Development of a simple assay to measure an integral pro-inflammatory cytokine in songbird blood


Meeting Abstract

P3.194  Tuesday, Jan. 6  Development of a simple assay to measure an integral pro-inflammatory cytokine in songbird blood COON, Courtney A*; ADELMAN, Jim; LIEBL, Andrea L; MARTIN, Lynn B; University of South Florida, Tampa; Princeton University; University of South Florida, Tampa; University of South Florida, Tampa ccoon@mail.usf.edu

Progress in ecological immunology has been hampered by a lack of techniques providing strong inference into the mechanisms of disease resistance. One important integrative immune response, the acute phase response, involves adaptive alterations in body temperature, behavior and circulating proteins all which serve to control or eliminate infection. The cytokine IL-1beta is an important regulator of these proinflammatory defenses in birds. In chickens, IL-1beta, primarily secreted by macrophages, peaks in the blood 3-5 hours after injection with gram-negative bacterial components (e.g. lipopolysaccharide, LPS). Recent advent of a polyclonal antibody to chicken IL-1beta prompted us to test whether IL-1beta is detectable using the same antibody via an enzyme-linked immunosorbent assay (ELISA) in wild birds. If so, this commerically available antibody would be a useful tool in avian ecological immunology. We found that IL-1beta from wild house sparrows (Passer domesticus) was detectable 5h post-injection with complete Freunds adjuvant (CFA), a substance that mimics a bacterial infection without pathogen replication. Sparrows unchallenged with CFA showed some antibody binding, but less than CFA-treated birds. Studies are ongoing to i) validate that the antibody is binding IL-1beta and ii) develop a standard that can be used to quantify inter-assay variation.

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