Induction and exhaustion of lymphocytic choriomeningitis virus-specific cytotoxic T lymphocytes visualized using soluble tetrameric major histocompatibility complex class I-peptide complexes

J Exp Med. 1998 May 4;187(9):1383-93. doi: 10.1084/jem.187.9.1383.

Abstract

This study describes the construction of soluble major histocompatibility complexes consisting of the mouse class I molecule, H-2Db, chemically biotinylated beta2 microglobulin and a peptide epitope derived from the glycoprotein (GP; amino acids 33-41) of lymphocytic choriomeningitis virus (LCMV). Tetrameric class I complexes, which were produced by mixing the class I complexes with phycoerythrin-labeled neutravidin, permitted direct analysis of virus-specific cytotoxic T lymphocytes (CTLs) by flow cytometry. This technique was validated by (a) staining CD8+ cells in the spleens of transgenic mice that express a T cell receptor (TCR) specific for H-2Db in association with peptide GP33-41, and (b) by staining virus-specific CTLs in the cerebrospinal fluid of C57BL/6 (B6) mice that had been infected intracranially with LCMV-DOCILE. Staining of spleen cells isolated from B6 mice revealed that up to 40% of CD8(+) T cells were GP33 tetramer+ during the initial phase of LCMV infection. In contrast, GP33 tetramers did not stain CD8+ T cells isolated from the spleens of B6 mice that had been infected 2 mo previously with LCMV above the background levels found in naive mice. The fate of virus-specific CTLs was analyzed during the acute phase of infection in mice challenged both intracranially and intravenously with a high or low dose of LCMV-DOCILE. The results of the study show that the outcome of infection by LCMV is determined by antigen load alone. Furthermore, the data indicate that deletion of virus-specific CTLs in the presence of excessive antigen is preceded by TCR downregulation and is dependent upon perforin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD8-Positive T-Lymphocytes / immunology
  • Flow Cytometry
  • Glycoproteins / immunology
  • Histocompatibility Antigens Class I / immunology
  • Interferon-gamma / analysis
  • Lymphocytic choriomeningitis virus / chemistry
  • Lymphocytic choriomeningitis virus / immunology*
  • Major Histocompatibility Complex / immunology*
  • Membrane Glycoproteins / deficiency
  • Mice
  • Mice, Transgenic
  • Peptides / immunology
  • Perforin
  • Pore Forming Cytotoxic Proteins
  • Protein Conformation
  • Receptors, Antigen, T-Cell / physiology
  • Spleen / immunology
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / virology*
  • beta 2-Microglobulin / metabolism

Substances

  • Glycoproteins
  • Histocompatibility Antigens Class I
  • Membrane Glycoproteins
  • Peptides
  • Pore Forming Cytotoxic Proteins
  • Receptors, Antigen, T-Cell
  • beta 2-Microglobulin
  • Perforin
  • Interferon-gamma