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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #381586

Research Project: Non-antibiotic Strategies to Control Enteric Diseases of Poultry

Location: Animal Biosciences & Biotechnology Laboratory

Title: Unveiling the immunological attributes of chicken tumor necrosis factor-a (TNF-a) using new sets of monoclonal antibodies specific for poultry TNF-a

item LU, MINGMIN - US Department Of Agriculture (USDA)
item LEE, YOUNGSUB - US Department Of Agriculture (USDA)
item Lillehoj, Hyun

Submitted to: American Association of Immunologists Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 4/1/2021
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Tumor necrosis factor-a (TNF-a) is a type II transmembrane protein with either membrane-bound or soluble forms and is a prototypical member of TNF superfamily. TNF-a is a pleiotropic cytokine associated with the regulation of systemic inflammation and host defense. Chicken TNF-a (chTNF-a) is a long missed avian ortholog and the immunological properties of chTNF-a remains largely unknown compared to the mammalian counterpart. Here, we report on the functional characterization and immunomodulatory roles of chTNF-a using a panel of new anti-chTNF-a mouse monoclonal antibodies (mAbs) that we developed. A chTNF-a-specific antigen-capture ELISA was developed using compatible mAb partners via the screening and validation of ten different mAbs (3G11, 4C4, 4F3, 6A52, 6E6, 10E8, 12G6, 12H7, 14H2 and 15G7). Employing 3G11 and 12G6 as capture and detection antibodies, respectively, the levels of native chTNF-a in the circulation of Clostridium perfringens-, Eimeria- or dual C. perfringens/Eimeria-infected chickens were determined. Furthermore, intracellular expressions of chTNF-a in primary cells or cell lines derived from chickens were validated in immunocytochemistry and flow cytometry assays using both 3G11 and 12G6 mAbs. Importantly, both 3G11 and 12G6 neutralized chTNF-a-induced nitric oxide production in chicken HD11 cells in vitro. Collectively, our results provide a better understanding of the functional characteristics of chTNF-a and these anti-chTNF-a mAbs will serve as valuable immunological tools for the fundamental and applied studies in avian species. Acknowledgements: This research was supported by USDA NIFA grant on “Development of Poultry Immune Reagent” (award number: 2017-67015-26793).