781
    14. 14
      Stergiou, N.; Urschbach, M.; Gabba, A.; Schmitt, E.; Kunz, H.; Besenius, P. The Development of Vaccines from Synthetic Tumor-Associated Mucin Glycopeptides and Their Glycosylation-Dependent Immune Response. Chem. Rec. 2021, 21, 3313– 3331,  DOI: 10.1002/tcr.202100182
      14
      The Development of Vaccines from Synthetic Tumor-Associated Mucin Glycopeptides and their Glycosylation-Dependent Immune Response
      Stergiou, Natascha; Urschbach, Moritz; Gabba, Adele; Schmitt, Edgar; Kunz, Horst; Besenius, Pol
      Chemical Record (2021), 21 (11), 3313-3331CODEN: CRHEAK; ISSN:1528-0691. (Wiley-VCH Verlag GmbH & Co. KGaA)
      A review. Tumor-assocd. carbohydrate antigens are overexpressed as altered-self in most common epithelial cancers. Their glycosylation patterns differ from those of healthy cells, functioning as an ID for cancer cells. Scientists have been developing anti-cancer vaccines based on mucin glycopeptides, yet the interplay of delivery system, adjuvant and tumor assocd. MUC epitopes in the induced immune response is not well understood. The current state of the art suggests that the identity, abundancy and location of the glycans on the MUC backbone are all key parameters in the cellular and humoral response. This review shares lessons learned by us in over two decades of research in glycopeptide vaccines. By bridging synthetic chem. and immunol., we discuss efforts in designing synthetic MUC1/4/16 vaccines and focus on the role of glycosylation patterns. We provide a brief introduction into the mechanisms of the immune system and aim to promote the development of cancer subunit vaccines.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXisFOiurzI&md5=83bfea0e1d565dc62594bc6566d06ae5
    15. 15
      Rowse, G. J.; Tempero, R. M.; VanLith, M. L.; Hollingsworth, M. A.; Gendler, S. J. Tolerance and Immunity to MUC1 in a Human MUC1 Transgenic Murine Model. Cancer Res. 1998, 58, 315– 321
      15
      Tolerance and immunity to MUC1 in a human MUC1 transgenic murine model
      Rowse, Gerald J.; Tempero, Richard M.; VanLith, Michelle L.; Hollingsworth, Michael A.; Gendler, Sandra J.
      Cancer Research (1998), 58 (2), 315-321CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
      The human epithelial mucin, MUC1, is a large transmembrane glycoprotein that is expressed on most simple epithelia. It is overexpressed and aberrantly glycosylated on many human epithelial tumors, including more than 90% of human breast cancers. MUC1 is of interest as an immunotherapy target because patients with breast, ovarian, and pancreatic cancers have T lymphocytes in their tumor-draining lymph nodes that can be induced to recognize and lyse MUC1-expressing tumor cells. We have produced a transgenic mouse model that expresses the human MUC1 mol. on an inbred C57Bl/6 background to investigate the effect of endogenous expression of MUC1 on the ability of mice to generate anti-tumor immunity to MUC1-expressing tumors. Transgenic mice expressed the human transgene in a pattern and level consistent with that obsd. in humans. Transgenic mice were tolerant to stimulation by MUC1 as evidenced by the ability of MUC1-expressing tumor cells to grow in these mice, whereas MUC1-expressing cells were eliminated from wild-type mice. Moreover, transgenic mice immunized with MUC1 peptides failed to exhibit Ig class switching to the IgG subtypes. These data suggest that endogenous expression of MUC1 protein by MUC1 transgenic mice induces T-cell tolerance to stimulation by MUC1. The transgenic mice will provide a useful model to investigate the mechanisms that regulate immunol. tolerance to tumor antigens and will facilitate the investigation of anti-MUC1 immunotherapy formulations.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1cXntF2gtQ%253D%253D&md5=4dd48bf8e125ca46d65dca946d6b3171
    16. 16
      Brown, G. D.; Willment, J. A.; Whitehead, L. C-type lectins in immunity and homeostasis. Nat. Rev. Immunol. 2018, 18, 374– 389,  DOI: 10.1038/s41577-018-0004-8
      16
      C-type lectins in immunity and homeostasis
      Brown, Gordon D.; Willment, Janet A.; Whitehead, Lauren
      Nature Reviews Immunology (2018), 18 (6), 374-389CODEN: NRIABX; ISSN:1474-1733. (Nature Research)
      A review. The C-type lectins are a superfamily of proteins that recognize a broad repertoire of ligands and that regulate a diverse range of physiol. functions. Most research attention has focused on the ability of C-type lectins to function in innate and adaptive antimicrobial immune responses, but these proteins are increasingly being recognized to have a major role in autoimmune diseases and to contribute to many other aspects of multicellular existence. Defects in these mols. lead to developmental and physiol. abnormalities, as well as altered susceptibility to infectious and non-infectious diseases. In this Review, we present an overview of the roles of C-type lectins in immunity and homeostasis, with an emphasis on the most exciting recent discoveries.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhtFWqs7jI&md5=9da9ec71ab6fb3262f297cff160aa72d
    17. 17
      Ilarregui, J. M.; Kooij, G.; Rodríguez, E.; Van Der Pol, S. M. A.; Koning, N.; Kalay, H.; Van Der Horst, J. C.; Van Vliet, S. J.; García-Vallejo, J. J.; De Vries, H. E.; Van Kooyk, Y. Macrophage Galactose-Type Lectin (MGL) Is Induced on M2 Microglia and Participates in the Resolution Phase of Autoimmune Neuroinflammation. J. Neuroinflammation 2019, 16, 130– 214,  DOI: 10.1186/s12974-019-1522-4
      17
      Macrophage galactose-type lectin (MGL) is induced on M2 microglia and participates in the resolution phase of autoimmune neuroinflammation
      Ilarregui Juan M; Rodriguez Ernesto; Koning Nathalie; Kalay Hakan; van der Horst Joost C; van Vliet Sandra J; Garcia-Vallejo Juan J; van Kooyk Yvette; Kooij Gijs; van der Pol Susanne M A; de Vries Helga E
      Journal of neuroinflammation (2019), 16 (1), 130 ISSN:.
      BACKGROUND: Multiple sclerosis (MS) involves a misdirected immune attack against myelin in the brain and spinal cord, leading to profound neuroinflammation and neurodegeneration. While the mechanisms of disease pathogenesis have been widely studied, the suppression mechanisms that lead to the resolution of the autoimmune response are still poorly understood. Here, we investigated the role of the C-type lectin receptor macrophage galactose-type lectin (MGL), usually expressed on tolerogenic antigen-presenting cells (APCs), as a negative regulator of autoimmune-driven neuroinflammation. METHODS: We used in silico, immunohistochemical, immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR) and flow cytometry analysis to explore the expression and functionality of MGL in human macrophages and microglia, as well as in MS post-mortem tissue. In vitro, we studied the capacity of MGL to mediate apoptosis of experimental autoimmune encephalomyelitis (EAE)-derived T cells and mouse CD4(+) T cells. Finally, we evaluated in vivo and ex vivo the immunomodulatory potential of MGL in EAE. RESULTS: MGL plays a critical role in the resolution phase of EAE as MGL1-deficient (Clec10a(-/-)) mice showed a similar day of onset but experienced a higher clinical score to that of WT littermates. We demonstrate that the mouse ortholog MGL1 induces apoptosis of autoreactive T cells and diminishes the expression of pro-inflammatory cytokines and inflammatory autoantibodies. Moreover, we show that MGL1 but not MGL2 induces apoptosis of activated mouse CD4(+) T cells in vitro. In human settings, we show that MGL expression is increased in active MS lesions and on alternatively activated microglia and macrophages which, in turn, induces the secretion of the immunoregulatory cytokine IL-10, underscoring the clinical relevance of this lectin. CONCLUSIONS: Our results show a new role of MGL-expressing APCs as an anti-inflammatory mechanism in autoimmune neuroinflammation by dampening pathogenic T and B cell responses, uncovering a novel clue for neuroprotective therapeutic strategies with relevance for in MS clinical applications.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3Mzhs1Gqtg%253D%253D&md5=431c7d27f54f82c5dff177e464034b71
    18. 18
      Costa, M.; da Costa, V.; Lores, P.; Landeira, M.; Rodríguez-Zraquia, S. A.; Festari, M. F.; Freire, T. Macrophage Gal/GalNAc Lectin 2 (MGL2)+ Peritoneal Antigen Presenting Cells during Fasciola Hepatica Infection Are Essential for Regulatory T Cell Induction. Sci. Rep. 2022, 12, 17661– 17712,  DOI: 10.1038/s41598-022-21520-w
      18
      Macrophage Gal/GalNAc lectin 2 (MGL2)+ peritoneal antigen presenting cells during Fasciola hepatica infection are essential for regulatory T cell induction
      Costa, Monique; da Costa, Valeria; Lores, Pablo; Landeira, Mercedes; Rodriguez-Zraquia, Santiago A.; Festari, Maria Florencia; Freire, Teresa
      Scientific Reports (2022), 12 (1), 17661CODEN: SRCEC3; ISSN:2045-2322. (Nature Portfolio)
      Fasciola hepatica, one of the agents that causes fasciolosis, modulates the host immune system to allow parasite survival in the host. F. hepatica expresses carbohydrate-contg. glycoconjugates that are decoded by C-type lectin receptors, such as Dectin-1, mannose receptor, DC-SIGN and MGL, that are mainly present on myeloid antigen presenting cells (APCs) and can mediate immunoregulatory properties on T cells. In particular, Macrophage Gal/GalNAc lectin 2 (MGL2) expands modified Th2 immune responses, while suppressing Th1 polarization, upon recognition of GalNAc-glycosylated parasite components. In this study, by using MGL2-DTR transgenic mice that encode human diphtheria toxin receptor in MGL2+ cells, we demonstrate the role of peritoneal APCs during F. hepatica infection in favoring parasite survival. This process might be mediated by the induction of splenic Tregs in vivo, since the depletion of MGL2+ cells conferred mice with partial resistance to the infection and abrogated the increase of CD4+/CD25+ FoxP3+ Tregs induced by the parasite. Therefore, MGL2+ cells are crit. determinants of F. hepatica infection and could constitute immune checkpoints to control parasite infection.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XislaksLzP&md5=8bf1f3ef110b3c9652c20404bec5ca1d
    19. 19
      Pace, A.; Scirocchi, F.; Napoletano, C.; Zizzari, I. G.; D’angelo, L.; Santoro, A.; Nuti, M.; Rahimi, H.; Rughetti, A. Glycan-Lectin Interactions as Novel Immunosuppression Drivers in Glioblastoma. Int. J. Mol. Sci. 2022, 23, 6312,  DOI: 10.3390/ijms23116312
      19
      Glycan-Lectin Interactions as Novel Immunosuppression Drivers in Glioblastoma
      Pace, Angelica; Scirocchi, Fabio; Napoletano, Chiara; Zizzari, Ilaria Grazia; D'Angelo, Luca; Santoro, Antonio; Nuti, Marianna; Rahimi, Hassan; Rughetti, Aurelia
      International Journal of Molecular Sciences (2022), 23 (11), 6312CODEN: IJMCFK; ISSN:1422-0067. (MDPI AG)
      A review. Despite diagnostic and therapeutic improvements, glioblastoma (GB) remains one of the most threatening brain tumor in adults, underlining the urgent need of new therapeutic targets. Lectins are glycan-binding proteins that regulate several biol. processes through the recognition of specific sugar motifs. Lectins and their ligands are found on immune cells, endothelial cells and, also, tumor cells, pointing out a strong correlation among immunity, tumor microenvironment and vascularization. In GB, altered glycans and lectins contribute to tumor progression and immune evasion, shaping the tumor-immune landscape promoting immunosuppressive cell subsets, such as myeloid-derived suppressor cells (MDSCs) and M2-macrophages, and affecting immunoeffector populations, such as CD8+ T cells and dendritic cells (DCs). Here, we discuss the latest knowledge on the immune cells, immune related lectin receptors (C-type lectins, Siglecs, galectins) and changes in glycosylation that are involved in immunosuppressive mechanisms in GB, highlighting their interest as possible novel therapeutical targets.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XhsFWru73N&md5=4b611e66666a11371dab01a910cc502d
    20. 20
      da Costa, V.; van Vliet, S. J.; Carasi, P.; Frigerio, S.; García, P. A.; Croci, D. O.; Festari, M. F.; Costa, M.; Landeira, M.; Rodríguez-Zraquia, S. A.; Cagnoni, A. J.; Cutine, A. M.; Rabinovich, G. A.; Osinaga, E.; Mariño, K. V.; Freire, T. The Tn Antigen Promotes Lung Tumor Growth by Fostering Immunosuppression and Angiogenesis via Interaction with Macrophage Galactose-Type Lectin 2 (MGL2). Cancer Lett. 2021, 518, 72– 81,  DOI: 10.1016/j.canlet.2021.06.012
      20
      The Tn antigen promotes lung tumor growth by fostering immunosuppression and angiogenesis via interaction with Macrophage Galactose-type lectin 2 (MGL2)
      da Costa, Valeria; van Vliet, Sandra J.; Carasi, Paula; Frigerio, Sofia; Garcia, Pablo A.; Croci, Diego O.; Festari, Maria Florencia; Costa, Monique; Landeira, Mercedes; Rodriguez-Zraquia, Santiago A.; Cagnoni, Alejandro J.; Cutine, Anabela M.; Rabinovich, Gabriel A.; Osinaga, Eduardo; Marino, Karina V.; Freire, Teresa
      Cancer Letters (New York, NY, United States) (2021), 518 (), 72-81CODEN: CALEDQ; ISSN:0304-3835. (Elsevier Inc.)
