In vitro selection of lymphocytic choriomeningitis virus escape mutants by cytotoxic T lymphocytes

Toni Aebischer, Demetrius Moskophidis, Urs Hoffmann Rohrer, Rolf M. Zinkernagel, Hans Hengartner

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Cytotoxic T lymphocyte (CTL)-mediated cytolysis is induced via the interaction of the specific T-cell antigen receptor and the peptidic viral antigen associated with the major histocompatibility complex class I antigen. Here we demonstrate in vitro that lymphocytic choriomeningitis virus (LCMV) can escape the cytotoxic activity of LCMV-specific cloned CTLs by single amino acid changes within the recognized T-cell epitope defined by residues 275-289 of the LCMV glycoprotein [LCMV-GP-(275-289)]. LCMV-infected fibroblasts at a multiplicity of infection of 10-3 exposed to virus-specific CTL at an effector-to-target cell ratio of 4:14 hr after infection was optimal for virus mutant selection. The selections were carried out with three LCMV-GP-(275-289)-specific CTL clones expressing T-cell antigen receptors containing the identical variable gene segments Vα4 and Vβ10 but different junctional regions; selection was also possible with LCMV-GP(275-289)-specific cytotoxic polyclonal T cells. The most common escape mutation was an amino acid change of asparagine (AAT) to aspartic acid (GAT) at position 280; an additional mutation was glycine (GGT) to aspartic acid (GAT) at position 282. The results presented show that relevant point mutations within the T-cell epitope of LCMV-GP-(275-289) occur frequently and that they are selectable in vitro by CTLs.

Original languageEnglish (US)
Pages (from-to)11047-11051
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number24
DOIs
StatePublished - Jan 1 1991

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Lymphocytic choriomeningitis virus
Cytotoxic T-Lymphocytes
Aspartic Acid
T-Lymphocyte Epitopes
T-Cell Antigen Receptor
Viruses
Amino Acids
Histocompatibility Antigens Class I
Mutation
In Vitro Techniques
Viral Antigens
Asparagine
Infection
Major Histocompatibility Complex
Point Mutation
Glycine
Glycoproteins
Clone Cells
Fibroblasts
T-Lymphocytes

Keywords

  • T-cell epitope
  • Virus mutants
  • Virus selection

ASJC Scopus subject areas

  • General

Cite this

In vitro selection of lymphocytic choriomeningitis virus escape mutants by cytotoxic T lymphocytes. / Aebischer, Toni; Moskophidis, Demetrius; Rohrer, Urs Hoffmann; Zinkernagel, Rolf M.; Hengartner, Hans.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 24, 01.01.1991, p. 11047-11051.

Research output: Contribution to journalArticle

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