Allo-Specific Humoral Responses: New Methods for Screening Donor-Specific Antibody and Characterization of HLA-Specific Memory B Cells

Front Immunol. 2021 Jul 13;12:705140. doi: 10.3389/fimmu.2021.705140. eCollection 2021.


Antibody-mediated allograft rejection (AMR) causes more kidney transplant failure than any other single cause. AMR is mediated by antibodies recognizing antigens expressed by the graft, and antibodies generated against major histocompatibility complex (MHC) mismatches are especially problematic. Most research directed towards the management of clinical AMR has focused on identifying and characterizing circulating donor-specific HLA antibody (DSA) and optimizing therapies that reduce B-cell activation and/or block antibody secretion by inhibiting plasmacyte survival. Here we describe a novel set of reagents and techniques to allow more specific measurements of MHC sensitization across different animal transplant models. Additionally, we have used these approaches to isolate and clone individual HLA-specific B cells from patients sensitized by pregnancy or transplantation. We have identified and characterized the phenotypes of individual HLA-specific B cells, determined the V(D)J rearrangements of their paired H and L chains, and generated recombinant antibodies to determine affinity and specificity. Knowledge of the BCR genes of individual HLA-specific B cells will allow identification of clonally related B cells by high-throughput sequence analysis of peripheral blood mononuclear cells and permit us to re-construct the origins of HLA-specific B cells and follow their somatic evolution by mutation and selection.

Keywords: B cell; antibody mediated rejection (AMR); donor specific antibody (DSA); sensitization; transplantation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibody Specificity
  • B-Lymphocyte Subsets / immunology*
  • Cell Line
  • Cells, Cultured
  • Clonal Evolution
  • Clone Cells
  • Female
  • Gene Rearrangement, B-Lymphocyte
  • Genes, Reporter
  • Graft Rejection / immunology*
  • HLA Antigens / immunology*
  • Histocompatibility Antigens / biosynthesis
  • Histocompatibility Antigens / immunology
  • Humans
  • Immunoglobulin G / immunology
  • Immunologic Memory*
  • Indicators and Reagents
  • Isoantibodies / blood*
  • Isoantibodies / immunology
  • Lymphocyte Activation
  • Macaca mulatta
  • Mice
  • Mice, Inbred C57BL
  • Models, Animal
  • RNA, Guide, Kinetoplastida / genetics
  • Skin Transplantation
  • Species Specificity
  • Specific Pathogen-Free Organisms
  • V(D)J Recombination
  • beta 2-Microglobulin / antagonists & inhibitors
  • beta 2-Microglobulin / genetics


  • HLA Antigens
  • Histocompatibility Antigens
  • Immunoglobulin G
  • Indicators and Reagents
  • Isoantibodies
  • RNA, Guide
  • beta 2-Microglobulin