Characterisation and quantification of equine interferon gamma.
Interferon-gamma (IFN-gamma) is a key cytokine in cell-mediated immunity. To measure IFN-gamma production of equine lymphocytes (eqIFN-gamma), we developed a quantitative ELISA. Monoclonal antibodies (mAb) were produced against bacterially derived eqIFN-gamma. The mAbs recognised recombinant and lymphocyte-derived eqIFN-gamma in ELISA, Western blotting, as well as flow cytometric and microscopic analysis. In contrast to bacterially derived material, mammalian and insect cell-derived eqIFN-gamma was biologically active but could be neutralised by one of the monoclonal antibodies. Unexpectedly, glycosylation seemed to be required for antiviral activity of eqIFN-gamma.
Gutmann
Sabine
S
Deutsches Krebsforschungszentrum Heidelberg, Germany.
Zawatzky
Rainer
R
Müller
Martin
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry / Q000662:veterinary; D006736:Horses / Q000276:immunology*; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D008214:Lymphocytes / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D007371:Interferon-gamma