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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 #359775

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

Location: Animal Biosciences & Biotechnology Laboratory

Title: Development of novel monoclonal antibodies against chicken interleukin- 4 and alternative activation of macrophage-like cells in chickens

item CHAUDHARI, ATUL - US Department Of Agriculture (USDA)
item KIM, WOOHYUN - US Department Of Agriculture (USDA)
item 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: The functions of a Th2 cytokine such as interleukin 4 (IL-4) in chickens is not well understood due to the lack of specific immune reagents against chicken IL-4 (chIL-4). In mammals, IL-4 stimulate alternative activation of macrophages. However, this phenomenon has never been reported in chickens till date. In the present study, we developed mouse monoclonal antibodies (mAbs) against chIL-4 and studied alternative activation of macrophage. Alternative activation of HD11 cells was investigated by measuring nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) expression, arginase activity and gene expressions of iNOS, CD80 and CD86 (M1 markers) and chemokine (C-C motif) ligand 17 (ccl17) and mannose receptor C-type1 (MRC1L-A) (M2 markers) in HD11 cells treated with chIL-4, lipopolysaccharide (LPS), chIL-4+LPS, chIL-4+LPS+mAbs. The mAbs successfully detected the endogenously produced chIL-4 (chicken sera and stimulated HD-11 cell supernatant and pellet) by capture ELISA, ICC and flow cytometry. Further, NO production by LPS-stimulated HD11 cells and primary monocyte/macrophage cells was inhibited by chIL-4 with reduced iNOS expression and increased arginase activity. Also, chIL-4 induced robust expression of M2 marker genes than M1-related genes. All these effects were neutralized by anti-chIL-4 antibodies. In conclusion, our results showed that newly developed anti-chIL-4 mAbs could serve as valuable immune reagents to detect chIL-4 and to explore chIL-4 functionality. Additionally, our results demonstrated that chIL-4 may regulate alternative activation of HD11 cells in chicken and indicate the M1/M2 paradigm in chickens.