      Tn is a tumor-assocd. carbohydrate antigen that constitutes both a diagnostic tool and an immunotherapeutic target. It originates from interruption of the mucin O-glycosylation pathway through defects involving, at least in part, alterations in core-1 synthase activity, which is highly dependent on Cosmc, a folding chaperone. Tn antigen is recognized by the Macrophage Galactose-type Lectin (MGL), a C-type lectin receptor present on dendritic cells and macrophages. Specific interactions between Tn and MGL shape anti-tumoral immune responses by regulating several innate and adaptive immune cell programs. In this work, we generated and characterized a variant of the lung cancer murine cell line LL/2 that expresses Tn by mutation of the Cosmc chaperone gene (Tn+ LL/2). We confirmed Tn expression by lectin glycophenotyping and specific anti-Tn antibodies, verified abrogation of T-synthase activity in these cells, and confirmed its recognition by the murine MGL2 receptor. Interestingly, Tn+ LL/2 cells were more aggressive in vivo, resulting in larger and highly vascularized tumors than those generated from wild type Tn- LL/2 cells. In addn., Tn+ tumors exhibited an increase in CD11c+ F4/80+ cells with high expression of MGL2, together with an augmented expression of IL-10 in infiltrating CD4+ and CD8+ T cells. Importantly, this immunosuppressive microenvironment was dependent on the presence of MGL2+ cells, since depletion of these cells abrogated tumor growth, vascularization and recruitment of IL-10+ T cells. Altogether, our results suggest that expression of Tn in tumor cells and its interaction with MGL2-expressing CD11c+F4/80+ cells promote immunosuppression and angiogenesis, thus favoring tumor progression.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhtlKrtbfP&md5=3dcc08a15f2b731d5a4c66d19ae857a1
    21. 21
      Freire, T.; Lo-Man, R.; Bay, S.; Leclerc, C. Tn Glycosylation of the MUC6 Protein Modulates Its Immunogenicity and Promotes the Induction of Th17-Biased T Cell Responses. J. Biol. Chem. 2011, 286, 7797– 7811,  DOI: 10.1074/jbc.m110.209742
      21
      Tn Glycosylation of the MUC6 Protein Modulates its Immunogenicity and Promotes the Induction of Th17-biased T Cell Responses
      Freire, Teresa; Lo-Man, Richard; Bay, Sylvie; Leclerc, Claude
      Journal of Biological Chemistry (2011), 286 (10), 7797-7811CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
      The Tn antigen (α-GalNAc-O-Ser/Thr) is one of the most specific human cancer-assocd. structures. This antigen, together with mucins, the major carriers of O-glycosylated tumor antigens in adenocarcinomas, are being evaluated as anti-cancer immunotherapeutic targets. In particular, the MUC6 protein, which is normally expressed only in gastric tissues, has been detected in intestinal, pulmonary, colorectal, and breast carcinomas. To develop anti-cancer vaccines based on the Tn antigen, the authors produced MUC6 proteins with different Tn d. by using mixts. of recombinant ppGalNAc-T1, -T2, and -T7. The obtained glycoproteins were characterized and analyzed for their immunol. properties, as compared with the non-glycosylated MUC6. The authors show that these various MUC6:Tn glycoproteins were well recognized by both MUC6 and Tn-specific antibodies. However, Tn glycosylation of the MUC6 protein strongly affected their immunogenicity by partially abrogating Th1 cell responses, and promoting IL-17 responses. Moreover, the non-glycosylated MUC6 was more efficiently presented than MUC6:Tn glycoproteins to specific T CD4+ hybridomas, suggesting that Tn glycosylation may affect MUC6 processing or MHC binding of the processed peptides. In conclusion, the results indicate that Tn glycosylation of the MUC6 protein strongly affects its B and T cell immunogenicity, and might favor immune escape of tumor cells.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXislylsL8%253D&md5=fd44b4f44acf8d5e5ac975a79b421a65
    22. 22
      Freire, T.; Zhang, X.; Dériaud, E.; Ganneau, C.; Vichier-Guerre, S.; Azria, E.; Launay, O.; Lo-Man, R.; Bay, S.; Leclerc, C. Glycosidic Tn-Based Vaccines Targeting Dermal Dendritic Cells Favor Germinal Center B-Cell Development and Potent Antibody Response in the Absence of Adjuvant. Blood 2010, 116, 3526– 3536,  DOI: 10.1182/blood-2010-04-279133
      22
      Glycosidic Tn-based vaccines targeting dermal dendritic cells favor germinal center B-cell development and potent antibody response in the absence of adjuvant
      Freire, Teresa; Zhang, Xiaoming; Deriaud, Edith; Ganneau, Christelle; Vichier-Guerre, Sophie; Azria, Elie; Launay, Odile; Lo-Man, Richard; Bay, Sylvie; Leclerc, Claude
      Blood (2010), 116 (18), 3526-3536CODEN: BLOOAW; ISSN:0006-4971. (American Society of Hematology)
      In vivo targeting of C-type lectin receptors is an effective strategy for increasing antigen uptake and presentation by dendritic cells (DCs). To induce efficient immune response, glycosylated tumor-assocd. Tn antigens were used to target DCs through binding to macrophage galactose-type lectin (MGL). The capacity of Tn-glycosylated antigens-and the multiple antigenic glycopeptide Tn3 therapeutic candidate vaccine-to target mouse and human MGL+ DCs are demonstrated, esp. regarding dermal DCs. In mice, MGL+ CD103- dermal DCs efficiently captured and processed glycosylated Tn antigen in vivo, inducing a potent major histocompatibility complex (MHC) class II-restricted T-cell response. Intradermal immunization with Tn-glycopeptides induced high levels of Th2 cytokines-even in the presence of unmethylated cytosine-phosphate-guanosine-and was assocd. with increased expansion of the germinal center B-cell population. Therefore, MGL acts as an efficient endocytic antigen receptor on dermal DCs in vivo, able to prime Tn-specific T- and B-cell responses. Moreover, even in the absence of adjuvant, immunization with this glycosidic Tn-based vaccine induced high levels of anti-Tn antibody responses, recognizing human tumor cells. In vivo DC-targeting strategies, based on Tn-MGL interactions, constitute a promising strategy for enhancing antigen presentation and inducing potent antibody response.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhsVKksb7J&md5=0da063b69b6371bdb6d5f4687b13ce49
    23. 23
      Rosenbaum, P.; Artaud, C.; Bay, S.; Ganneau, C.; Campone, M.; Delaloge, S.; Gourmelon, C.; Loirat, D.; Medioni, J.; Pein, F.; Sablin, M. P.; Tredan, O.; Varga, A.; Leclerc, C. The Fully Synthetic Glycopeptide MAG-Tn3 Therapeutic Vaccine Induces Tumor-Specific Cytotoxic Antibodies in Breast Cancer Patients. Cancer Immunol. Immunother. 2020, 69, 703– 716,  DOI: 10.1007/s00262-020-02503-0
      23
      The fully synthetic glycopeptide MAG-Tn3 therapeutic vaccine induces tumor-specific cytotoxic antibodies in breast cancer patients
      Rosenbaum, Pierre; Artaud, Cecile; Bay, Sylvie; Ganneau, Christelle; Campone, Mario; Delaloge, Suzette; Gourmelon, Carole; Loirat, Delphine; Medioni, Jacques; Pein, Francois; Sablin, Marie-Paule; Tredan, Olivier; Varga, Andrea; Leclerc, Claude
      Cancer Immunology Immunotherapy (2020), 69 (5), 703-716CODEN: CIIMDN; ISSN:0340-7004. (Springer)
      Cancer is one of the main causes of mortality worldwide and a major public health concern. Among various strategies, therapeutic vaccines have been developed to stimulate anti-tumoral immune responses. However, in spite of extensive studies, this approach suffers from a lack of efficacy. We designed the MAG-Tn3 vaccine, aiming to induce antibody responses against Tn, a tumor-assocd. carbohydrate antigen. The fully synthetic glycopeptide vaccine MAG-Tn3 is composed of four arms built on three adjacent Tn moieties assocd. with the tetanus toxin-derived peptide TT830-844 CD4+ T-cell epitope. This promiscuous CD4+ T-cell epitope can bind to a wide range of HLA-DRB mols. and is thus expected to activate CD4+ T-cell responses in a large part of the human population. The MAG-Tn3 vaccine was formulated with the GSK-proprietary immunostimulant AS15, composed of CpG7909, MPL, and QS21, which has been shown to stimulate both innate and humoral responses, in addn. to being well tolerated. The first results of phase I clin. trial demonstrated that all vaccinated patients developed high levels of Tn-specific antibodies. Moreover, these antibodies specifically recognized Tn-expressing human tumor cells and killed them through a complement-dependent cytotoxicity mechanism. Overall, this study establishes, for the first time, the capacity of a fully synthetic glycopeptide cancer vaccine to induce specific immune responses in humans.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjtlajur0%253D&md5=446ca7adcafd30823c3a3158da8c9b2e
    24. 24
      Laubreton, D.; Bay, S.; Sedlik, C.; Artaud, C.; Ganneau, C.; Dériaud, E.; Viel, S.; Puaux, A. L.; Amigorena, S.; Gérard, C.; Lo-Man, R.; Leclerc, C. The Fully Synthetic MAG-Tn3 Therapeutic Vaccine Containing the Tetanus Toxoid-Derived TT830-844 Universal Epitope Provides Anti-Tumor Immunity. Cancer Immunol. Immunother. 2016, 65, 315– 325,  DOI: 10.1007/s00262-016-1802-0
      24
      The fully synthetic MAG-Tn3 therapeutic vaccine containing the tetanus toxoid-derived TT830-844 universal epitope provides anti-tumor immunity
      Laubreton, Daphne; Bay, Sylvie; Sedlik, Christine; Artaud, Cecile; Ganneau, Christelle; Deriaud, Edith; Viel, Sophie; Puaux, Anne-Laure; Amigorena, Sebastian; Gerard, Catherine; Lo-Man, Richard; Leclerc, Claude
      Cancer Immunology Immunotherapy (2016), 65 (3), 315-325CODEN: CIIMDN; ISSN:0340-7004. (Springer)
      Malignant transformations are often assocd. with aberrant glycosylation processes that lead to the expression of new carbohydrate antigens at the surface of tumor cells. Of these carbohydrate antigens, the Tn antigen is particularly highly expressed in many carcinomas, esp. in breast carcinoma. We designed MAG-Tn3, a fully synthetic vaccine based on three consecutive Tn moieties that are O-linked to a CD4+ T cell epitope, to induce anti-Tn antibody responses that could be helpful for therapeutic vaccination against cancer. To ensure broad coverage within the human population, the tetanus toxoid-derived peptide TT830-844 was selected as a T-helper epitope because it can bind to various HLA-DRB mols. We showed that the MAG-Tn3 vaccine, which was formulated with the GSK proprietary immunostimulant AS15 and designed for human cancer therapy, is able to induce an anti-Tn antibody response in mice of various H-2 haplotypes, and this response correlates with the ability to induce a specific T cell response against the TT830-844 peptide. The universality of the TT830-844 peptide was extended to new H-2 and HLA-DRB mols. that were capable of binding this T cell epitope. Finally, the MAG-Tn3 vaccine was able to induce anti-Tn antibody responses in cynomolgus monkeys, which targeted Tn-expressing tumor cells and mediated tumor cell death both in vitro and in vivo. Thus, MAG-Tn3 is a highly promising anticancer vaccine that is currently under evaluation in a phase I clin. trial.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XisFSmsr4%253D&md5=913211d77c205902ac2a681ce07698a5
    25. 25
      Eggink, L. L.; Roby, K. F.; Cote, R.; Kenneth Hoober, J. An Innovative Immunotherapeutic Strategy for Ovarian Cancer: CLEC10A and Glycomimetic Peptides. J. Immunother. Cancer 2018, 6, 28,  DOI: 10.1186/s40425-018-0339-5
      25
      An innovative immunotherapeutic strategy for ovarian cancer: CLEC10A and glycomimetic peptides
      Eggink Laura L; Cote Robert; Kenneth Hoober J; Roby Katherine F
      Journal for immunotherapy of cancer (2018), 6 (1), 28 ISSN:.
