Journal
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
Volume 63, Issue -, Pages 206-212Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.dci.2016.04.016
Keywords
Chicken; Interleukin-10; Monoclonal antibody; Capture ELISA; Neutralising antibody
Categories
Funding
- Wellcome Trust [099164/Z/12/Z]
- BBSRC Animal Research Club [BB/L004003/1, BB/L004046/1]
- Biotechnology and Biological Sciences Research Council [BBS/E/D/20320000, BB/L004003/1, BB/L004046/1, BBS/E/D/20231760, BBS/E/D/20231759] Funding Source: researchfish
- BBSRC [BBS/E/D/20320000, BBS/E/D/20231760, BB/L004003/1, BB/L004046/1, BBS/E/D/20231759] Funding Source: UKRI
- Wellcome Trust [099164/Z/12/Z] Funding Source: Wellcome Trust
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In mammals, the inducible cytokine interleukin 10 is a feedback negative regulator of inflammation. To determine the extent to which this function is conserved in birds, recombinant chicken IL-10 was expressed as a secreted human Ig Fc fusion protein (chIL-10-Fc) and used to immunise mice. Five monoclonal antibodies (mAb) which specifically recognise chicken IL-10 were generated and characterised. Two capture ELISA assays were developed which detected native chIL-10 secreted from chicken bone marrow-derived macrophages (chBMMs) stimulated with lipppolysaccharide (LPS). Three of the mAbs detected intracellular IL-10. This was detected in only a subset of the same LPS-stimulated chBMMs. The ELISA assay also detected massive increases in circulating IL-10 in chickens challenged with the coccidial parasite, Eimeria tenella. The same mAbs neutralised the bioactivity of recombinant chIL-10. The role of IL-10 in feedback control was tested in vitro. The neutralising antibodies prevented IL-10-induced inhibition of IFN-gamma synthesis by mitogen-activated lymphocytes and increased nitric oxide production in LPS-stimulated chBMMs. The results confirm that IL-10 is an inducible feedback regulator of immune response in chickens, and could be the target for improved vaccine efficacy or breeding strategies. (C) 2016 The Authors. Published by Elsevier Ltd.
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