Detection of chicken interleukin-23, a heterodimeric cytokine with monoclonal antibodies reacting p19 and p40 subunits
| Autor: | Woohyun Kim, Yeaseul Lim, Hyun S. Lillehoj |
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| Rok vydání: | 2018 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 200:109.10-109.10 |
| ISSN: | 1550-6606 0022-1767 |
| Popis: | Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two covalently linked subunits, p19 (IL-23α) and p40 (IL-12/IL-23β) 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-γ 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-12α (also known as p35 subunit) and IL-12/IL-23β, or IL-23α and IL-12/IL23β into mammalian system. By several immunoassays including Western blot, and immunocytochemistry, several IL-23α mAbs react with IL-23 were found out. We also found some IL-12/23β 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-γ in chicken lymphocytes. We believe that these mAbs will be important immune reagents for basic and applied research of IL-23 in poultry. |
| Databáze: | OpenAIRE |
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