      BACKGROUND: Receptors specific for the sugar N-acetylgalactosamine (GalNAc) include the human type II, C-type lectin receptor macrophage galactose-type lectin/C-type lectin receptor family member 10A (MGL/CLEC10A/CD301) that is expressed prominently by human peripheral immature dendritic cells, dendritic cells in the skin, alternatively-activated (M2a) macrophages, and to lesser extents by several other types of tissues. CLEC10A is an endocytic receptor on antigen-presenting cells and has been proposed to play an important role in maturation of dendritic cells and initiation of an immune response. In this study, we asked whether a peptide that binds in the GalNAc-binding site of CLEC10A would serve as an effective tool to activate an immune response against ovarian cancer. METHODS: A 12-mer sequence emerged from a screen of a phage display library with a GalNAc-specific lectin. The peptide, designated svL4, and a shorter peptide consisting of the C-terminal 6 amino acids, designated sv6D, were synthesized as tetravalent structures based on a tri-lysine core. In silico and in vitro binding assays were developed to evaluate binding of the peptides to GalNAc-specific receptors. Endotoxin-negative peptide solutions were administered by subcutaneous injection and biological activity of the peptides was determined by secretion of cytokines and the response of peritoneal immune cells in mice. Anti-cancer activity was studied in a murine model of ovarian cancer. RESULTS: The peptides bound to recombinant human CLEC10A with high avidity, with half-maximal binding in the low nanomolar range. Binding to the receptor was Ca(2+)-dependent. Subcutaneous injection of low doses of peptides into mice on alternate days resulted in several-fold expansion of populations of mature immune cells within the peritoneal cavity. Peptide sv6D effectively suppressed development of ascites in a murine ovarian cancer model as a monotherapy and in combination with the chemotherapeutic drug paclitaxel or the immunotherapeutic antibody against the receptor PD-1. Toxicity, including antigenicity and release of cytotoxic levels of cytokines, was not observed. CONCLUSION: sv6D is a functional ligand for CLEC10A and induces maturation of immune cells in the peritoneal cavity. The peptide caused a highly significant extension of survival of mice with implanted ovarian cancer cells with a favorable toxicity and non-antigenic profile.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1MjjslCrtA%253D%253D&md5=cd7dab036f346f98d7bccb199c31e183
    26. 26
      Heger, L.; Balk, S.; Lühr, J. J.; Heidkamp, G. F.; Lehmann, C. H. K.; Hatscher, L.; Purbojo, A.; Hartmann, A.; Garcia-Martin, F.; Nishimura, S. I.; Cesnjevar, R.; Nimmerjahn, F.; Dudziak, D. CLEC10A Is a Specific Marker for Human CD1c+dendritic Cells and Enhances Their Toll-like Receptor 7/8-Induced Cytokine Secretion. Front. Immunol. 2018, 9, 744,  DOI: 10.3389/fimmu.2018.00744
      26
      CLEC10A is a specific marker for human CD1c+ dendritic cells and enhances their toll-like receptor 7/8-induced cytokine secretion
      Heger, Lukas; Balk, Silke; Luehr, Jennifer J.; Heidkamp, Gordon F.; Lehmann, Christian H. K.; Hatscher, Lukas; Purbojo, Ariawan; Hartmann, Arndt; Garcia-Martin, Fayna; Nishimura, Shin-Ichiro; Cesnjevar, Robert; Nimmerjahn, Falk; Dudziak, Diana
      Frontiers in Immunology (2018), 9 (), 744/1-744/16CODEN: FIRMCW; ISSN:1664-3224. (Frontiers Media S.A.)
      Dendritic cells (DCs) are major players for the induction of immune responses. Apart from plasmacytoid DCs (pDCs), human DCs can be categorized into two types of conven-tional DCs: CD141+ DCs (cDC1) and CD1c+ DCs (cDC2). Defining uniquely expressed surface markers on human immune cells is not only important for the identification of DC subpopulations but also a prerequisite for harnessing the DC subset-specific potential in immunomodulatory approaches, such as antibody-mediated antigen targeting. Although others identified CLEC9A as a specific endocytic receptor for CD141+ DCs, such a recep-tor for CD1c+ DCs has not been discovered, yet. By performing transcriptomic and flow cytometric analyses on human DC subpopulations from different lymphohematopoietic tissues, we identified CLEC10A (CD301, macrophage galactose-type C-type lectin) as a specific marker for human CD1c+ DCs. We further demonstrate that CLEC10A rapidly internalizes into human CD1c+ DCs upon binding of a monoclonal antibody directed against CLEC10A. The binding of a CLEC10A-specific bivalent ligand (the MUC-1 pep-tide glycosylated with N-acetylgalactosamine) is limited to CD1c+ DCs and enhances the cytokine secretion (namely TNFα, IL-8, and IL-10) induced by TLR 7/8 stimulation. Thus, CLEC10A represents not only a candidate to better define CD1c+ DCs-due to its high endocytic potential-CLEC10A also exhibits an interesting candidate receptor for future antigen-targeting approaches.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXit1Gls7vK&md5=2f4e523c6fac15e319656205747b0255
    27. 27
      Napoletano, C.; Rughetti, A.; Agervig Tarp, M. P.; Coleman, J.; Bennett, E. P.; Picco, G.; Sale, P.; Denda-Nagai, K.; Irimura, T.; Mandel, U.; Clausen, H.; Frati, L.; Taylor-Papadimitriou, J.; Burchell, J.; Nuti, M. Tumor-Associated Tn-MUC1 Glycoform Is Internalized through the Macrophage Galactose-Type C-Type Lectin and Delivered to the HLA Class I and II Compartments in Dendritic Cells. Cancer Res. 2007, 67, 8358– 8367,  DOI: 10.1158/0008-5472.can-07-1035
      27
      Tumor-Associated Tn-MUC1 Glycoform Is Internalized through the Macrophage Galactose-Type C-Type Lectin and Delivered to the HLA Class I and II Compartments in Dendritic Cells
      Napoletano, Chiara; Rughetti, Aurelia; Agervig Tarp, Mads P.; Coleman, Julia; Bennett, Eric P.; Picco, Gianfranco; Sale, Patrizio; Denda-Nagai, Kaori; Irimura, Tatsuro; Mandel, Ulla; Clausen, Henrik; Frati, Luigi; Taylor-Papadimitriou, Joyce; Burchell, Joy; Nuti, Marianna
      Cancer Research (2007), 67 (17), 8358-8367CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
      The type of interaction between tumor-assocd. antigens and specialized antigen-presenting cells such as dendritic cells (DCs) is crit. for the type of immunity that will be generated. MUC1, a highly O-glycosylated mucin, is overexpressed and aberrantly glycosylated in several tumor histiotypes. This results in the expression of tumor-assocd. glycoforms and in MUC1 carrying the tumor-specific glycan Tn (GalNAcα1-O-Ser/Thr). Glycopeptides corresponding to three tandem repeats of MUC1, enzymically glycosylated with 9 or 15 mol of GalNAc, were shown to specifically bind and to be internalized by immature monocyte-derived DCs (iDCs). Binding required calcium and the GalNAc residue and was competed out by GalNAc polymer and Tn-MUC1 or Tn-MUC2 glycopeptides. The macrophage galactose-type C-type lectin (MGL) receptor expressed on iDCs was shown to be responsible for the binding. Confocal anal. and ELISA done on subcellular fractions of iDCs showed that the Tn-MUC1 glycopeptides colocalized with HLA class I and II compartments after internalization. Importantly, although Tn-MUC1 recombinant protein was bound and internalized by MGL, the glycoprotein entered the HLA class II compartment, but not the HLA class I pathway. These data indicate that MGL expressed on iDCs is an optimal receptor for the internalization of short GalNAcs carrying immunogens to be delivered into HLA class I and II compartments. Such glycopeptides therefore represent a new way of targeting the HLA class I and II pathways of DCs. These results have possible implications in designing cancer vaccines.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXpvFKisr4%253D&md5=d1d5372f51e7a10194abd3254298342b
    28. 28
      Napoletano, C.; Zizzari, I. G.; Rughetti, A.; Rahimi, H.; Irimura, T.; Clausen, H.; Wandall, H. H.; Belleudi, F.; Bellati, F.; Pierelli, L.; Frati, L.; Nuti, M. Targeting of Macrophage Galactose-Type C-Type Lectin (MGL) Induces DC Signaling and Activation. Eur. J. Immunol. 2012, 42, 936– 945,  DOI: 10.1002/eji.201142086
      28
      Targeting of macrophage galactose-type C-type lectin (MGL) induces DC signaling and activation
      Napoletano, Chiara; Zizzari, Ilaria G.; Rughetti, Aurelia; Rahimi, Hassan; Irimura, Tatsuro; Clausen, Henrik; Wandall, Hans H.; Belleudi, Francesca; Bellati, Filippo; Pierelli, Luca; Frati, Luigi; Nuti, Marianna
      European Journal of Immunology (2012), 42 (4), 936-945CODEN: EJIMAF; ISSN:0014-2980. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Dendritic cells (DCs) sense the microenvironment through several types of receptors recognizing pathogen-assocd. mol. patterns. In particular, C-type lectins, expressed by distinct subsets of DCs, recognize and internalize specific carbohydrate antigen in a Ca2+-dependent manner. Targeting of these receptors is becoming an efficient strategy of delivering antigens in DC-based anticancer immunotherapy. Here we investigated the role of the macrophage galactose type C-lectin receptor (MGL), expressed by immature DCs (iDCs), as a mol. target for α-N-acetylgalactosamine (GalNAc or Tn)-carrying tumor-assocd. antigens to improve DC performance. MGL expressed by ex vivo-generated iDCs from healthy donors was engaged by a 60-mer MUC19Tn-glycopeptide as a Tn-carrying tumor-assocd. antigen, and an anti-MGL antibody, as a specific MGL binder. We demonstrated that MGL engagement induced homotrimers and homodimers, triggering the phosphorylation of extracellular signal-regulated kinase 1,2 (ERK1,2) and nuclear factor-κB activation. Anal. of DC phenotype and function demonstrated that MGL engagement improved DC performance as antigen-presenting cells, promoting the upregulation of maturation markers, a decrease in phagocytosis, an enhancement of motility, and most importantly an increase in antigen-specific CD8+T-cell activation. These results demonstrate that the targeting of MGL receptor on human DCs has an adjuvant effect and that this strategy can be used to design novel anticancer vaccines.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XmtVKktr0%253D&md5=1937d82644d4875e83734ad6623e849f
    29. 29
      Gabba, A.; Bogucka, A.; Luz, J. G.; Diniz, A.; Coelho, H.; Corzana, F.; Cañada, F. J.; Marcelo, F.; Murphy, P. V.; Birrane, G. Crystal Structure of the Carbohydrate Recognition Domain of the Human Macrophage Galactose C-Type Lectin Bound to GalNAc and the Tumor-Associated Tn Antigen. Biochemistry 2021, 60, 1327– 1336,  DOI: 10.1021/acs.biochem.1c00009
      29
      Crystal Structure of the Carbohydrate Recognition Domain of the Human Macrophage Galactose C-Type Lectin Bound to GalNAc and the Tumor-Associated Tn Antigen
      Gabba, Adele; Bogucka, Agnieszka; Luz, John G.; Diniz, Ana; Coelho, Helena; Corzana, Francisco; Canada, Francisco Javier; Marcelo, Filipa; Murphy, Paul V.; Birrane, Gabriel
      Biochemistry (2021), 60 (17), 1327-1336CODEN: BICHAW; ISSN:0006-2960. (American Chemical Society)
      The human macrophage galactose lectin (MGL) is an endocytic type ii transmembrane receptor expressed on immature monocyte-derived dendritic cells and activated macrophages and plays a role in modulating the immune system in response to infections and cancer. MGL contains an extracellular calcium-dependent (C-type) carbohydrate recognition domain (CRD) that specifically binds terminal N-acetylgalactosamine glycan residues such as the Tn and sialyl-Tn antigens found on tumor cells, as well as other N- and O-glycans displayed on certain viruses and parasites. Even though the glycan specificity of MGL is known and several binding glycoproteins were identified, the mol. basis for substrate recognition has remained elusive due to the lack of high-resoln. structures. Here the authors present crystal structures of the MGL CRD at near endosomal pH and in several complexes, which reveal details of the interactions with the natural ligand, GalNAc, the cancer-assocd. Tn-Ser antigen, and a synthetic GalNAc mimetic ligand. Like the asialoglycoprotein receptor, addnl. calcium atoms are present and contribute to stabilization of the MGL CRD fold. The structure provides the mol. basis for preferential binding of N-acetylgalactosamine over galactose and prompted the reevaluation of the binding modes previously proposed in soln. Satn. transfer difference NMR data acquired using the MGL CRD and interpreted using the crystal structure indicate a single binding mode for GalNAc in soln. Models of MGL1 and MGL2, the mouse homologs of MGL, explain how these proteins might recognize LewisX and GalNAc, resp.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXmsVWjt74%253D&md5=bfc979febdd29a4b74685f1439c2274a
