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United States Department of Agriculture

Agricultural Research Service

Title: Measurement of Bovine Interleukin-12 Mrna Expression by the Reverse Transcriptase-Polymerase Chain Reaction

item Stabel, Judith
item Kehrli Jr, Marcus
item Stevens, Mark - FORMER USDA, ARS, NADC

Submitted to: International Virtual Conference on Infectious Diseases of Animals
Publication Type: Abstract Only
Publication Acceptance Date: April 20, 1997
Publication Date: N/A

Technical Abstract: A reverse transcriptase-polymerase chain (RT-PCR) was developed to amplify and clone the 35 and 40 kDa subunits of bovine interleukin-12 (IL-12) using bovine IL-12-specific primers. The RT-PCR detected IL-12 mRNA expression for the 35 or 45 kDa subunits, or both subunits, when bovine neutrophils from a normal calf or a calf with leukocyte adhesion deficiency were stimulated for 6 or 18 hours with Escherichia coli 055:B5 lipopolysaccharide (LPS), LPS plus human interferon-gamma (IFN-gamma), or killed Brucella abortus strain RB51. IL-12 mRNA was generally not present following stimulation of neutrophils for 24 or 48 hours. During 6 or 48 hours of incubation, unstimulated neutrophils did not produce IL-12 mRNA. IL-12 mRNA for the 35 kDa subunit was also detected by the RT-PCR in a simian virus 40 transformed bovine macrophage cell line (MAC cells) following stimulation of the cells for either 24 hours with LPS plus bovine IFN-gamma or for 6 hours with LPS plus human IFN-gamma. In addition, a bovine CD4+ T cell clone, 300B1 cells, produced mRNA for the 40 kDa IL-12 subunit when cells were stimulated for 24 hours with concanavalin A. Unstimulated MAC cells and 300B1 cells did not produce IL-12 mRNA.

Last Modified: 4/22/2015
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