T cell lines specific for a synthetic Heymann nephritis peptide derived from the receptor-associated protein

Clin Exp Immunol. 2000 Jul;121(1):157-64. doi: 10.1046/j.1365-2249.2000.01262.x.

Abstract

Pathogenic antigens involved in the induction of Heymann nephritis (HN), an experimental rat model of human membranous nephritis, have been identified in megalin (gp330) and the receptor-associated protein (RAP) [1,2]. A pathogenic epitope has been identified in RAP (amino acid 1-86) that plays a significant role in the formation of immune deposits in glomeruli in HN. A synthetic peptide (P31-53) derived from RAP1-86 contains a pathogenic epitope recognized by antibodies eluted from glomerular immune deposits and includes two putative RT-1B1 MHC class II-binding motifs. We have investigated whether RAP P31-53 can be recognized by T cells. Five peptide-specific T cell lines were generated from regional lymph node (LN) T cells from Lewis rats immunized with P31-53. The T cell lines were characterized by using a T cell proliferation assay for their specificity, FACS and MHC restriction assay for the phenotype, reverse transcription-polymerase chain reaction for TCR Vbeta repertoire and cytokine expression, and cloning and sequencing for the analysis of the CDR3 sequence of TCR. The helper function of the T cell line was confirmed by autoantibody production in vitro. In this study, we clearly identify that the synthetic pathogenic peptide P31-53 contains a T cell epitope recognized by CD4+ Th2 cells in Lewis rats. This recognition was restricted by MHC class II RT1.B1. These CD4+ Th2 cells were able to promote B cells to produce specific antibodies and used a restricted set of TCR Vbeta genes with preferential usage of Vbeta18. A charged amino acid motif at the CDR3 region of predominant TCR Vbeta subfamilies may contribute to the specific ability of these cells to recognize the immunogenic T cell epitope within RAP peptide P31-53.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoantibodies / biosynthesis
  • Autoantigens / immunology*
  • B-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / immunology
  • CD8 Antigens / immunology
  • Cell Division
  • Cell Line
  • Cytokines / genetics
  • Glomerulonephritis / immunology*
  • Heymann Nephritis Antigenic Complex
  • Histocompatibility Antigens / immunology
  • Humans
  • Lymph Nodes / cytology
  • Lymphocyte Activation / immunology
  • Male
  • Membrane Glycoproteins / immunology*
  • Molecular Sequence Data
  • Peptides / chemical synthesis
  • Peptides / immunology
  • Rats
  • Rats, Inbred Lew
  • Receptors, Antigen, T-Cell, alpha-beta / genetics
  • Receptors, Antigen, T-Cell, alpha-beta / immunology
  • T-Lymphocytes / immunology*
  • Th2 Cells / immunology

Substances

  • Autoantibodies
  • Autoantigens
  • CD8 Antigens
  • Cytokines
  • Heymann Nephritis Antigenic Complex
  • Histocompatibility Antigens
  • Membrane Glycoproteins
  • Peptides
  • Receptors, Antigen, T-Cell, alpha-beta
  • histocompatibility antigens RT, rat