    30. 30
      André, S.; O’Sullivan, S.; Koller, C.; Murphy, P. V.; Gabius, H. J. Bi- to Tetravalent Glycoclusters Presenting GlcNAc/GalNAc as Inhibitors: From Plant Agglutinins to Human Macrophage Galactose-Type Lectin (CD301) and Galectins. Org. Biomol. Chem. 2015, 13, 4190– 4203,  DOI: 10.1039/c5ob00048c
      30
      Bi- to tetravalent glycoclusters presenting GlcNAc/GalNAc as inhibitors: from plant agglutinins to human macrophage galactose-type lectin (CD301) and galectins
      Andre, Sabine; O'Sullivan, Shane; Koller, Christiane; Murphy, Paul V.; Gabius, Hans-Joachim
      Organic & Biomolecular Chemistry (2015), 13 (14), 4190-4203CODEN: OBCRAK; ISSN:1477-0520. (Royal Society of Chemistry)
      Emerging insights into the functional spectrum of tissue lectins leads to identification of new targets for the custom-made design of potent inhibitors, providing a challenge for synthetic chem. The affinity and selectivity of a carbohydrate ligand for a lectin may immensely be increased by a no. of approaches, which includes varying geometrical or topol. features. This perspective leads to the design and synthesis of glycoclusters and their testing using assays of physiol. relevance. Herein, hydroquinone, resorcinol, benzene-1,3,5-triol and tetra(4-hydroxyphenyl)ethene have been employed as scaffolds and propargyl derivs. obtained. The triazole-contg. linker to the α/β-O/S-glycosides of GlcNAc/GalNAc presented on these scaffolds was generated by copper-catalyzed azide-alkyne cycloaddn. This strategy was used to give a panel of nine glycoclusters with bi-, tri- and tetravalency. Maintained activity for lectin binding after conjugation was ascertained for both sugars in solid-phase assays with the plant agglutinins WGA (GlcNAc) and DBA (GalNAc). Absence of cross-reactivity excluded any carbohydrate-independent reactivity of the bivalent compds., allowing the authors to proceed to further testing with a biomedically relevant lectin specific for GalNAc. Macrophage galactose(-binding C)-type lectin, involved in immune defense by dendritic cells and in virus uptake, was produced as a sol. protein without/with its α-helical coiled-coil stalk region. Binding to ligands presented on a matrix and on cell surfaces was highly susceptible to the presence of the tetravalent inhibitor derived from the tetraphenylethene-contg. scaffold, and presentation of GalNAc with an α-thioglycosidic linkage proved favorable. Cross-reactivity of this glycocluster to human galectins-3 and -4, which interact with Tn-antigen-presenting mucins, was rather small. Evidently, the valency and spatial display of α-GalNAc residues is a key factor to design potent and selective inhibitors for this lectin.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXjtVemtbw%253D&md5=0d13d8cd20d4c914636876b34ecee15a
    31. 31
      Kaltner, H.; Manning, J. C.; García Caballero, G.; Di Salvo, C.; Gabba, A.; Romero-Hernández, L. L.; Knospe, C.; Wu, D.; Daly, H. C.; O’Shea, D. F.; Gabius, H. J.; Murphy, P. V. Revealing Biomedically Relevant Cell and Lectin Type-Dependent Structure-Activity Profiles for Glycoclusters by Using Tissue Sections as an Assay Platform. RSC Adv. 2018, 8, 28716– 28735,  DOI: 10.1039/c8ra05382k
      31
      Revealing biomedically relevant cell and lectin type-dependent structure-activity profiles for glycoclusters by using tissue sections as an assay platform
      Kaltner, Herbert; Manning, Joachim C.; Garcia Caballero, Gabriel; Di Salvo, Claudia; Gabba, Adele; Romero-Hernandez, Laura L.; Knospe, Clemens; Wu, Dan; Daly, Harrison C.; O'Shea, Donal F.; Gabius, Hans-Joachim; Murphy, Paul V.
      RSC Advances (2018), 8 (50), 28716-28735CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)
      The increasing realization of the involvement of lectin-glycan recognition in (patho)physiol. processes inspires envisioning therapeutic intervention by high-avidity/specificity blocking reagents. Synthetic glycoclusters are proving to have potential for becoming such inhibitors but the commonly used assays have their drawbacks to predict in vivo efficacy. They do not represent the natural complexity of (i) cell types and (ii) spatial and structural complexity of glycoconjugate representation. Moreover, testing lectins in mixts., as present in situ, remains a major challenge, giving direction to this work. Using a toolbox with four lectins and six bi- to tetravalent glycoclusters bearing the cognate sugar in a model study, we here document the efficient and versatile application of tissue sections (from murine jejunum as the model) as a platform for routine and systematic glycocluster testing without commonly encountered limitations. The nature of glycocluster structure, esp. core and valency, and of protein features, i.e. architecture, fine-specificity and valency, are shown to have an influence, as cell types can differ in response profiles. Proceeding from light microscopy to monitoring by fluorescence microscopy enables grading of glycocluster activity on individual lectins tested in mixts. This work provides a robust tool for testing glycoclusters prior to considering in vivo expts.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhsV2itr3M&md5=54f546a60e3039cd0aeb157e0210375c
    32. 32
      Tseng, N. W.; Liu, J.; Ng, J. C. Y.; Lam, J. W. Y.; Sung, H. H. Y.; Williams, I. D.; Tang, B. Z. Deciphering Mechanism of Aggregation-Induced Emission (AIE): Is E-Z Isomerisation Involved in an AIE Process?. Chem. Sci. 2012, 3, 493– 497,  DOI: 10.1039/c1sc00690h
      32
      Deciphering mechanism of aggregation-induced emission (AIE): Is E-Z isomerisation involved in an AIE process?
      Tseng, Nai-Wen; Liu, Jianzhao; Ng, Jason C. Y.; Lam, Jacky W. Y.; Sung, Herman H. Y.; Williams, Ian D.; Tang, Ben Zhong
      Chemical Science (2012), 3 (2), 493-497CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
      In this work, we address a mechanistic issue on AIE process and correct a long-held misconception on stilbene photoluminescence. E-Z isomerization has been generally recognized as the cause of emission quenching in stilbene solns. A natural question arisen from this common belief is whether suppression of E-Z isomerization by aggregate formation in a stilbenic fluorogen system is responsible for its AIE phenomenon. Monitoring of the structural change of a stilbene deriv. named 1,2-diphenyl-1,2-di(p-tolyl)ethene by NMR during UV irradn. reveals that the E-Z isomerization is not involved in its AIE process under the normal photoluminescence spectral measurement conditions.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XkslaqsQ%253D%253D&md5=e978caacf20d1ce780445f43518662cc
    33. 33
      Donnier-Maréchal, M.; Abdullayev, S.; Bauduin, M.; Pascal, Y.; Fu, M.-Q.; He, X.-P.; Gillon, E.; Imberty, A.; Kipnis, E.; Dessein, R.; Vidal, S. Tetraphenylethylene-Based Glycoclusters with Aggregation-Induced Emission (AIE) Properties as High-Affinity Ligands of Bacterial Lectins. Org. Biomol. Chem. 2018, 16, 8804– 8809,  DOI: 10.1039/c8ob02035c
      33
      Tetraphenylethylene-based glycoclusters with aggregation-induced emission (AIE) properties as high-affinity ligands of bacterial lectins
      Donnier-Marechal, Marion; Abdullayev, Shuay; Bauduin, Marvin; Pascal, Yoann; Fu, Meng-Qi; He, Xiao-Peng; Gillon, Emilie; Imberty, Anne; Kipnis, Eric; Dessein, Rodrigue; Vidal, Sebastien
      Organic & Biomolecular Chemistry (2018), 16 (45), 8804-8809CODEN: OBCRAK; ISSN:1477-0520. (Royal Society of Chemistry)
      Tetraphenylethylene (TPE) is fluorescent through aggregation induced emission (AIE) in water. Herein, TPE was used as the core of glycoclusters that target the bacterial lectins LecA and LecB of Pseudomonas aeruginosa. Synthesis of these TPE-based glycoclusters was accomplished by using azide-alkyne "click" chem. The AIE properties of the resulting glycoclusters could be readily verified, but imaging could not be pursued due to the overlap of the fluorescence signals from cells and bacteria. Nonetheless, the glycoclusters displayed nanomolar affinities toward LecA and LecB. Further evaluation in a cell-based anti-adhesive assay highlighted a limited decrease in adhesion (20%) for the fucosylated glycocluster. This confirmed that these TPE-based glycoclusters are indeed LecA and LecB high-affinity ligands. Nevertheless, the hypotheses involving their application in imaging or anti-adhesive therapy could not be verified.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhvFyntLzJ&md5=df9ac1602341695242757dbd943ff469
    34. 34
      Jégouzo, S. A.; Quintero-Martínez, A.; Ouyang, X.; Dos Santos, Á.; Taylor, M. E.; Drickamer, K. Organization of the Extracellular Portion of the Macrophage Galactose Receptor: A Trimeric Cluster of Simple Binding Sites for N-Acetylgalactosamine. Glycobiology 2013, 23, 853– 864,  DOI: 10.1093/glycob/cwt022
      34
      Organization of the extracellular portion of the macrophage galactose receptor: A trimeric cluster of simple binding sites for N-acetylgalactosamine
      Jegouzo, Sabine A. F.; Quintero-Martinez, Adrian; Ouyang, Xiangyu; dos Santos, Alia; Taylor, Maureen E.; Drickamer, Kurt
      Glycobiology (2013), 23 (7), 853-864CODEN: GLYCE3; ISSN:0959-6658. (Oxford University Press)
      The properties of the human macrophage galactose receptor have been investigated. Specificity for N-acetylgalactosamine (GalNAc) residues with exposed 3- and 4-hydroxyl groups explains virtually all of the results obtained from a recently expanded array of synthetic glycans and is consistent with a model for the structure of the binding site. This simple interaction is sufficient to explain the ability of the receptor to bind to tumor-cell glycans bearing Tn and sialyl-Tn antigens, but not to more elaborate O-linked glycans that predominate on normal cells. This specificity also allows for binding of parasite glycans and screening of an array of bacterial outer membrane oligosaccharides confirms that the receptor binds to a subset of these structures with appropriately exposed GalNAc residues. A key feature of the receptor is the clustering of binding sites in the extracellular portion of the protein, which retains the trimeric structure obsd. in the cell membrane. Chem. crosslinking, gel filtration, CD anal. and differential scanning calorimetry demonstrate that this trimeric structure of the receptor is stabilized by an α-helical coiled coil that extends from the surface of the membrane to the globular carbohydrate-recognition domains. The helical neck domains form independent trimerization domains. Taken together, these results indicate that the macrophage galactose receptor shares many of the features of serum mannose-binding protein, in which clusters of monosaccharide-binding sites serve as detectors for a simple epitope that is not common on endogenous cell surface glycans but that is abundant on the surfaces of tumor cells and certain pathogens.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXptFerurk%253D&md5=dac97ed7fb3dbee78fffe466d1314cfe
