Title |
Analysis of the function of IL-10 in chickens using specific neutralising antibodies and a sensitive capture ELISA
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Published in |
Developmental & Comparative Immunology, April 2016
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DOI | 10.1016/j.dci.2016.04.016 |
Pubmed ID | |
Authors |
Zhiguang Wu, Tuanjun Hu, Lisa Rothwell, Lonneke Vervelde, Pete Kaiser, Kay Boulton, Matthew J. Nolan, Fiona M. Tomley, Damer P. Blake, David A. Hume |
Abstract |
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 lipopolysaccharide (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, E. tenella, and the level correlated with the lesion score. 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-γ 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. |
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