DETECTION OF CYTOKINE TRANSCRIPTIONAL PROFILES FROM BOVINE PERIPHERAL-BLOOD MONONUCLEAR-CELLS AND CD4(-TRANSCRIPTASE POLYMERASE CHAIN-REACTION() LYMPHOCYTES BY REVERSE)

Cytokine transcriptional profiles constitute important information abo ut T cell mediated immunity against pathogens. We have developed a rev erse transcriptase polymerase chain reaction (RT-PCR) assay to monitor gene expression of bovine T lymphocyte cytokines. Bovine cDNA was rev erse transcribed from total RNA and subsequently amplified using prime rs designed from bovine or murine and human consensus sequences. Cytok ine transcription of beta-actin, interleukins (IL) IL-1 alpha, IL-1 be ta, IL-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)alpha, TNF bet a and interferon-gamma was detected from concanavalin A activated peri pheral blood mononuclear cells and CD4(+) purified T lymphocytes. The assay allows detection of many cytokine mRNAs in a species where resea rch has been hindered by lack of commercially available reagents and s equence information. RT-PCR analysis of lymphocyte cytokines will be i nvaluable for understanding the progression or resolution of disease a s a consequence of lymphocyte response to specific antigens.