Detection of cytokine transcriptional profiles from bovine peripheral blood mononuclear cells and CD4+ lymphocytes by reverse transcriptase polymerase chain reaction.

Cytokine transcriptional profiles constitute important information about T cell mediated immunity against pathogens. We have developed a reverse transcriptase polymerase chain reaction (RT-PCR) assay to monitor gene expression of bovine T lymphocyte cytokines. Bovine cDNA was reverse transcribed from total RNA and subsequently amplified using primers designed from bovine or murine and human consensus sequences. Cytokine transcription of beta-actin, interleukins (IL) IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)alpha, TNF beta and interferon-gamma was detected from concanavalin A activated peripheral blood mononuclear cells and CD4+ purified T lymphocytes. The assay allows detection of many cytokine mRNA in a species where research has been hindered by lack of commercially available reagents and sequence information. RT-PCR analysis of lymphocyte cytokines will be invaluable for understanding the progression or resolution of disease as a consequence of lymphocyte response to specific antigens.