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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Research » Publications at this Location » Publication #323537

Title: Development and characterization of monoclonal antibodies specific for chicken IL-8

item KIM, WOO - US Department Of Agriculture (USDA)
item Lillehoj, Hyun
item JEONG, MISUN - US Department Of Agriculture (USDA)
item SULIVAN, YVONNE - Kingfisher Biotech, Inc
item KAKACH, LAURA - Kingfisher Biotech, Inc
item LABRESH, JOANNA - Kingfisher Biotech, Inc

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 12/3/2015
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Limited information on chicken cytokines and chemokines hinders progress in understaidng the role of cell-mediated immunity in infections. Interleukin-8/CXCL8 (IL-8) is a CXC-family chemokine produced by fibroblasts and other cell types, including epithelial cells, endothelial cells, neutrophils, and macrophages. IL-8 attracts lymphocytes to sites of tissue damage, and plays a role in the inflammatory response and wound healing. The chicken chemotactic and angiogenic factor, encoded by the 9E3/CEF4 gene, was identified and has been studied as a human IL-8 homologue for over more than 20 years. However, the detecting and quantifying of the bioactive avian specific IL-8 chemokine/cytokine is limited by the lack of specific antibodies and reliable bioassays although the use of monoclonal antibodies (mAbs) has become routine in the mammalian research and diagnosis. Thus, we developed and characterized mouse mAbs against chIL-8 to further define its immunological and biological properties. Two mouse mAbs against IL-8 were generated using recombinant chicken IL-8 protein produced in Escherichia coli. These mAbs were confirmed to show specific binding with not only recombinant IL-8, but also the native IL-8 protein by Western blot analysis, enzyme-linked immunosorbent assay, and immunocytochemistry. Inhibition of IL-8-induced chemotaxis of peripheral blood lymphocytes, proliferation of chicken macrophage cells and expression of alpha smooth-muscle actin in chicken embryonic fibroblast cells by both antibodies indicate that both antibodies are capable of blocking chIL-8 bioactivity. These results show that two mAbs have been generated that detect native IL-8 for basic immunology studies in poultry.