Kinetics of interleukin 2 mRNA and protein produced in the human T-cell line Jurkat and effect of cyclosporin A.
The kinetics of interleukin 2 mRNA accumulation in the leukemic T-cell line Jurkat, which can be induced with phytohemagglutinin and phorbol 12-myristate 13-acetate to produce large amounts of interleukin 2, was analyzed by a modified DNA-excess solution hybridization assay using a 5'-32P-labeled oligodeoxyribonucleotide 30 bases long as probe. Cyclosporin A was used as a valuable tool to gain more insight into the quantitative aspects of interleukin 2 production, on the basis of the assumption that transcription of the interleukin 2 gene is completely inhibited shortly after administration of cyclosporin A. The half-life of interleukin 2 mRNA was estimated to be approximately 2 h. With the aid of simple mathematical models, we have been able to relate the concentration of interleukin 2 protein in the supernatant to the interleukin 2 mRNA kinetics. This novel quantitative kinetic analysis revealed that, independent of the absence or presence of cyclosporin A, interleukin 2 protein is synthesized at a rate of approximately 1.3 molecules per molecule of interleukin 2 mRNA per second and secreted within 2 h after it is synthesized and that its half-life in the supernatant is approximately 10 h.
Nordmann
R
R
Department of Preclinical Research, Sandoz Ltd., Basle, Switzerland.
Andersen
E
E
Trussardi
R
R
Mazer
N A
NA
D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003524:Cyclosporins / Q000494:pharmacology*; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics*; D007700:Kinetics; D008433:Mathematics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology; D014158:Transcription, Genetic / Q000187:drug effects
D003524:Cyclosporins; D007376:Interleukin-2; D015345:Oligonucleotide Probes; D012333:RNA, Messenger