Development of sandwich ELISA for differential detection of chicken heterodimeric cytokines, interleukin-12 and -23

Authors: KIM, WOOHYUN - US Department Of Agriculture (USDA); Lillehoj, Hyun

Submitted to: American Association of Avian Pathologist
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2019
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Interleukin (IL) -12 cytokine family consists of IL-12, IL-23, IL-27 and IL-35. Unlike mammals, there are only two cytokines have been identified in chickens so far among the IL-12 family, IL-12 and IL-23. They are heterodimeric cytokines consisting of two covalently linked subunits, IL-12; p35 (IL-12a) and p40 (IL-12/IL-23ß) and IL-23; p19 (IL-23a) and p40. Since p40 subunit is shared between IL-12 and IL-23, it is difficult to identify the nature of cytokine if PCR-based detection method using p40 is used to measure IL-12 and IL-23 production. In the present study, we developed mouse monoclonal antibodies (mAbs) which are specific against three subunits, p19, p35 and p40, and developed the sandwich ELISA to differentiate IL-12 and IL-23 expressions. The p40 mAb was used as capture antibody and p35 or p19 mAb was used as detection antibody for IL-12 and IL-23 differentiation, respectively. Some of the new mABs we developed also neutralized the function of IL-12 and IL-23 and blocked production of IFN-' and IL-17A in chicken T cells. In summary, we have generated novel sensitive detection system to differentially identify chicken IL-12 and IL-23 at the level of protein expression. This method will be valuable to obtain an insight to understand chicken cell-mediated immunity.