DEVELOPMENT OF ASSAYS TO MEASURE PORCINE IL-10 AND THEIR APPLICATION TO ANALYZING IMMUNE RESPONSES IN SWINE.

Authors: MARTIN, S - PHARMACIA AND UPJOHN; GRIMM, S - PHARMACIA AND UPJOHN; LOWERY, D - PHARMACIA AND UPJOHN; DESILVER, D - PHARMACIA AND UPJOHN; GUIMOND, P - PHARMACIA AND UPJOHN; ZUCKERMANN, F - DEPT VET UNIV ILLINOIS; Boyd, Patricia; Lunney, Joan

Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 2/13/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Many studies in murine and human systems have demonstrated that cytokines play a central role in the induction and regulation of immune responses. IFN-gamma (IFN-g) biases the immune system towards a CMI dominated Thl response, whereas IL-10 inhibits IFN-g production, downregulating Thl responses and favoring an antibody dominated Th2 response. To assess the role of cytokines in controlling porcine immune responses we initiated a joint program of cloning, expressing and purifying the individual cytokines, and then developing assays to measure their levels and enumerate the cells that produce them during immune responses. We report the cloning of porcine IL-10 by RT-PCR from activated monocytes, and its production and purification from E. Coli and baculovirus expression systems. Recombinantly produced material was used to generate two IL-10 specific monoclonal antibodies. Use of these reagents together with polyclonal sera has allowed the generation of a preliminary ELISA assay that can detect IL-10 in culture fluids in high pg/mL quantities. Work continues to refine the ELISA to increase its sensitivity, and to develop ELISPOT assays so that leukocytes producing IL-10 may be enumerated. Once these assays are in place we will use them to examine ongoing swine immune responses for IFN-g and IL-10 production. Data will be collected to examine the role of Thl or Th2 dominated responses in swine.