Author
KIM, WOO - US Department Of Agriculture (USDA) | |
Lillehoj, Hyun | |
JEONG, MISUN - US Department Of Agriculture (USDA) | |
MIN, WONGI - Gyeongsang National University | |
SULIVAN, YVONNE - Kingfisher Biotech, Inc | |
KAKACH, LINDA - Kingfisher Biotech, Inc | |
LABRESH, JOANNA - Kingfisher Biotech, Inc |
Submitted to: American Association for Immunology
Publication Type: Abstract Only Publication Acceptance Date: 12/3/2015 Publication Date: N/A Citation: N/A Interpretive Summary: Technical Abstract: 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. Given that IL-8 attracts lymphocytes to the sites of tissue damage, IL-8 plays a role in the inflammatory response and wound healing. Previously, chicken chemotactic and angiogenic factor which is encoded by the 9E3/CEF4 gene, was identified and has been studied as a human IL-8 homologue. However, detecting and quantifying this bioactive chemokine/cytokine in avian species is limited by the lack of specific antibodies and reliable bioassays. Thus, we developed and characterized mouse monoclonal antibodies (mAbs) against chicken IL-8 to further define its immunological and biological properties. Two mouse mAbs against chicken IL-8 were generated using recombinant IL-8 protein produced in Escherichia coli. The specificities of these antibodies were confirmed by their specific binding with not only recombinant IL-8, but also native IL-8 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 these mAbs indicate that both antibodies are capable of blocking IL-8 bioactivity. Therefore, this study demonstrates that chicken immune reagents, which detect IL-8, will be valuable for basic as well as applied studies in poultry. |