Article ; Online: MHC Tetramers Specifically Identify High- and Low-avidity Donor-specific B Cells in Transplantation Tolerance and Rejection.
2023 Volume 107, Issue 12, Page(s) 2526–2532
Abstract: Background: Although donor-specific antibody pre- and posttransplantation is routinely assessed, accurate quantification of memory alloreactive B cells that mediate recall antibody response remains challenging. Major histocompatibility complex (MHC) ... ...
| Abstract | Background: Although donor-specific antibody pre- and posttransplantation is routinely assessed, accurate quantification of memory alloreactive B cells that mediate recall antibody response remains challenging. Major histocompatibility complex (MHC) tetramers have been used to identify alloreactive B cells in mice and humans, but the specificity of this approach has not been rigorously assessed. Methods: B-cell receptors from MHC tetramer-binding single B cells were expressed as mouse recombinant immunoglobulin G1 (rIgG1) monoclonal antibodies, and the specificity was assessed with a multiplex bead assay. Relative binding avidity of rIgG1 was measured by modified dilution series technique and surface plasmon resonance. Additionally, immunoglobulin heavy chain variable regions of 50 individual B-cell receptors were sequenced to analyze the rate of somatic hypermutation. Results: The multiplex bead assay confirmed that expressed rIgG1 monoclonal antibodies were preferentially bound to bait MHC class II I-E d over control I-A d and I-A b tetramers. Furthermore, the dissociation constant 50 binding avidities of the rIgG1 ranged from 10 mM to 7 nM. The majority of tetramer-binding B cells were low avidity, and ~12.8% to 15.2% from naive and tolerant mice and 30.9% from acute rejecting mice were higher avidity (dissociation constant 50 <1 mM). Conclusions: Collectively, these studies demonstrate that donor MHC tetramers, under stringent binding conditions with decoy self-MHC tetramers, can specifically identify a broad repertoire of donor-specific B cells under conditions of rejection and tolerance. |
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| MeSH term(s) | Humans ; Mice ; Animals ; Transplantation Tolerance ; Major Histocompatibility Complex ; Histocompatibility Antigens Class II ; Immunoglobulin G ; Antibodies, Monoclonal ; Receptors, Antigen, B-Cell |
| Chemical Substances | Histocompatibility Antigens Class II ; Immunoglobulin G ; Antibodies, Monoclonal ; Receptors, Antigen, B-Cell |
| Language | English |
| Publishing date | 2023-11-22 |
| Publishing country | United States |
| Document type | Journal Article |
| ZDB-ID | 208424-7 |
| ISSN | 1534-6080 ; 0041-1337 |
| ISSN (online) | 1534-6080 |
| ISSN | 0041-1337 |
| DOI | 10.1097/TP.0000000000004702 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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