Detection of chicken interleukin-23, a heterodimeric cytokine with monoclonal antibodies reacting p19 and p40 subunits

Authors: KIM, WOO - US Department Of Agriculture (USDA); YEASEUL, LIM - US Department Of Agriculture (USDA); Lillehoj, Hyun

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 2/1/2018
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two covalently linked subunits, p19 (IL-23a) and p40 (IL-12/IL-23ß) which is shared with IL-12. IL-23 plays an important bridging role between the innate and adaptive immune responses. Recently, chicken IL-23 (chIL-23) has characterized and shown functional similarity to mammalian IL-23 in production of IFN-' from T cells and macrophages. In the present study, thus, we developed mouse monoclonal antibodies (mAbs) against two chIL-23 subunits, p19 and p40 and characterized their specificities in various immunoassays. To differentially detect IL-12 and IL-23 which share p40 subunit, both were expressed functionally by cotransfection with genes encoding IL-12a (also known as p35 subunit) and IL-12/IL-23ß, or IL-23a and IL-12/IL23ß into mammalian system. By several immunoassays including Western blot, and immunocytochemistry, several IL-23a mAbs react with IL-23 were found out. We also found some IL-12/23ß mAbs react with IL-12 and IL-23 and some react with IL-23 only, but not with IL-12. Using these mAbs, we developed capture ELISA assay to quantify native IL-23 from various samples. One of the mAbs also exhibited its activity to neutralize the IL-23 in production of IFN-' in chicken lymphocytes. We believe that these mAbs will be important immune reagents for basic and applied research of IL-23 in poultry.