Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 18, 2011
Publication Date: June 23, 2011
Citation: Hong, Y., Lillehoj, H.S., Lee, S.H., Park, S., Min, W., Labresh, J., Tompkins, D., Baldwin, C. 2011. Development and characterization of mouse monoclonal antibodies specific for chicken interleukin 18. Veterinary Immunology and Immunopathology. 138(1-2):144-148.
Interpretive Summary: The lack of reagents which can be used to assess host immune response in poultry hinders the progress in basic and applied research for this animal species. In this paper, ARS scientist, in collaboration with scientists from other universities, describe the development of new mouse antibodies detecting a chicken cytokine 18. Chicken IL-18 gene was cloned and its protein was expressed in a yeast expression vector to test the biological function of chicken IL-18 and to make mouse monoclonal antibodies against chicken IL-18. This study showed that mouse antibodies identified recombinant and native chicken IL-18 molecules and they were able to neutralize the biological function of IL-18. This is the first paper which describes the production of mouse monoclonal antibodies detecting chicken IL-18. These antibodies will be commercialized to the scientific community for basic and applied research in poultry science.
Four mouse monoclonal antibodies (mAbs) which are specific for chicken interleukin 18 (chIL18) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting, quantitative real-time PCR and neutralization assays. Monoclonal antibodies specific for chIL18 identified a 23 kDa Pichia pastoris-expressed chIL18 and 66 kDa E. coli-derived MBP fusion protein of chIL18 by Western blot analysis. Bioassays for chIL18 using primary chicken spleen cells showed dose-dependent IFN-' mRNA expression and induction of IFN-' from primary splenocytes, and triggered nitric oxide (NO) production in HD11 chicken macrophage cell line. These mAbs showed neutralizing activity against chIL18. Taken together, these anti-chIL18 mouse mAbs will be important new immune reagents useful for basic and applied research in poultry.