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Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor

There is far less information available about the tumor infiltrating B (TIL-B) cells, than about the tumor infiltrating T cells. We focused on discovering the features and potential role of B lymphocytes in solid tumors. Our project aimed to develop innovative strategies to define cancer membrane st...

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Autores principales: Kotlan, Beatrix, Horvath, Szabolcs, Eles, Klara, Plotar, Vanda K., Naszados, Gyorgy, Czirbesz, Katalin, Blank, Miri, Farkas, Emil, Toth, Laszlo, Tovari, Jozsef, Szekacs, Andras, Shoenfeld, Yehuda, Godeny, Maria, Kasler, Miklos, Liszkay, Gabriella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6459966/
https://www.ncbi.nlm.nih.gov/pubmed/31024530
http://dx.doi.org/10.3389/fimmu.2019.00650
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author Kotlan, Beatrix
Horvath, Szabolcs
Eles, Klara
Plotar, Vanda K.
Naszados, Gyorgy
Czirbesz, Katalin
Blank, Miri
Farkas, Emil
Toth, Laszlo
Tovari, Jozsef
Szekacs, Andras
Shoenfeld, Yehuda
Godeny, Maria
Kasler, Miklos
Liszkay, Gabriella
author_facet Kotlan, Beatrix
Horvath, Szabolcs
Eles, Klara
Plotar, Vanda K.
Naszados, Gyorgy
Czirbesz, Katalin
Blank, Miri
Farkas, Emil
Toth, Laszlo
Tovari, Jozsef
Szekacs, Andras
Shoenfeld, Yehuda
Godeny, Maria
Kasler, Miklos
Liszkay, Gabriella
author_sort Kotlan, Beatrix
collection PubMed
description There is far less information available about the tumor infiltrating B (TIL-B) cells, than about the tumor infiltrating T cells. We focused on discovering the features and potential role of B lymphocytes in solid tumors. Our project aimed to develop innovative strategies to define cancer membrane structures. We chose two solid tumor types, with variable to considerable B cell infiltration. The strategy we set up with invasive breast carcinoma, showing medullary features, has been introduced and standardized in metastatic melanoma. After detecting B lymphocytes by immunohistochemistry, VH-JH, Vκ-Jκ immunoglobulin rearranged V region genes were amplified by RT-PCR, from TIL-B cDNA. Immunoglobulin variable-region genes of interest were cloned, sequenced, and subjected to a comparative DNA analysis. Single-chain variable (scFv) antibody construction was performed in selected cases to generate a scFv library and to test tumor binding capacity. DNA sequence analysis revealed an overrepresented VH3-1 cluster, represented both in the breast cancer and the melanoma TIL-B immunoglobulin repertoire. We observed that our previously defined anti GD3 ganglioside-binder antibody-variable region genes were present in melanoma as well. Our antibody fragments showed binding potential to disialylated glycosphingolipids (GD3 ganglioside) and their O acetylated forms on melanoma cancer cells. We conclude that our results have a considerable tumor immunological impact, as they reveal the power of TIL-B cells to recognize strong tumor-associated glycosphingolipid structures on melanomas and other solid tumors. As tumor-derived gangliosides affect immune cell functions and reduce the B lymphocytes' antibody production, we suspect an important B lymphocyte and cancer cell crosstalk mechanism. We not only described the isolation and specificity testing of the tumor infiltrating B cells, but also showed the TIL-B cells' highly tumor-associated GD3 ganglioside-revealing potential in melanomas. The present data help to identify new cancer-associated biomarkers that may serve for novel cancer diagnostics. The two-direction regulation mechanism between immune B cells and the tumor could eventually be developed into an innovative cancer treatment strategy.
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spelling pubmed-64599662019-04-25 Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor Kotlan, Beatrix Horvath, Szabolcs Eles, Klara Plotar, Vanda K. Naszados, Gyorgy Czirbesz, Katalin Blank, Miri Farkas, Emil Toth, Laszlo Tovari, Jozsef Szekacs, Andras Shoenfeld, Yehuda Godeny, Maria Kasler, Miklos Liszkay, Gabriella Front Immunol Immunology There is far less information available about the tumor infiltrating B (TIL-B) cells, than about the tumor infiltrating T cells. We focused on discovering the features and potential role of B lymphocytes in solid tumors. Our project aimed to develop innovative strategies to define cancer membrane structures. We chose two solid tumor types, with variable to considerable B cell infiltration. The strategy we set up with invasive breast carcinoma, showing medullary features, has been introduced and standardized in metastatic melanoma. After detecting B lymphocytes by immunohistochemistry, VH-JH, Vκ-Jκ immunoglobulin rearranged V region genes were amplified by RT-PCR, from TIL-B cDNA. Immunoglobulin variable-region genes of interest were cloned, sequenced, and subjected to a comparative DNA analysis. Single-chain variable (scFv) antibody construction was performed in selected cases to generate a scFv library and to test tumor binding capacity. DNA sequence analysis revealed an overrepresented VH3-1 cluster, represented both in the breast cancer and the melanoma TIL-B immunoglobulin repertoire. We observed that our previously defined anti GD3 ganglioside-binder antibody-variable region genes were present in melanoma as well. Our antibody fragments showed binding potential to disialylated glycosphingolipids (GD3 ganglioside) and their O acetylated forms on melanoma cancer cells. We conclude that our results have a considerable tumor immunological impact, as they reveal the power of TIL-B cells to recognize strong tumor-associated glycosphingolipid structures on melanomas and other solid tumors. As tumor-derived gangliosides affect immune cell functions and reduce the B lymphocytes' antibody production, we suspect an important B lymphocyte and cancer cell crosstalk mechanism. We not only described the isolation and specificity testing of the tumor infiltrating B cells, but also showed the TIL-B cells' highly tumor-associated GD3 ganglioside-revealing potential in melanomas. The present data help to identify new cancer-associated biomarkers that may serve for novel cancer diagnostics. The two-direction regulation mechanism between immune B cells and the tumor could eventually be developed into an innovative cancer treatment strategy. Frontiers Media S.A. 2019-04-05 /pmc/articles/PMC6459966/ /pubmed/31024530 http://dx.doi.org/10.3389/fimmu.2019.00650 Text en Copyright © 2019 Kotlan, Horvath, Eles, Plotar, Naszados, Czirbesz, Blank, Farkas, Toth, Tovari, Szekacs, Shoenfeld, Godeny, Kasler and Liszkay. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Kotlan, Beatrix
Horvath, Szabolcs
Eles, Klara
Plotar, Vanda K.
Naszados, Gyorgy
Czirbesz, Katalin
Blank, Miri
Farkas, Emil
Toth, Laszlo
Tovari, Jozsef
Szekacs, Andras
Shoenfeld, Yehuda
Godeny, Maria
Kasler, Miklos
Liszkay, Gabriella
Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title_full Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title_fullStr Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title_full_unstemmed Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title_short Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor
title_sort tumor-associated disialylated glycosphingolipid antigen-revealing antibodies found in melanoma patients' immunoglobulin repertoire suggest a two-direction regulation mechanism between immune b cells and the tumor
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6459966/
https://www.ncbi.nlm.nih.gov/pubmed/31024530
http://dx.doi.org/10.3389/fimmu.2019.00650
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