    35. 35
      Hu, X. M.; Chen, Q.; Wang, J. X.; Cheng, Q. Y.; Yan, C. G.; Cao, J.; He, Y. J.; Han, B. H. Tetraphenylethylene-Based Glycoconjugate as a Fluorescence “Turn-on” Sensor for Cholera Toxin. Chem.─Asian J. 2011, 6, 2376– 2381,  DOI: 10.1002/asia.201100141
      35
      Tetraphenylethylene-based Glycoconjugate as a Fluorescence "Turn-On" Sensor for Cholera Toxin
      Hu, Xin-Ming; Chen, Qi; Wang, Jin-Xiang; Cheng, Qian-Yi; Yan, Chao-Guo; Cao, Jie; He, Yu-Jian; Han, Bao-Hang
      Chemistry - An Asian Journal (2011), 6 (9), 2376-2381CODEN: CAAJBI; ISSN:1861-4728. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Tetraphenylethylene (TPE)-based glycoconjugates were easily synthesized by copper(I)-catalyzed "click reactions" between propargyl-attached TPE and azido-functionalized sugars. The TPE compd. bearing lactosyl moieties (Lac-TPE) was found to be a fluorescence "turn-on" sensor for cholera toxin by virtue of aggregation-induced emission characteristics of the TPE motif owing to the specific interaction of lactose with the cholera toxin B subunit, while a cellobiose-functionalized TPE deriv. did not show any response to the toxin. Therefore, Lac-TPE shows promising applications in the detection of cholera toxin, as well as in the investigation of carbohydrate-protein interaction.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXoslCrsrY%253D&md5=af3f2d2fede95ef75314655ded79cb5f
    36. 36
      Price, M. R.; Rye, P. D.; Petrakou, E.; Murray, A.; Brady, K.; Imai, S.; Haga, S.; Kiyozuka, Y.; Schol, D.; Meulenbroek, M. F. A.; Snijdewint, F. G. M.; Von Mensdorff-Pouilly, S.; Verstraeten, R. A.; Nil, K.; Blockzjil, A.; Nil, N.; Nilsson, O.; Nil, R.; Suresh, M. R.; Nil, K.; Fortier, S.; Nil, B.; Berg, A.; Longenecker, M. B.; Nil, H.; Boer, M.; Nil, K.; McKenzie, I. F. C.; Nil, G.; Simeoni, L. A.; Ter-Grigoryan, A. G.; Belyanchikov, I. M.; Bovin, N. V.; Cao, Y.; Karsten, U.; Dai, J.; Allard, W. J.; Davis, G.; Yeung, K. K.; Hanisch, F. G.; Lloyd, K. O.; Kudryashov, V.; Sikut, R.; Sikut, A.; Zhang, K.; Baeckström, D.; Hansson, G. C.; Reis, C. A.; Hassan, H.; Bennett, E. P.; Claussen, H.; Norum, L.; Varaas, T.; Kierulf, B.; Nustad, K.; Ciborowski, P.; Konitzki, W. M.; Magarian-Blander, J.; Finn, O. J.; Hilgers, J. Summary Report on the ISOBM TD-4 Workshop: Analysis of 56 Monoclonal Antibodies against the MUC1 Mucin. Tumor Biol. 1998, 19, 1– 20,  DOI: 10.1159/000056500
      36
      Summary report on the ISOBM TD-4 Workshop: analysis of 56 monoclonal antibodies against the MUC1 mucin. San Diego, Calif., November 17-23, 1996
      Price M R; Rye P D; Petrakou E; Murray A; Brady K; Imai S; Haga S; Kiyozuka Y; Schol D; Meulenbroek M F; Snijdewint F G; von Mensdorff-Pouilly S; Verstraeten R A; Kenemans P; Blockzjil A; Nilsson K; Nilsson O; Reddish M; Suresh M R; Koganty R R; Fortier S; Baronic L; Berg A; Longenecker M B; Hilgers J; et al
      Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine (1998), 19 Suppl 1 (), 1-20 ISSN:1010-4283.
      Sixteen research groups participated in the ISOBM TD-4 Workshop in which the reactivity and specificity of 56 monoclonal antibodies against the MUC1 mucin was investigated using a diverse panel of target antigens and MUC1 mucin-related synthetic peptides and glycopeptides. The majority of antibodies (34/56) defined epitopes located within the 20-amino acid tandem repeat sequence of the MUC1 mucin protein core. Of the remaining 22 antibodies, there was evidence for the involvement of carbohydrate residues in the epitopes for 16 antibodies. There was no obvious relationship between the type of immunogen and the specificity of each antibody. Synthetic peptides and glycopeptides were analyzed for their reactivity with each antibody either by assay of direct binding (e.g. by ELISA or BiaCore) or by determining the capacity of synthetic ligands to inhibit antibody binding interactions. There was good concordance between the research groups in identifying antibodies reactive with peptide epitopes within the MUC1 protein core. Epitope mapping tests were performed using the Pepscan analysis for antibody reactivity against overlapping synthetic peptides, and results were largely consistent between research groups. The dominant feature of epitopes within the MUC1 protein core was the presence, in full or part, of the hydrophilic sequence of PDTRAPAP. Carbohydrate epitopes were less easily characterized and the most useful reagents in this respect were defined oligosaccharides, rather than purified mucin preparations enriched in particular carbohydrate moieties. It was evident that carbohydrate residues were involved in many epitopes, by regulating epitope accessibility or masking determinants, or by stabilizing preferred conformations of peptide epitopes within the MUC1 protein core. Overall, the studies, highlight concordance between groups rather than exposing inconsistencies which gives added confidence to the results of analyses of the specificity of antimucin monoclonal antibodies.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADyaK1c%252FosVSjsA%253D%253D&md5=18abcd625acb7d3e72598493a1abc31b
    37. 37
      Pett, C.; Cai, H.; Liu, J.; Palitzsch, B.; Schorlemer, M.; Hartmann, S.; Stergiou, N.; Lu, M.; Kunz, H.; Schmitt, E.; Westerlind, U. Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. Chem.─Eur. J. 2017, 23, 3875– 3884,  DOI: 10.1002/chem.201603921
      37
      Microarray analysis of antibodies induced with synthetic antitumor vaccines: Specificity against diverse mucin core structures
      Pett, Christian; Cai, Hui; Liu, Jia; Palitzsch, Bjoern; Schorlemer, Manuel; Hartmann, Sebastian; Stergiou, Natascha; Lu, Mengji; Kunz, Horst; Schmitt, Edgar; Westerlind, Ulrika
      Chemistry - A European Journal (2017), 23 (16), 3875-3884CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Glycoprotein research is pivotal for vaccine development and biomarker discovery. Many successful methodologies for reliably increasing the antigenicity toward tumor-assocd. glycopeptide structures have been reported. Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunol. memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunol. For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-assocd. glycans (TN, STN, and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable d. at the five different sites of the MUC1 tandem repeat. This is one of the most extensive glycopeptide libraries ever made through total synthesis. On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amts. of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-assocd. antigen structures. The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. It is obsd. that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. All antibody sera show high reactivity to the tumor-assocd. saccharide structures on MUC1. Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhsVaqs7c%253D&md5=6af0f627abe8d7c19adef5f7c7cea23a
    38. 38
      Wandall, H. H.; Blixt, O.; Tarp, M. A.; Pedersen, J. W.; Bennett, E. P.; Mandel, U.; Ragupathi, G.; Livingston, P. O.; Hollingsworth, M. A.; Taylor-Papadimitriou, J.; Burchell, J.; Clausen, H. Cancer Biomarkers Defined by Autoantibody Signatures to Aberrant O-Glycopeptide Epitopes. Cancer Res. 2010, 70, 1306– 1313,  DOI: 10.1158/0008-5472.can-09-2893
      38
      Cancer Biomarkers Defined by Autoantibody Signatures to Aberrant O-Glycopeptide Epitopes
      Wandall, Hans H.; Blixt, Ola; Tarp, Mads A.; Pedersen, Johannes W.; Bennett, Eric P.; Mandel, Ulla; Ragupathi, Govind; Livingston, Phil O.; Hollingsworth, Michael A.; Taylor-Papadimitriou, Joyce; Burchell, Joy; Clausen, Henrik
      Cancer Research (2010), 70 (4), 1306-1313CODEN: CNREA8; ISSN:0008-5472. (American Association for Cancer Research)
      Autoantibodies to cancer antigens hold promise as biomarkers for early detection of cancer. Proteins that are aberrantly processed in cancer cells are likely to present autoantibody targets. The extracellular mucin MUC1 is overexpressed and aberrantly glycosylated in many cancers; thus, the authors evaluated whether autoantibodies generated to aberrant O-glycoforms of MUC1 might serve as sensitive diagnostic biomarkers for cancer. Using an antibody-based glycoprofiling ELISA assay, the authors documented that aberrant truncated glycoforms were not detected in sera of cancer patients. An O-glycopeptide microarray was developed that detected IgG antibodies to aberrant O-glycopeptide epitopes in patients vaccinated with a keyhole limpet hemocyanin-conjugated truncated MUC1 peptide. The authors detected cancer-assocd. IgG autoantibodies in sera from breast, ovarian, and prostate cancer patients against different aberrent O-glycopeptide epitopes derived from MUC1. These autoantibodies represent a previously unaddressed source of sensitive biomarkers for early detection of cancer. The methods the authors have developed for chemoenzymic synthesis of O-glycopeptides on microarrays may allow for broader mining of the entire cancer O-glycopeptidome. Cancer Res; 70(4); 1306-13.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhvFaksrc%253D&md5=f21b0b73d10e93808720c84f4f4c8711
    39. 39
      Borgert, A.; Heimburg-Molinaro, J.; Song, X.; Lasanajak, Y.; Ju, T.; Liu, M.; Thompson, P.; Ragupathi, G.; Barany, G.; Smith, D. F.; Cummings, R. D.; Live, D. Deciphering Structural Elements of Mucin Glycoprotein Recognition. ACS Chem. Biol. 2012, 7, 1031– 1039,  DOI: 10.1021/cb300076s
      39
      Deciphering Structural Elements of Mucin Glycoprotein Recognition
      Borgert, Andrew; Heimburg-Molinaro, Jamie; Song, Xuezheng; Lasanajak, Yi; Ju, Tongzhong; Liu, Mian; Thompson, Pamela; Ragupathi, Govind; Barany, George; Smith, David F.; Cummings, Richard D.; Live, David
      ACS Chemical Biology (2012), 7 (6), 1031-1039CODEN: ACBCCT; ISSN:1554-8929. (American Chemical Society)
      Mucin glycoproteins present a complex structural landscape arising from the multiplicity of glycosylation patterns afforded by their numerous serine and threonine glycosylation sites, often in clusters, and with variations in resp. glycans. To explore the structural complexities in such glycoconjugates, the authors used NMR to systematically analyze the conformational effects of glycosylation d. within a cluster of sites. This allows correlation with mol. recognition through anal. of interactions between these and other glycopeptides, with antibodies, lectins, and sera, using a glycopeptide microarray. Selective antibody interactions with discrete conformational elements, reflecting aspects of the peptide and disposition of GalNAc residues, are obsd. The authors' results help bridge the gap between conformational properties and mol. recognition of these mols., with implications for their physiol. roles. Features of the native mucin motifs impact their relative immunogenicity and are accurately encoded in the antibody binding site, with the conformational integrity being preserved in isolated glycopeptides, as reflected in the antibody binding profile to array components.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XksVeksrw%253D&md5=c37329341e9dd4ef7681a22906532ea1
    40. 40
      Coltart, D. M.; Royyuru, A. K.; Williams, L. J.; Glunz, P. W.; Sames, D.; Kuduk, S. D.; Schwarz, J. B.; Chen, X. T.; Danishefsky, S. J.; Live, D. H. Principles of Mucin Architecture: Structural Studies on Synthetic Glycopeptides Bearing Clustered Mono-Di-Tri-and Hexasaccharide Glycodomains. J. Am. Chem. Soc. 2002, 124, 9833– 9844,  DOI: 10.1021/ja020208f
      40
      Principles of Mucin Architecture: Structural Studies on Synthetic Glycopeptides Bearing Clustered Mono-, Di-, Tri-, and Hexasaccharide Glycodomains
      Coltart, Don M.; Royyuru, Ajay K.; Williams, Lawrence J.; Glunz, Peter W.; Sames, Dalibor; Kuduk, Scott D.; Schwarz, Jacob B.; Chen, Xiao-Tao; Danishefsky, Samuel J.; Live, David H.
