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A Memory B Cell Crossmatch Assay for Quantification of Donor‐Specific Memory B Cells in the Peripheral Blood of HLA ‐Immunized Individuals
Author(s) -
Karahan G. E.,
Vaal Y. J. H.,
Krop J.,
Wehmeier C.,
Roelen D. L.,
Claas F. H. J.,
Heidt S.
Publication year - 2017
Publication title -
american journal of transplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.89
H-Index - 188
eISSN - 1600-6143
pISSN - 1600-6135
DOI - 10.1111/ajt.14293
Subject(s) - elispot , immunology , human leukocyte antigen , memory b cell , antibody , medicine , antigen , b cell , b 1 cell , biology , immune system , antigen presenting cell , t cell , cd8
Humoral responses against mismatched donor HLA are routinely measured as serum HLA antibodies, which are mainly produced by bone marrow–residing plasma cells. Individuals with a history of alloimmunization but lacking serum antibodies may harbor circulating dormant memory B cells, which may rapidly become plasma cells on antigen reencounter. Currently available methods to detect HLA ‐specific memory B cells are scarce and insufficient in quantifying the complete donor‐specific memory B cell response due to their dependence on synthetic HLA molecules. We present a highly sensitive and specific tool for quantifying donor‐specific memory B cells in peripheral blood of individuals using cell lysates covering the complete HLA class I and class II repertoire of an individual. Using this enzyme‐linked immunospot ( ELISpot) assay, we found a median frequency of 31 HLA class I and 89 HLA class II –specific memory B cells per million IgG‐producing cells directed at paternal HLA in peripheral blood samples from women (n = 22) with a history of pregnancy, using cell lysates from spouses. The donor‐specific memory B cell ELISpot can be used in HLA diagnostic laboratories as a cross‐match assay to quantify donor‐specific memory B cells in patients with a history of sensitizing events.