مرکز منطقه ای اطلاع رساني علوم و فناوري -

چكيده لاتين :

Cytokines secreted by Th1 (T-helper)/Th2 cells play an important role in the pathogenesis of many diseases. Th1 cells secrete predominantly IFN-م and IL-2 which regulate cell-mediated immunity against intracellular pathogens and tumors. In this study, expression of IFN-م was studied using semiquantitative real time polymerase chain reaction (RT-PCR). In brief, lymphocytes of a healthy donor were stimulated with PHA (1ىg/106 cell/ml) in cell culture at different incubation times (0, 4, 8, 12, 24, 48 and 72 hours) to express IFN-م. Total RNA was extracted and cDNA synthesized. A sequence (273 bp) between two oligonucleotide primers (chosen from two different exons of the IFN-م gene sequences) was amplified using a heat-stable DNA polymerase. In semi-quantitative RT-PCR, we used a serial dilution for cDNA in order to determine the titer of cDNA which gives visible band in agarose gel (2%) electrophoresis. Results showed that the highest level of IFN-م expression was achieved after 4 hours activation with PHA and it was stable at least for 22 hours. Then it fell to baseline level. Cytokine detection using RT-PCR will provide useful clinical information.