|KIM, WOO - Gyeongsang National University|
|MIN, WONGI - Gyeongsang National University|
|SULLIVAN, YVONNE - Kingfisher Biotech, Inc|
|KAKACH, LAURA - Kingfisher Biotech, Inc|
|LABRESH, JOANNA - Kingfisher Biotech, Inc|
Submitted to: Developmental and Comparative Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/20/2017
Publication Date: 7/6/2017
Publication URL: http://handle.nal.usda.gov/10113/5638181
Citation: Kim, W.H., Min, W., Sullivan, Y.B., Kakach, L., Labresh, J.W., Lillehoj, H.S. 2017. Development and characterization of mouse monoclonal antibody reactive with chicken IL-8. Developmental and Comparative Immunology. 72:30-36. doi: 10.1016/j.dci.2017.02.008.
Interpretive Summary: Lack of scientific information on chicken proteins that mediate host immunity to pathogens hinders our ability to develop novel disease treatment strategy against poultry diseases. In this paper, ARS scientists collaborated with scientists in South Korea to produce recombinant chicken protein designated as CXCL8. CXCL8 plays an important role in inflammation and wound healing. Compared to similar proteins which have been identified in human and other mammals, chicken CXCL8 protein exerts a different function. To develop immune reagents which can detect chicken CXCL8 protein and its response to inflammation, ARS scientists developed mouse cell lines which secrete antibodies against chicken CXCL8 protein. These antibodies showed specific binding activity against recombinant chicken CXCL8 protein and did not recognize similar proteins from other animals. Because of their specificity to poultry immune protein which mediates important function, these immune reagents will serve as valuable immune tools for basic and applied research in poultry.
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. Since IL-8 has functions to attract lymphocytes to sites of tissue damage, it plays a role in inflammatory responses and wound healing. Chicken chemotactic and angiogenic factor (cCAF), encoded by 9E3/CEF4 gene, was identified in 1987 and has been studied as a homologue of human IL-8 for more than 20 years. However, there have not been any monoclonal antibodies (mAbs) which specifically detect chicken IL-8 (chIL-8) and the current study carried out to develop and characterize mouse mAbs against chIL-8 to further define its immunological and biological properties. Two mouse mAbs against chIL-8 were generated using recombinant chIL-8 protein produced in Escherichia coli and were confirmed by specific binding with not only recombinants but also native chIL-8 by Western blot, ELISA, and immunocytochemistry. Inhibition of IL-8-induced proliferation of chicken macrophage cells (HD11) and expression of alpha smooth muscle actin (a-SMA) in chicken embryonic fibroblast (CEF) cells by both antibodies indicated that both antibodies can be used to block bioactivity of chIL-8. These chIL-8 mAbs will be useful reagents for future investigations of inflammation in poultry.