      Journal of the American Chemical Society (2002), 124 (33), 9833-9844CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      The structural characteristics of a mucin glycopeptide motif derived from the N-terminal fragment STTAV of the cell surface glycoprotein CD43 have been investigated by NMR. In this study, a series of mols. prepd. by total synthesis were examd., consisting of the peptide itself, three glycopeptides having clustered sites of α-O-glycosylation on the serine and threonine side chains with the Tn, TF, and STF carbohydrate antigens, resp., and one with the β-O-linked TF antigen. Addnl., a glycopeptide having the sequence SSSAVAV, triglycosylated with the Ley epitope, was investigated. NMR data for the tri-STF-STTAV glycopeptide were used to solve the structure of this construct through restrained mol. dynamics calcns. The calcns. revealed a defined conformation for the glycopeptide core rooted in the interaction of the peptide and the first N-acetylgalactosamine residue. The similarity of the NMR data for each of the α-O-linked glycopeptides demonstrates that this structure persists for each construct and that the mode of attachment of the first sugar and the peptide is paramount in establishing the organization of the core. The core provides a common framework on which a variety of glycans may be displayed. Remarkably, while there is a profound organizational effect on the peptide backbone with the α-linked glycans, attachment via a β-linkage has little apparent consequence.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XlsFygtL8%253D&md5=db84e77f38b2cba8d3645af74b80257f
    41. 41
      Apostolopoulos, V.; Yuriev, E.; Ramsland, P. A.; Halton, J.; Osinski, C.; Li, W.; Plebanski, M.; Paulsen, H.; McKenzie, I. F. C. A Glycopeptide in Complex with MHC Class I Uses the GalNAc Residue as an Anchor. Proc. Natl. Acad. Sci. U.S.A. 2003, 100, 15029– 15034,  DOI: 10.1073/pnas.2432220100
      41
      A glycopeptide in complex with MHC class I uses the GalNAc residue as an anchor
      Apostolopoulos, Vasso; Yuriev, Elizabeth; Ramsland, Paul A.; Halton, Jodie; Osinski, Carla; Li, Wenjun; Plebanski, Magdalena; Paulsen, Hans; McKenzie, Ian F. C.
      Proceedings of the National Academy of Sciences of the United States of America (2003), 100 (25), 15029-15034CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
      Peptides bind MHC class I mols. by anchoring hydrophobic side chains into pockets in the peptide binding groove. Here, we report an immunogenic (in vitro and in vivo) MUC1 glycopeptide (MUC1-8-5GalNAc) bound to H-2Kb, fully crossreactive with the nonglycosylated variant. Mol. modeling showed that the central P5-Thr-GalNAc residue points into the C pocket and forms van der Waals and hydrogen bond interactions with the MHC class I. As predicted, GalNAc, a modified peptide carrying an addnl. anchor in the central C anchor pocket, increased the affinity by ≈100-fold compared with the native low-affinity peptide (MUC1-8). The findings demonstrate that glycopeptides assocd. with MHC class I mols. can use GalNAc to anchor the peptide in the groove and enable high-affinity binding.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXpvFaqtLY%253D&md5=22feb349b128c699a317d5d103262948
    42. 42
      Avci, F. Y.; Li, X.; Tsuji, M.; Kasper, D. L. A Mechanism for Glycoconjugate Vaccine Activation of the Adaptive Immune System and Its Implications for Vaccine Design. Nat. Med. 2011, 17, 1602– 1609,  DOI: 10.1038/nm.2535
      42
      A mechanism for glycoconjugate vaccine activation of the adaptive immune system and its implications for vaccine design
      Avci, Fikri Y.; Li, Xiangming; Tsuji, Moriya; Kasper, Dennis L.
      Nature Medicine (New York, NY, United States) (2011), 17 (12), 1602-1609CODEN: NAMEFI; ISSN:1078-8956. (Nature Publishing Group)
      Glycoconjugate vaccines have provided enormous health benefits globally, but they have been less successful in some populations at high risk for developing disease. To identify new approaches to enhancing glycoconjugate effectiveness, we investigated mol. and cellular mechanisms governing the immune response to a prototypical glycoconjugate vaccine. We found that in antigen-presenting cells a carbohydrate epitope is generated upon endolysosomal processing of group B streptococcal type III polysaccharide coupled to a carrier protein. In conjunction with a carrier protein-derived peptide, this carbohydrate epitope binds major histocompatibility class II (MHCII) and stimulates carbohydrate-specific CD4+ T cell clones to produce interleukins 2 and 4-cytokines essential for providing T cell help to antibody-producing B cells. An archetypical glycoconjugate vaccine that we constructed to maximize the presentation of carbohydrate-specific T cell epitopes is 50-100 times more potent and substantially more protective in a neonatal mouse model of group B Streptococcus infection than a vaccine constructed by methods currently used by the vaccine industry. Our discovery of how glycoconjugates are processed resulting in presentation of carbohydrate epitopes that stimulate CD4+ T cells has key implications for glycoconjugate vaccine design that could result in greatly enhanced vaccine efficacy.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsV2gt7%252FL&md5=c02d36037865b38be13179ca1eb84fce
    43. 43
      Vinuesa, C. G.; Linterman, M. A.; Yu, D.; Maclennan, I. C. M. Follicular Helper T Cells. Annu. Rev. Immunol. 2016, 34, 335– 368,  DOI: 10.1146/annurev-immunol-041015-055605
      43
      Follicular Helper T Cells
      Vinuesa, Carola G.; Linterman, Michelle A.; Yu, Di; MacLennan, Ian C. M.
      Annual Review of Immunology (2016), 34 (), 335-368CODEN: ARIMDU; ISSN:0732-0582. (Annual Reviews)
      Although T cell help for B cells was described several decades ago, it was the identification of CXCR5 expression by B follicular helper T (Tfh) cells and the subsequent discovery of their dependence on BCL6 that led to the recognition of Tfh cells as an independent helper subset and accelerated the pace of discovery. More than 20 transcription factors, together with RNA-binding proteins and microRNAs, control the expression of chemotactic receptors and mols. important for the function and homeostasis of Tfh cells. Tfh cells prime B cells to initiate extrafollicular and germinal center antibody responses and are crucial for affinity maturation and maintenance of humoral memory. In addn. to the roles that Tfh cells have in antimicrobial defense, in cancer, and as HIV reservoirs, regulation of these cells is crit. to prevent autoimmunity. The realization that follicular T cells are heterogeneous, comprising helper and regulatory subsets, has raised questions regarding a possible division of labor in germinal center B cell selection and elimination.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XivFKlsrw%253D&md5=4c8dc98065906b9115c3876d236f16be
    44. 44
      Xie, Y.; Chen, Y.; Ahmed, K. A.; Li, W.; Ahmed, S.; Sami, A.; Chibbar, R.; Tang, X.; Tao, M.; Xu, J.; Xiang, J. Potent CD4+ T-Cell Epitope P30 Enhances HER2/Neu-Engineered Dendritic Cell-Induced Immunity against Tg1-1 Breast Cancer in Transgenic FVBneuN Mice by Enhanced CD4+ T-Cell-Stimulated CTL Responses. Cancer Gene Ther. 2013, 20, 590– 598,  DOI: 10.1038/cgt.2013.60
      44
      Potent CD4+ T-cell epitope P30 enhances HER2/neu-engineered dendritic cell-induced immunity against Tg1-1 breast cancer in transgenic FVBneuN mice by enhanced CD4+ T-cell-stimulated CTL responses
      Xie, Y.; Chen, Y.; Ahmed, K. A.; Li, W.; Ahmed, S.; Sami, A.; Chibbar, R.; Tang, X.; Tao, M.; Xu, J.; Xiang, J.
      Cancer Gene Therapy (2013), 20 (10), 590-598CODEN: CGTHEG; ISSN:0929-1903. (Nature Publishing Group)
      One of the major obstacles in human epidermal growth factor receptor (HER)-2/neu-specific trastuzumab immunotherapy of HER2/neu-pos. breast cancer is the development of trastuzumab resistance, warranting the search for other therapeutic strategies. Although dendritic cell (DC) vaccines have been extensively applied in clin. trials for cancer treatment, the vaccination efficacy is still limited, mostly because DC vaccines are not sufficient to break tumor-assocd. antigen-specific self-immune tolerance in cancer patients. P30 (FNNFTVSFWLRVPKVSASHLE) derived from tetanus toxin is a universally potent CD4+ T helper epitope capable of enhancing CD8+ cytotoxic T-lymphocyte (CTL) responses. In this study, the authors constructed two recombinant adenoviral vectors (AdVs), AdVOVA-P30 and AdVHER2/neu-P30, expressing ovalbumin (OVA)-P30 and HER2/neu-P30. To enhance DC vaccine efficacy, the authors transfected mouse bone marrow (BM)-derived DCs with AdVOVA-P30 and AdVHER2/neu-P30 to generate engineered DCOVA-P30 and DCHER2/neu-P30 vaccines, resp. The authors, then, compared CD4+ and CD8+ T-cell responses and antitumor immunity derived from DCOVA-P30 and DCHER2/neu-P30 vaccination in wild-type C57BL/6 and transgenic FVBneuN mice, resp. The authors demonstrate that engineered DCOVA-P30 vaccine stimulates more efficient CD4+ and CD8+ T-cell responses than DCOVA in C57BL/6 mice. Interestingly, the increased DCOVA-P30-induced CTL responses are mainly contributed by enhanced CD4+ T-cell-stimulated CTL proliferation. The authors show that DCOVA-P30 vaccine also stimulates more efficient therapeutic immunity against OVA-expressing BL6-10OVA melanoma than DCOVA in C57BL/6 mice. In addn., the authors demonstrate that DCHER2/neu-P30 vaccine stimulates more efficient CD4+ and CD8+ T-cell responses and protective immunity against HER2/neu-expressing Tg1-1 breast cancer than DCHER2/neu in transgenic FVBneuN mice with HER2/neu-specific self-immune tolerance. Therefore, the engineered DCHER2/neu-P30 vaccine may provide a new immunotherapy alternative for women with HER2/neu+ breast cancer, esp. for trastuzumab-resistant HER2/neu+ breast cancer patients.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVynurvK&md5=41b0969396d251e48b29472f55e361df
    45. 45
      Pett, C.; Schorlemer, M.; Westerlind, U. A Unified Strategy for the Synthesis of Mucin Cores 1-4 Saccharides and the Assembled Multivalent Glycopeptides. Chem.─Eur. J. 2013, 19, 17001– 17010,  DOI: 10.1002/chem.201302921
      45
      A unified strategy for the synthesis of mucin cores 1-4 saccharides and the assembled multivalent glycopeptides
      Pett, Christian; Schorlemer, Manuel; Westerlind, Ulrika
      Chemistry - A European Journal (2013), 19 (50), 17001-17010CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
      By displaying different O-glycans in a multivalent mode, mucin and mucin-like glycoproteins are involved in a plethora of protein binding events. The understanding of the roles of the glycans and the identification of potential glycan binding proteins are major challenges. To enable future binding studies of mucin glycan and glycopeptide probes, a method that gives flexible and efficient access to all common mucin core-glycosylated amino acids was developed. Based on a convergent synthesis strategy starting from a shared early stage intermediate by differentiation in the glycoside acceptor reactivity, a common disaccharide building block allows for the creation of extended glycosylated amino acids carrying the mucin type-2 cores 1-4 saccharides. Formation of a phenyl-sulfenyl-N-Troc (Troc = trichloroethoxycarbonyl) byproduct during N-iodosuccinimide-promoted thioglycoside couplings was further characterized and a new methodol. for the removal of the Troc group is described. The obtained glycosylated 9-fluorenylmethoxycarbonyl (Fmoc)-protected amino acid building blocks are incorporated into peptides for multivalent glycan display.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhslaqsL3E&md5=76a4e68e8e97ba1fc418a5a6e3968d89
    46. 46
      Pett, C.; Westerlind, U. A Convergent Strategy for the Synthesis of Type-1 Elongated Mucin Cores 1-3 and the Corresponding Glycopeptides. Chem.─Eur. J. 2014, 20, 7287– 7299,  DOI: 10.1002/chem.201400162
      46
      A Convergent Strategy for the Synthesis of Type-1 Elongated Mucin Cores 1-3 and the Corresponding Glycopeptides
      Pett, Christian; Westerlind, Ulrika
      Chemistry - A European Journal (2014), 20 (24), 7287-7299CODEN: CEUJED; ISSN:0947-6539. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Mucins are a class of highly O-glycosylated proteins found on the surface of cells in epithelial tissues. O-Glycosylation is crucial for the functionality of mucins and changes therein can have severe consequences for an organism. With that in mind, the elucidation of interactions of carbohydrate binding proteins with mucins, whether in morbidly altered or unaltered conditions, continue to shed light on mechanisms involved in diseases like chronic inflammations and cancer. Despite the known importance of type-1 and type-2 elongated mucin cores 1-4 in glycobiol., the corresponding type-1 structures are much less well studied. Here, the first chem. synthesis of extended mucin type-1 O-glycan core 1-3 amino acid structures based on a convergent approach is presented. By utilizing differentiation in acceptor reactivity, shared early stage Tn- and T-acceptor intermediates were elongated with a common type-1 [β-D-Gal-1,3-β-D-GlcNAc] disaccharide, which allows for straightforward prepn. of diverse glycosylated amino acids carrying the type-1 mucin core 1-3 saccharides. The obtained glycosylated 9-fluorenylmethoxycarbonyl (Fmoc)-protected amino acid building blocks were employed in synthesis of type-1 mucin glycopeptides, which are useful in biol. applications.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXot1Sitbc%253D&md5=7ae3edc6aada767e8621be31268fd418
    47. 47
      Li, R.-J. E.; Hogervorst, T. P.; Achilli, S.; Bruijns, S. C. M.; Spiekstra, S.; Vivès, C.; Thépaut, M.; Filippov, D. V.; van der Marel, G. A.; van Vliet, S. J.; Fieschi, F.; Codée, J. D. C.; van Kooyk, Y. Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens. Front. Cell Dev. Biol. 2020, 8, 556,  DOI: 10.3389/fcell.2020.00556
      47
      Targeting of the C-Type Lectin Receptor Langerin Using Bifunctional Mannosylated Antigens
      Li Rui-Jun Eveline; Bruijns Sven C M; Spiekstra Sander; van Vliet Sandra J; van Kooyk Yvette; Hogervorst Tim P; Filippov Dmitri V; van der Marel Gijs A; Codee Jeroen D C; Achilli Silvia; Vives Corinne; Thepaut Michel; Fieschi Franck
      Frontiers in cell and developmental biology (2020), 8 (), 556 ISSN:2296-634X.
