Analysis of the function of IL-10 in chickens using specific neutralising antibodies and a sensitive capture ELISA

Zhiguang Wu, Tuanjun Hu, Lisa Rothwell, Lonneke Vervelde, Pete Kaiser, Kay Boulton, Matthew J Nolan, Fiona M Tomley, Damer P Blake, David A Hume

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)206-212
JournalDevelopmental and Comparative Immunology
Volume63
Early online date22 Apr 2016
DOIs
Publication statusPublished - Oct 2016

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