Oligoclonal expansion of major histocompatibility complex class I-restricted cytolytic T lymphocytes during a primary immune response in vivo: direct monitoring by flow cytometry and polymerase chain reaction

J Exp Med. 1993 May 1;177(5):1487-92. doi: 10.1084/jem.177.5.1487.


Previous T cell receptor (TCR) sequence analysis of a panel of 23 H-2Kd restricted cytotoxic T lymphocyte (CTL) clones recognizing the decapeptide HLA-CW3 170-179 revealed a striking conservation of TCR structure, in that all clones examined used V beta 10 and J alpha pHDS58 segments. We show here that the primary response in vivo after intraperitoneal injection of DBA/2 mice with HLA-CW3 expressing transfectants of syngeneic P815 (H-2d) tumor cells is characterized by a dramatic expansion of CD8+ V beta 10+ CTL in the peritoneal cavity and draining (mesenteric) lymph node, as well as in peripheral blood. Additional analysis of TCR on HLA-CW3 immune populations by flow cytometry and polymerase chain reaction further indicates that the vast majority of responding CD8+ cells express restricted V alpha domains, a dominant J alpha segment (pHDS58), and a conserved CDR3 length for both alpha and beta chains. This novel system provides a unique opportunity to directly monitor an oligoclonal primary antigen specific immune response in vivo at the single cell level independently of functional assays.

MeSH terms

  • Animals
  • CD8 Antigens
  • Cells, Cultured
  • Clone Cells
  • Female
  • Flow Cytometry
  • HLA-C Antigens / immunology
  • Histocompatibility Antigens Class I / immunology*
  • Mice
  • Mice, Inbred DBA
  • Peptide Fragments / immunology
  • Polymerase Chain Reaction
  • Receptors, Antigen, T-Cell, alpha-beta
  • T-Lymphocytes, Cytotoxic / cytology
  • T-Lymphocytes, Cytotoxic / immunology*


  • CD8 Antigens
  • HLA-C Antigens
  • HLA-C*03 antigen
  • Histocompatibility Antigens Class I
  • Peptide Fragments
  • Receptors, Antigen, T-Cell, alpha-beta