T-cell hybridomas from HLA-transgenic mice as tools for analysis of human antigen processing

J Immunol Methods. 2003 Oct 1;281(1-2):129-42. doi: 10.1016/j.jim.2003.07.004.

Abstract

The study of antigen processing and presentation by human antigen presenting cells (APC) has been limited by difficulties of producing and maintaining human T-cell clones. Murine T-cell hybridomas have advantages for detecting specific peptide-MHC complexes on APC. Human antigen-specific immortalized T-cell lines have not been successfully produced. We report and validate the use of transgenic mice with human MHC genes for HLA-A2, DR1 and DR4 to produce murine T-cell hybridomas that are restricted to human HLA alleles and respond to human macrophages, dendritic cells (DC), and B-cell lines. Hybridomas restricted by human MHC-I and -II specific for influenza matrix protein, tetanus toxoid, diphtheria antigen CRM(197), and various M. tuberculosis antigens were produced. Epitope specificity was determined for several hybridomas. T hybridomas recognized peptide-MHC complexes on fixed APC for analysis of kinetics or susceptibility to inhibitors of antigen processing. T hybridomas restricted by human MHC represent convenient and powerful tools for the study of antigen processing by human APC.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigen Presentation*
  • Antigen-Presenting Cells / physiology
  • Cell Line
  • Epitope Mapping
  • HLA-A2 Antigen / physiology*
  • Humans
  • Hybridomas / immunology*
  • Interferon-gamma / pharmacology
  • Mice
  • Mice, Transgenic
  • Molecular Sequence Data
  • T-Lymphocytes / immunology*

Substances

  • HLA-A2 Antigen
  • Interferon-gamma