Visualizing interleukin 2 gene expression at the single cell level.
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.
Emilie
D
D
Centre d'Immunologie INSERM-CNRS de Marseille-Luminy, France.
Peuchmaur
M
M
Barad
M
M
Jouin
H
H
Maillot
M C
MC
Couez
D
D
Nicolas
J F
JF
Malissen
B
B
D000818:Animals; D000941:Antigens; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006825:Hybridomas; D007376:Interleukin-2 / Q000235:genetics*; D008213:Lymphocyte Activation; D051379:Mice; D011401:Promoter Regions, Genetic; D011948:Receptors, Antigen, T-Cell / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology*; D014162:Transfection
D000941:Antigens; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins