DETECTION OF BOVINE INTERLEUKIN-1-ALPHA AND INTERLEUKIN-1-BETA GENE-EXPRESSION BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION

Interleukin 1 (IL-1) is a cytokine that mediates a variety of immunoregulatory and inflammatory activities. Its role in proliferation of helper T cells is particulary important in the initiation of immune responses. Bovine interleukin 1 alpha (IL-1 alpha) and interleukin 1 beta (IL-1 beta) mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) technique in bovine peripheral blood mononuclear cells (PBMCs). Poly(A) + RNAs isolated from PBMCs by guanidium thiocyanate/oligo(dT)-cellulose chromatography extraction, were reverse-transcribed and the complementary DNAs amplified in a polymerase chain reaction primed with bovine IL-1 alpha and IL-1 beta sequence-specific primers. RT-PCR products were analyzed by agarose gel electrophoresis. As a result of the amplification of specific cDNAs by 35 cycles of PCR, the IL-1 alpha mRNA was detected from 0.01 ng of poly (A) + RNA, and IL-1 beta mRNA was detected from 0.1 pg of poly (A) + RNA isolated from PBMCs stimulated with lipopolysaccharide (LPS) for 4 h. Using 5 ng of poly(A) + RNA isolated from fresh bovine PBMCs unstimulated with any mitogen as template for RT-PCR, both IL-1 alpha and IL-1 beta mRNAs were detected at a low level.