Analysis of the function of IL-10 in chickens using specific neutralising antibodies and a sensitive capture ELISA.
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, 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-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.
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Zhiguang
Z
Wu
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Tuanjun
T
Hu
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Lisa
L
Rothwell
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Lonneke
L
Vervelde
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Pete
P
Kaiser
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Kay
K
Boulton
Department of Pathology and Pathogen Biology, Royal Veterinary College, University of London, Hatfield AL9 7TA, UK.
Matthew J
MJ
Nolan
Department of Pathology and Pathogen Biology, Royal Veterinary College, University of London, Hatfield AL9 7TA, UK.
Fiona M
FM
Tomley
Department of Pathology and Pathogen Biology, Royal Veterinary College, University of London, Hatfield AL9 7TA, UK.
Damer P
DP
Blake
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK. Electronic address: david.hume@roslin.ed.ac.uk.
David A
DA
Hume
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D057134:Antibodies, Neutralizing / Q000378:metabolism; D005075:Biological Evolution; D002478:Cells, Cultured; D002645:Chickens / Q000276:immunology*; D003048:Coccidiosis / Q000276:immunology*; D016786:Eimeria tenella / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007109:Immunity; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology*; D008322:Mammals; D051379:Mice
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D016753:Interleukin-10
Capture ELISA; Chicken; Interleukin-10; Monoclonal antibody; Neutralising antibody