Comparative investigation of in terleukin-2 and in terleukin-2 receptor alpha mRNA expression in lymphocytes activated by pha or cona

One of the classic methods of lymphocytes functional activity research is blast-transformation reaction (RBTL) were proliferation rate in response to different stimuli in vitro is estimated by introduction of radioactive 3H.thymidine in cell DNA. Nonspecific mytogens phytohemagglutinin (PHA) or concanavalin A (ConA) inducing generally T-cell response are usually used as a control in RBTL. At present there are only descrete and segmentary data about expression patterns of key genes involved in lymphocytes reaction to nonspecific mitogens. The aim of our work was to obtain the integral picture of interleukin-2 (IL-2) and interleukin-2 receptor alpha (IL-2ra) mRNA expression pattern in lymphocytes during long period of cultivation after their stimulation with PHA or ConA as far as IL-2 and IL-2ra are one of the key genes involved in mononuclear cells response to nonspecific mitogen stimulation. mRNA expression of these genes was calculated by realtime PCR method that is highly sensitive and specific and allows to work in wide range of mRNA concentration. Common patterns of IL-2 and IL-2ra mRNA expression in cells of healthy donors stimulated with T-cells mitogens were identified. The obtained results allow to conclude that mechanisms of action of PHA and ConA on cells are different and that significant individual differences of mRNA kinetics in lymphocytes of distinct donors are possible. IL-2 mRNA level can be used as a marker of lymphocytes activation by ConA in the very first hours of stimulation, but with PHA stimulation the cytokine's mRNA content achieves its maximum only on 3.4 days of cultivation.