The human T cell receptor V beta repertoire of normal peripheral blood lymphocytes before and after mitogen stimulation

Clin Exp Immunol. 1993 May;92(2):361-6. doi: 10.1111/j.1365-2249.1993.tb03405.x.

Abstract

Mitogen stimulation of T cells in vitro has been employed in the analysis of the T cell antigen receptor (TCR) repertoire and as a method of generating T cell lines and clones. It has been suspected for some time that mitogen stimulation may bias the repertoire. We have addressed this problem employing a semi-quantitative technique utilizing the polymerase chain reaction (PCR) and flow cytometry. Using this PCR method and a panel of primers to 22 V beta subgroups, the V beta repertoire of both unstimulated and phytohaemagglutinin (PHA)-stimulated peripheral T cells from eight healthy individuals was investigated. The samples were also analysed by flow cytometry using anti-V beta 2, V beta 5 and V beta 8 MoAbs. A significant increase in the expression of V beta 6, V beta 7.2 and V beta 10.1 was found in all eight samples of PHA-stimulated T cells compared with unstimulated T cells using the PCR method. In contrast, no differences were found between unstimulated and PHA-stimulated T cells by flow cytometry. These results question the validity of using mitogen-stimulated T cells to investigate TCR gene usage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • CD3 Complex / analysis
  • Flow Cytometry
  • Gene Rearrangement, beta-Chain T-Cell Antigen Receptor*
  • Humans
  • Lymphocyte Activation*
  • Mitogens / pharmacology
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / chemistry
  • Polymerase Chain Reaction
  • Receptors, Antigen, T-Cell, alpha-beta / genetics*
  • T-Lymphocytes / physiology*

Substances

  • CD3 Complex
  • Mitogens
  • Oligodeoxyribonucleotides
  • Receptors, Antigen, T-Cell, alpha-beta