      Langerhans cells (LCs) are antigen-presenting cells that reside in the skin. They uniquely express high levels of the C-type lectin receptor Langerin (CD207), which is an attractive target for antigen delivery in immunotherapeutic vaccination strategies against cancer. We here assess a library of 20 synthetic, well-defined mannoside clusters, built up from one, two, and three of six monomannosides, dimannosides, or trimannosides, appended to an oligopeptide backbone, for binding with Langerin using surface plasmon resonance and flow cytometric quantification. It is found that Langerin binding affinity increases with increasing number of mannosides. Hexavalent presentation of the mannosides resulted in binding affinities ranging from 3 to 12 μM. Trivalent presentation of the dimannosides and trimannosides led to Langerin affinity in the same range. The model melanoma gp100 antigenic peptide was subsequently equipped with a hexavalent cluster of the dimannosides and trimannosides as targeting moieties. Surprisingly, although the bifunctional conjugates were taken up in LCs in a Langerin-dependent manner, limited antigen presentation to cytotoxic T cells was observed. These results indicate that targeting glycan moieties on immunotherapeutic vaccines should not only be validated for target binding, but also on the continued effects on biology, such as antigen presentation to both CD8(+) and CD4(+) T cells.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB38fitVaqtA%253D%253D&md5=d0aefd6bb1ad3f4b602a33cb65d76342
    48. 48
      Li, R.-J. E.; Hogervorst, T. P.; Achilli, S.; Bruijns, S. C.; Arnoldus, T.; Vivès, C.; Wong, C. C.; Thépaut, M.; Meeuwenoord, N. J.; van den Elst, H.; Overkleeft, H. S.; van der Marel, G. A.; Filippov, D. V.; van Vliet, S. J.; Fieschi, F.; Codée, J. D. C.; van Kooyk, Y. Systematic Dual Targeting of Dendritic Cell C-Type Lectin Receptor DC-SIGN and TLR7 Using a Trifunctional Mannosylated Antigen. Front. Chem. 2019, 7, 650,  DOI: 10.3389/fchem.2019.00650
      48
      Systematic dual targeting of dendritic cell C-type lectin receptor DC-SIGN and TLR7 using a trifunctional mannosylated antigen
      Li, Rui-Jun Eveline; Hogervorst, Tim P.; Achilli, Silvia; Bruijns, Sven C.; Arnoldus, Tim; Vives, Corinne; Wong, Chung C.; Thepaut, Michel; Meeuwenoord, Nico J.; van den Elst, Hans; Overkleeft, Herman S.; van der Marel, Gijs A.; Filippov, Dmitri V.; van Vliet, Sandra J.; Fieschi, Franck; Codee, Jeroen D. C.; van Kooyk, Yvette
      Frontiers in Chemistry (Lausanne, Switzerland) (2019), 7 (), 650CODEN: FCLSAA; ISSN:2296-2646. (Frontiers Media S.A.)
      Dendritic cells are important initiators of adaptive immunity, and they possess a multitude of Pattern Recognition Receptors to generate an adequate T cell mediated immunity against invading pathogens. PRR ligands are frequently conjugated to tumor-assocd. antigens in a vaccination strategy to enhance the immune response toward such antigens. One of these PPRs, DC-SIGN, a member of the C-type lectin receptor family, has been extensively targeted with Lewis structures and mannose glycans, often presented in multivalent fashion. Hexavalent presentation of the clusters showed the highest binding affinity, with the hexa-α1,2-di-mannoside being the most potent ligand. The four highest binding hexavalent mannoside structures were conjugated to a model melanoma gp100-peptide antigen and further equipped with a Toll-like receptor 7 (TLR7)-agonist as adjuvant for DC maturation, creating a trifunctional vaccine conjugate. Interestingly, DC-SIGN affinity of the mannoside clusters did not directly correlate with antigen presentation enhancing properties and the α1,2-di-mannoside cluster with the highest binding affinity in our library even hampered T cell activation. Overall, this systematic study has demonstrated that multivalent glycan presentation can improve DC-SIGN binding but enhanced binding cannot be directly translated into enhanced antigen presentation and the sole assessment of binding affinity is thus insufficient to det. further functional biol. activity.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXps1Crs7w%253D&md5=1857ff8a25c82901090efae7c11e215a
    49. 49
      Wu, X.; McKay, C.; Pett, C.; Yu, J.; Schorlemer, M.; Ramadan, S.; Lang, S.; Behren, S.; Westerlind, U.; Finn, M. G.; Huang, X. Synthesis and Immunological Evaluation of Disaccharide Bearing MUC-1 Glycopeptide Conjugates with Virus-like Particles. ACS Chem. Biol. 2019, 14, 2176– 2184,  DOI: 10.1021/acschembio.9b00381
      49
      Synthesis and immunological evaluation of disaccharide bearing MUC-1 glycopeptide conjugates with virus-like particles
      Wu, Xuanjun; McKay, Craig; Pett, Christian; Yu, Jin; Schorlemer, Manuel; Ramadan, Sherif; Lang, Shuyao; Behren, Sandra; Westerlind, Ulrika; Finn, M. G.; Huang, Xuefei
      ACS Chemical Biology (2019), 14 (10), 2176-2184CODEN: ACBCCT; ISSN:1554-8929. (American Chemical Society)
      Mucin-1 (MUC1) is a highly attractive antigenic target for anticancer vaccines. Naturally existing MUC1 can contain multiple types of O-linked glycans, including the Thomsen-Friedenreich (Tf) antigen and the Sialyl Thomsen-nouveau (STn) antigen. In order to target these antigens as potential anticancer vaccines, MUC1 glycopeptides SAPDT*RPAP (T* is the glycosylation site) bearing the Tf and the STn antigen, resp., have been synthesized. The bacteriophage Qβ carrier is a powerful carrier for antigen delivery. The conjugates of MUC1-Tf and -STn glycopeptides with Qβ were utilized to immunize immune-tolerant human MUC1 transgenic (MUC1.Tg) mice, which elicited superior levels of anti-MUC1 IgG antibodies with titers reaching over 2 million units. The IgG antibodies recognized a wide range of MUC1 glycopeptides bearing diverse glycans. Antibodies induced by Qβ-MUC1-Tf showed strongest binding, with MUC1-expressing melanoma B16-MUC1 cells, and effectively killed these cells in vitro. Vaccination with Qβ-MUC1-Tf first followed by tumor challenge in a lung metastasis model showed significant redns. of the no. of tumor foci in the lungs of immunized mice as compared to those in control mice. This was the first time that a MUC1-Tf-based vaccine has shown in vivo efficacy in a tumor model. As such, Qβ-MUC1 glycopeptide conjugates have great potential as anticancer vaccines.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhslegtL3E&md5=bdf9d26dd58bd13745dc5219ff444017
    50. 50
      Pirro, M.; Schoof, E.; Van Vliet, S. J.; Rombouts, Y.; Stella, A.; De Ru, A.; Mohammed, Y.; Wuhrer, M.; Van Veelen, P. A.; Hensbergen, P. J. Glycoproteomic Analysis of MGL-Binding Proteins on Acute T-Cell Leukemia Cells. J. Proteome Res. 2019, 18, 1125– 1132,  DOI: 10.1021/acs.jproteome.8b00796
      50
      Glycoproteomic Analysis of MGL-Binding Proteins on Acute T-Cell Leukemia Cells
      Pirro, Martina; Schoof, Esmee; van Vliet, Sandra J.; Rombouts, Yoann; Stella, Alexandre; de Ru, Arnoud; Mohammed, Yassene; Wuhrer, Manfred; van Veelen, Peter A.; Hensbergen, Paul J.
      Journal of Proteome Research (2019), 18 (3), 1125-1132CODEN: JPROBS; ISSN:1535-3893. (American Chemical Society)
      C-type lectins are a diverse group of proteins involved in many human physiol. and pathol. processes. Most C-type lectins are glycan-binding proteins, some of which are pivotal for innate immune responses against pathogens. Other C-type lectins, such as the macrophage galactose-type lectin (MGL), have been shown to induce immunosuppressive responses upon the recognition of aberrant glycosylation on cancer cells. MGL is known to recognize terminal N-acetylgalactosamine (GalNAc), such as the Tn antigen, which is commonly found on malignant cells. Even though this glycan specificity of MGL is well described, there is a lack of understanding of the actual glycoproteins that bind MGL. We present a glycoproteomic workflow for the identification of MGL-binding proteins, which we applied to study MGL ligands on the human Jurkat leukemia cell line. In addn. to the known MGL ligands and Tn antigen-carrying proteins CD43 and CD45 on these cells, we have identified a set of novel cell-surface ligands for MGL. Importantly, for several of these, O-glycosylation has hitherto not been described. Altogether, our data provide new insight into the identification and structure of novel MGL ligands that presumably act as modulatory mols. in cancer immune responses.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXisFyksrjE&md5=b9d3814d8c244b0c9528e4dc6b3370f6
    51. 51
      Seder, R.; Reed, S. G.; O’Hagan, D.; Malyala, P.; D’Oro, U.; Laera, D.; Abrignani, S.; Cerundolo, V.; Steinman, L.; Bertholet, S. Gaps in Knowledge and Prospects for Research of Adjuvanted Vaccines. Vaccine 2015, 33, B40– B43,  DOI: 10.1016/j.vaccine.2015.03.057
      There is no corresponding record for this reference.
