DETECTION OF CYTOKINE TRANSCRIPTIONAL PROFILES FROM BOVINE PERIPHERAL-BLOOD MONONUCLEAR-CELLS AND CD4(-TRANSCRIPTASE POLYMERASE CHAIN-REACTION() LYMPHOCYTES BY REVERSE)
J. Covert et G. Splitter, DETECTION OF CYTOKINE TRANSCRIPTIONAL PROFILES FROM BOVINE PERIPHERAL-BLOOD MONONUCLEAR-CELLS AND CD4(-TRANSCRIPTASE POLYMERASE CHAIN-REACTION() LYMPHOCYTES BY REVERSE), Veterinary immunology and immunopathology, 49(1-2), 1995, pp. 39-50
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.