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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Ruminant Diseases and Immunology Research » Research » Publications at this Location » Publication #141767
Title: INTRACELLULAR BOVINE CYTOKINE ANALYSIS WITH ANTI-HUMAN CYTOKINE MABS

Author
item Kimura, Kayoko
item Goff, Jesse
item STABEL, JUDITH - 3625-30-10

Submitted to: Great Lakes International Imaging and Flow Cytometry Association
Publication Type: Abstract Only
Publication Acceptance Date: 10/5/2002
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Flow cytometric analysis of intracellular cytokines has proven useful for assay of human and rodent immune cell function. Limited availability of commercial mAbs to bovine cytokines has prevented application of this method to bovine immune cells. We have used anti-human IFN-gamma and IL-8 mAbs to develop a method to study intracellular cytokines in bovine immune cells. PBMC expressing IFN-gamma and PMN expressing IL-8 increased by 11 and 86%, respectively, following culturing with pokeweed mitogen for 20 hr. Ab to human IL-8 is known to cross-react with bovine IL-8. To validate the specificity of anti-human IFN-gamma mAb against bovine IFN-gamma, we incubated the mAb with recombinant bovine IFN-gamma and demonstrated nearly complete blockage of mAb binding to cells. These results demonstrate anti-human cytokine mAbs can be used for intracellular bovine cytokine analysis. In the course of these studies we also demonstrated bovine neutrophils are positive for IFN-gamma production as reported in human cells.