Visualizing interleukin 2 gene expression at the single cell level

Eur J Immunol. 1989 Sep;19(9):1619-24. doi: 10.1002/eji.1830190915.

Abstract

To analyze the expression of the interleukin (IL)2 gene at the single cell level, we have constructed a chimeric gene in which the regulatory sequences from the mouse IL2 gene were fused 5' proximal to the coding region of the Escherichia coli beta-galactosidase (lacZ) gene. Once stably introduced into a T cell hybridoma, the IL 2-lacZ reporter gene was shown to display a pattern of induction similar to the one observed for the resident IL 2 gene. Two points emerged from monitoring IL 2 expression for the resident IL2 gene. Two points emerged from monitoring IL 2 expression at the single cell level. First, upon activation the expression of the IL2-lacZ gene appears asynchronous. Second, at the peak of induction the percentage of beta-galactosidase+ cells never reached 100% and the level of beta-galactosidase reached among positive cells was highly heterogeneous within a cloned population of T cells. In combination with the possibility to sort viable lymphocytes according to lacZ expression, the IL2-lacZ reporter gene described herein should provide a way to isolate somatic cell mutants deficient in signal transduction by the T cell antigen receptor complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens
  • Cloning, Molecular
  • Flow Cytometry
  • Gene Expression Regulation
  • Hybridomas
  • Interleukin-2 / genetics*
  • Lymphocyte Activation
  • Mice
  • Promoter Regions, Genetic
  • Receptors, Antigen, T-Cell / physiology*
  • Recombinant Fusion Proteins / genetics
  • T-Lymphocytes / physiology*
  • Transfection

Substances

  • Antigens
  • Interleukin-2
  • Receptors, Antigen, T-Cell
  • Recombinant Fusion Proteins