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Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR

In kidney transplantation, eplet mismatches between donor and recipient have been associated with de novo donor‐specific antibody development. Eplets are theoretically defined configurations of polymorphic amino acids and require experimental verification to establish whether they can be bound by al...

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Autores principales: Kramer, Cynthia S. M., Franke‐van Dijk, Marry E. I., Bakker, Kim H., Uyar‐Mercankaya, Merve, Karahan, Gonca E., Roelen, Dave L., Claas, Frans H. J., Heidt, Sebastiaan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7754395/
https://www.ncbi.nlm.nih.gov/pubmed/32342632
http://dx.doi.org/10.1111/ajt.15950
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author Kramer, Cynthia S. M.
Franke‐van Dijk, Marry E. I.
Bakker, Kim H.
Uyar‐Mercankaya, Merve
Karahan, Gonca E.
Roelen, Dave L.
Claas, Frans H. J.
Heidt, Sebastiaan
author_facet Kramer, Cynthia S. M.
Franke‐van Dijk, Marry E. I.
Bakker, Kim H.
Uyar‐Mercankaya, Merve
Karahan, Gonca E.
Roelen, Dave L.
Claas, Frans H. J.
Heidt, Sebastiaan
author_sort Kramer, Cynthia S. M.
collection PubMed
description In kidney transplantation, eplet mismatches between donor and recipient have been associated with de novo donor‐specific antibody development. Eplets are theoretically defined configurations of polymorphic amino acids and require experimental verification to establish whether they can be bound by alloantibodies. Human HLA‐specific monoclonal antibodies (mAbs) have been instrumental for this purpose but are largely lacking for HLA class II. In this study, we isolated single HLA‐DR‐specific memory B cells from peripheral blood of immunized individuals (n = 3) using HLA class II tetramers to generate recombinant human HLA‐DR antigen‐reactive mAbs (n = 5). Comparison of the amino acid composition of the reactive HLA alleles in relation to the antibody reactivity patterns led to identification of 3 configurations, 70Q 73A, 31F 32Y 37Y, and 14K 25Q recognized, respectively, by HLA‐DRB1*01:01, HLA‐DRB1*04:01, and HLA‐DRB1*07:01 antigen‐reactive mAbs. The first 2 correspond to eplets 70QA and 31FYY and can now be considered antibody verified. The latter indicates that eplet 25Q needs to be redefined before being considered as antibody verified. Generation and reactivity analysis of human HLA‐DR mAbs allowed for identification of amino acid configurations corresponding to known eplets, whereas the other patterns may be used to redefine eplets with similar, but not identical predicted amino acid composition.
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spelling pubmed-77543952020-12-23 Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR Kramer, Cynthia S. M. Franke‐van Dijk, Marry E. I. Bakker, Kim H. Uyar‐Mercankaya, Merve Karahan, Gonca E. Roelen, Dave L. Claas, Frans H. J. Heidt, Sebastiaan Am J Transplant ORIGINAL ARTICLES In kidney transplantation, eplet mismatches between donor and recipient have been associated with de novo donor‐specific antibody development. Eplets are theoretically defined configurations of polymorphic amino acids and require experimental verification to establish whether they can be bound by alloantibodies. Human HLA‐specific monoclonal antibodies (mAbs) have been instrumental for this purpose but are largely lacking for HLA class II. In this study, we isolated single HLA‐DR‐specific memory B cells from peripheral blood of immunized individuals (n = 3) using HLA class II tetramers to generate recombinant human HLA‐DR antigen‐reactive mAbs (n = 5). Comparison of the amino acid composition of the reactive HLA alleles in relation to the antibody reactivity patterns led to identification of 3 configurations, 70Q 73A, 31F 32Y 37Y, and 14K 25Q recognized, respectively, by HLA‐DRB1*01:01, HLA‐DRB1*04:01, and HLA‐DRB1*07:01 antigen‐reactive mAbs. The first 2 correspond to eplets 70QA and 31FYY and can now be considered antibody verified. The latter indicates that eplet 25Q needs to be redefined before being considered as antibody verified. Generation and reactivity analysis of human HLA‐DR mAbs allowed for identification of amino acid configurations corresponding to known eplets, whereas the other patterns may be used to redefine eplets with similar, but not identical predicted amino acid composition. John Wiley and Sons Inc. 2020-05-15 2020-12 /pmc/articles/PMC7754395/ /pubmed/32342632 http://dx.doi.org/10.1111/ajt.15950 Text en © 2020 The Authors. American Journal of Transplantation published by Wiley Periodicals LLC on behalf of The American Society of Transplantation and the American Society of Transplant Surgeons This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle ORIGINAL ARTICLES
Kramer, Cynthia S. M.
Franke‐van Dijk, Marry E. I.
Bakker, Kim H.
Uyar‐Mercankaya, Merve
Karahan, Gonca E.
Roelen, Dave L.
Claas, Frans H. J.
Heidt, Sebastiaan
Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title_full Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title_fullStr Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title_full_unstemmed Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title_short Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA‐DR
title_sort generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on hla‐dr
topic ORIGINAL ARTICLES
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7754395/
https://www.ncbi.nlm.nih.gov/pubmed/32342632
http://dx.doi.org/10.1111/ajt.15950
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