首页|Immunological characterization of chicken tumor necrosis factor-alpha (TNF-alpha) using new sets of monoclonal antibodies specific for poultry TNF-alpha
Immunological characterization of chicken tumor necrosis factor-alpha (TNF-alpha) using new sets of monoclonal antibodies specific for poultry TNF-alpha
扫码查看
点击上方二维码区域，可以放大扫码查看
原文链接
NSTL
Elsevier
Tumor necrosis factor-alpha (TNF-alpha) is a type II transmembrane protein with either membrane-bound or soluble forms and is a prototypical member of the TNF superfamily. TNF-alpha is a pleiotropic cytokine associated with the regulation of systemic inflammation and host defense. Chicken TNF-alpha (chTNF-alpha) is a long-missed avian ortholog, and its immunological properties remain largely unknown compared to those of its mammalian counterparts. Here, we report the functional characterization and immunomodulatory properties of chTNF-alpha using a panel of newly developed anti-chTNF-alpha mouse monoclonal antibodies (mAbs). Using anti-chTNF-alpha mAbs, we determined the tissue expression of chTNF-alpha in lymphoid and non-lymphoid organs. A chTNF-alpha-specific antigen-capture sandwich ELISA was developed using compatible mAb partners by screening and validation of ten different mAbs. Employing 3G11 and 12G6 as capture and detection antibodies, respectively, the levels of native chTNF-alpha in the circulation of Clostridium perfringens , Eimeria , or dual C. perfringens/Eimeria-infected chickens were determined. Furthermore, intracellular expression of chTNF-alpha in primary immune cells or cell lines derived from chickens was validated by immunocytochemistry and flow cytometry assays using both 3G11 and 12G6 mAbs. Notably, both 3G11 and 12G6 neutralized chTNF-alpha-induced nitric oxide production in chicken HD11 cells in vitro. Collectively, our results enhance our understanding of the functional characteristics of chTNF-alpha, and these anti-chTNF-alpha mAbs will serve as valuable immune reagents to inform on inflammatory responses and disease pathogenesis in the fundamental and applied studies of avian species.
NSTL
Elsevier
