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.
Covert
J
J
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
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D000818:Animals; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002417:Cattle / Q000235:genetics* / Q000276:immunology*; D003208:Concanavalin A / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D016207:Cytokines; D017931:DNA Primers; D012333:RNA, Messenger; D003208:Concanavalin A