Identification, isolation, and culture of HLA-A2-specific B lymphocytes using MHC class I tetramers

J Immunol. 2003 Dec 15;171(12):6599-603. doi: 10.4049/jimmunol.171.12.6599.

Abstract

Characterizing the individual B cells that participate in the production of anti-HLA Abs requires isolation and culture of these cells and a suitable assay for detection of Abs produced in these B cell cultures. We previously showed that B cell precursors, programmed for anti-HLA Ab secretion, are present at measurable frequencies in peripheral blood of women immunized by pregnancy. In this study, we show that tetrameric HLA-A2, although designed for characterization of CTLs, provides a suitable affinity ligand for isolation of allospecific B cells, which subsequently can be induced to produce HLA-A2 Ab in a CD40-driven culture system. The validity of this concept was established by assaying human hybridomas, producing anti-HLA Abs, for specific tetrameric HLA-A2 binding. The availability of anti-HLA Ab-producing B cell cultures that are established without immortalization will be of value when T-B cell interaction is studied at an alloantigen-specific level.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / blood
  • B-Lymphocyte Subsets / chemistry
  • B-Lymphocyte Subsets / immunology*
  • B-Lymphocyte Subsets / metabolism*
  • Cell Culture Techniques / methods
  • Cell Separation / methods
  • Cells, Cultured
  • Epitopes, B-Lymphocyte / analysis
  • Epitopes, B-Lymphocyte / immunology*
  • Female
  • Flow Cytometry
  • HLA-A2 Antigen / analysis
  • HLA-A2 Antigen / immunology*
  • Histocompatibility Antigens Class I / analysis
  • Histocompatibility Antigens Class I / immunology
  • Humans
  • Hybridomas
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / blood
  • Immunoglobulin M / biosynthesis
  • Immunoglobulin M / blood
  • Male
  • Oligopeptides / immunology*
  • Staining and Labeling

Substances

  • Antibodies, Monoclonal
  • Epitopes, B-Lymphocyte
  • HLA-A2 Antigen
  • Histocompatibility Antigens Class I
  • Immunoglobulin G
  • Immunoglobulin M
  • Oligopeptides