    52. 52
      Hanisch, F.-G.; Stadie, T.; Deutzmann, F.; Peter-Katalinic, J. MUC1 Glycoforms in Breast Cancer. Cell Line T47D as a Model for Carcinoma-Associated Alterations of O-Glycosylation. Eur. J. Biochem. 1996, 236, 318– 327,  DOI: 10.1111/j.1432-1033.1996.00318.x
      52
      MUC1 glycoforms in breast cancer. Cell line T47D as a model for carcinoma-associated alterations of O-glycosylation
      Hanisch, Franz-Georg; Stadie, Tanja R. E.; Deutzmann, Frank; Peter-Katalinic, Jasna
      European Journal of Biochemistry (1996), 236 (1), 318-27CODEN: EJBCAI; ISSN:0014-2956. (Springer)
      A highly immunogenic peptide motif within the tandem repeat domain of MUC1 mucin is assumed to be exposed during development of breast cancer due to altered O-glycosylation. To elucidate the structural aspects of these changes, the authors have isolated and analyzed the integrated or secretory MUC1 glycoforms from carcinoma cell lines or solid tumors and from human milk. The buoyant densities measured in CsCl gradients for MUC1 glycoforms from cancer cells revealed heterogeneity of the physicochem. species and a significant redn. of their carbohydrate contents compared to MUC1 from skim milk. Immunoreactivity patterns of MUC1 glycoforms from tumor or T47D cells exhibited a lack of fucosylated Lewis blood-group-related antigens and the appearance of core-type antigen sialyl-(NeuGl)-TF, Galβ1-3(NeuGlα2-6)GalNAc. Structural chem. of MUC1 oligosaccharides demonstrated that the cancer-assocd. glycoforms carry mainly sialylated trisaccharides NeuAcα2-3Galβ1-3GalNAc or NeuAcα2-6(Galβ1-3)GalNAc, exhibit a concomitant decrease in the ratio of GlcNAc/GalNAc, a redn. or disappearance of L-fucose, and a partial substitution of N-acetylneuraminic acid by the N-glycolylated variant. On comparison to the secretory MUC1 in human milk, the glycoforms on human milk fat globule membranes showed apparently identical patterns of O-linked oligosaccharides with a preponderance of neutral polylactosamino-glycans. During serum-free cultivation of T47D cells over 4 wk, the expression of secretory MUC1 glycoforms was inconsistent based on the decreasing contents of sialic acid and on the concomitant increase of immunodetectable TF antigen.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28Xhtlantrs%253D&md5=24687001dc9ac24202033c516bf4ec5a
    53. 53
      Stergiou, N.; Nagel, J.; Pektor, S.; Heimes, A. S.; Jäkel, J.; Brenner, W.; Schmidt, M.; Miederer, M.; Kunz, H.; Roesch, F.; Schmitt, E. Evaluation of a Novel Monoclonal Antibody against Tumor-Associated MUC1 for Diagnosis and Prognosis of Breast Cancer. Int. J. Med. Sci. 2019, 16, 1188– 1198,  DOI: 10.7150/ijms.35452
      53
      Evaluation of a novel monoclonal antibody against tumor-associated MUC1 for diagnosis and prognosis of breast cancer
      Stergiou, Natascha; Nagel, Johannes; Pektor, Stefanie; Heimes, Anne-Sophie; Jaekel, Joerg; Brenner, Walburgis; Schmt, Marcus; Miederer, Matthias; Kunz, Horst; Roesch, Frank; Schmitt, Edgar
      International Journal of Medical Sciences (2019), 16 (9), 1188-1198CODEN: IJMSGZ; ISSN:1449-1907. (Ivyspring International Publisher)
      There is still a great unmet medical need concerning diagnosis and treatment of breast cancer which could be addressed by utilizing specific mol. targets. Tumor-assocd. MUC1 is expressed on over 90 % of all breast cancer entities and differs strongly from its physiol. form on epithelial cells, therefore presenting a unique target for breast cancer diagnosis and antibody-mediated immune therapy. Utilizing an anti-tumor vaccine based on a synthetically prepd. glycopeptide, we generated a monoclonal antibody (mAb) GGSK-1/30, selectively recognizing human tumor-assocd. MUC1. This antibody targets exclusively tumor-assocd. MUC1 in the absence of any binding to MUC1 on healthy epithelial cells thus enabling the generation of breast tumor-specific radiolabeled immune therapeutic tools. Methods: MAb GGSK-1/30 was used for immunohistochem. anal. of human breast cancer tissue. Its desferrioxamine (Df')-conjugate was synthesized and labeled with 89Zr. [89Zr]Zr-Df'-GGSK-1/30 was evaluated as a potential PET tracer. Binding and pharmacokinetic properties of [89Zr]Zr-Df'-GGSK-1/30 were analyzed in vitro using human and murine cell lines that express tumor-assocd. MUC1. Self-generated primary murine breast cancer cells expressing human tumor-assocd. MUC1 were transplanted s.c. in wild type and human MUC1-transgenic mice. The pharmacol. of [89Zr]Zr-Df'-GGSK-1/30 was investigated using breast tumor-bearing mice in vivo by PET/MRT imaging as well as by ex vivo organ biodistribution anal. Results: The mAb GGSK-1/30 stained specifically human breast tumor tissue and can be possibly used to predict the severity of disease progression based on the expression of the tumor-assocd. MUC1. For in vivo imaging, the Df'-conjugated mAb was radiolabeled with a radiochem. yield of 60 %, a radiochem. purity of 95 % and an apparent specific activity of 6.1 GBq/μmol. After 7 d, stabilities of 84 % in human serum and of 93 % in saline were obsd. In vitro cell studies showed strong binding to human tumor-assocd. MUC1 expressing breast cancer cells. The breast tumor-bearing mice showed an in vivo tumor uptake of >50 %ID/g and clearly visible specific enrichment of the radioconjugate via PET/MRT. Principal conclusions: Tumor-assocd. MUC1 is a very important biomarker for breast cancer next to the traditional markers estrogen receptor (ER), progesterone receptor (PR) and HER/2-neu. The mAb GGSK-1/30 can be used for the diagnosis of over 90% of breast cancers, including triple neg. breast cancer based on biopsy staining. Its radioimmunoconjugate represents a promising PET-tracer for breast cancer imaging selectively targeting breast cancer cells.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXlsVKmsbk%253D&md5=dbca95f8f30a8df92aec9a049a488e4c
    54. 54
      Festari, M. F.; Da Costa, V.; Rodríguez-Zraquia, S. A.; Costa, M.; Landeira, M.; Lores, P.; Solari-Saquieres, P.; Kramer, M. G.; Freire, T. The Tumor-Associated Tn Antigen Fosters Lung Metastasis and Recruitment of Regulatory T Cells in Triple Negative Breast Cancer. Glycobiology 2022, 32, 366– 379,  DOI: 10.1093/glycob/cwab123
      54
      The tumor-associated Tn antigen fosters lung metastasis and recruitment of regulatory T cells in triple negative breast cancer
      Festari, Maria Florencia; da Costa, Valeria; Rodriguez-Zraquia, Santiago A.; Costa, Monique; Landeira, Mercedes; Lores, Pablo; Solari-Saquieres, Patricia; Kramer, M. Gabriela; Freire, Teresa
      Glycobiology (2022), 32 (5), 366-379CODEN: GLYCE3; ISSN:1460-2423. (Oxford University Press)
      Cancer is a leading cause of death worldwide, accounting for nearly 10 million deaths. Among breast cancers (BC) subtypes, triple-neg. (TN) BC is characterized by metastatic progression and poor patient prognosis. Although, TNBC is initially sensitive to chemotherapy, many TNBC patients rapidly develop resistance, at which point metastatic disease is highly lethal. Cancer cells present phenotypic changes or mol. signatures that distinguish them from healthy cells. The Tn antigen (GalNAc-O-Thr/Ser), which constitutes a powerful tool as tumor marker, was recently reported to contribute to tumor growth. However, its role in BC-derived metastasis has not yet been addressed. In this work, we generated a pre-clin. orthotopic Tn+ model of metastatic TNBC, which mimics the patient surgical treatment and is useful to study the role of Tn in metastasis and immunoregulation. We obtained two different cell clones, which differed in their Tn antigen expression: a high Tn-expressing and a non-expressing clone. Interestingly, the Tn-pos. cell line generated significantly larger tumors and higher degree of lung metastases assocd. with a lower survival rate than the Tn-neg. and parental cell line. Furthermore, we also found that both tumors and draining-lymph nodes from Tn+-tumor-bearing mice presented a higher frequency of CD4+ FoxP3+ T cells, while their splenocytes expressed higher levels of IL-10. In conclusion, this work suggests that the Tn antigen participates in breast tumor growth and spreading, favoring metastases to the lungs that are assocd. with an immunoregulatory state, suggesting that Tn-based immunotherapy could be a strategy of choice to treat these tumors.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XisFKmtb3I&md5=66668ef3e2760c1ecacdea50dfbd8fc4
    55. 55
      Khosrowabadi, E.; Wenta, T.; Keskitalo, S.; Manninen, A.; Kellokumpu, S. Altered Glycosylation of Several Metastasis-Associated Glycoproteins with Terminal GalNAc Defines the Highly Invasive Cancer Cell Phenotype. Oncotarget 2022, 13, 73– 89,  DOI: 10.18632/oncotarget.28167
      55
      Altered glycosylation of several metastasis-associated glycoproteins with terminal GalNAc defines the highly invasive cancer cell phenotype
      Khosrowabadi Elham; Wenta Tomasz; Manninen Aki; Kellokumpu Sakari; Keskitalo Salla
      Oncotarget (2022), 13 (), 73-89 ISSN:.
      Several distinct metastasis-associated glycosylation changes have been shown to promote cancer cell invasion and metastasis, the main cause of death of cancer patients. However, it is unclear whether their presence reflects cell- or tissue-specific variations for metastasis, or species needed to drive different phases of the metastatic cascade. To address this issue from a different perspective, we investigated here whether different cancer cell lines share any glycotopes that are common and important for their invasive phenotype. By using lectin microarray glycan profiling and an established myoma tissue-based 3D invasion assay, we identified a single glycotope recognized by Helix Pomatia agglutinin (HPA), whose expression level in different cancer cells correlated significantly with their invasive potential. Lectin pull-down assay and LC-MS/MS analysis in highly- (A431 and SW-48) and poorly invasive (HepG2 and RCC4) cancer cells revealed ~85 glycoproteins of which several metastasis-promoting members of the integrin family of cell adhesion receptors, the epidermal growth factor receptor (EGFR) and the matrix metalloproteinase-14 (MMP-14) were among the abundant ones. Moreover, we showed that the level of the GalNAc glycotope in MMP-14, EGFR, αV-, β1- and β4 integrin in highly and poorly invasive cancer cells correlated positively with their invasive potential. Collectively, our findings suggest that altered glycosylation of several metastasis-associated glycoproteins with terminal GalNAc drives the highly invasive cancer cell phenotype.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB2M%252FotVagtQ%253D%253D&md5=f64cd9793dd2699928fc68aa5b362687
    56. 56
      Geijtenbeek, T. B. H.; Gringhuis, S. I. Signalling through C-Type Lectin Receptors: Shaping Immune Responses. Nat. Rev. Immunol. 2009, 9, 465– 479,  DOI: 10.1038/nri2569
      56
      Signalling through C-type lectin receptors: shaping immune responses
      Geijtenbeek, Teunis B. H.; Gringhuis, Sonja I.
      Nature Reviews Immunology (2009), 9 (7), 465-479CODEN: NRIABX; ISSN:1474-1733. (Nature Publishing Group)
      A review. C-type lectin receptors (CLRs) expressed by dendritic cells are crucial for tailoring immune responses to pathogens. Following pathogen binding, CLRs trigger distinct signalling pathways that induce the expression of specific cytokines which det. T cell polarization fates. Some CLRs can induce signalling pathways that directly activate nuclear factor-κB, whereas other CLRs affect signalling by Toll-like receptors. Dissecting these signalling pathways and their effects on host immune cells is essential to understand the mol. mechanisms involved in the induction of adaptive immune responses. In this Review we describe the role of CLR signalling in regulating adaptive immunity and immunopathogenesis and discuss how this knowledge can be harnessed for the development of innovative vaccination approaches.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXntFCgurc%253D&md5=91f3810595e1bb4794efa63a97321ecf

  • Supporting Information

    Supporting Information

    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/jacs.2c12843.

    • Experimental details and characterization of the synthetic antigens (PDF)


    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.