Development of antigen-capture ELISA to analyze the immunological functions of chicken interferon-kappa

Authors: LEE, YOUNGSUB - US Department Of Agriculture (USDA); LU, MINGMIN - US Department Of Agriculture (USDA); Lillehoj, Hyun

Submitted to: American Association of Avian Pathologists
Publication Type: Abstract Only
Publication Acceptance Date: 1/8/2021
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Interferon (IFN)-' is a type I IFN and plays a central role in host antiviral defense and immune response. The function of type I IFNs in birds have not been clearly defined, and limited information of IFN-' has been reported in poultry compared to its counterpart in mammals. In this study, we developed an antigen-capture ELISA that can determine the production of chicken using antigen-specific mouse monoclonal antibodies (mAbs). Recombinant chicken IFN-' expressed in E. coli was used to immunize the mice. Five mAbs that specifically recognize chicken IFN-' were selected and characterized based on their specificity and binding activity toward chicken CCL5 by indirect ELISA and western blot. For sandwich ELISA development for chicken IFN-', two sets of best capture and detection mAb combination were identified by pairing assay. IFN-' production induced by polyinosinic: polycytidylic acid (poly I: C) in chicken HD11 macrophage cells was effectively detected by the newly developed capture ELISA, and the gene expression of IFN-' was validated using qRT-PCR. Neutralizing effects of anti-chIFN-' mAbs were tested by their ability to block the induction of IFN-stimulated genes (ISGs) in DF-1 cells upon the stimulation by recombinant chIFN-' protein. Two mAbs blocked the mRNA expression of ISGs in a dose-dependent manner. The newly developed capture ELISA and anti-chicken IFN-' mAbs will serve as valuable immune reagents for poultry scientists.