Chicken-embryo fibroblasts produce two types of interferon upon stimulation with Newcastle disease virus

Abstract

Controversy has long surrounded the question of whether chickens, like mammals, can produce two types of interferon (IFN). Recently, type-I and type-II chicken IFNs have been cloned. Our study focuses on the further characterization of native fibroblast and spleen IFNs and shows that chicken embryo fibroblasts (CEFs) produce a mixture of type-I and type-II IFNs. IFN was purified by three different methods, controlled pore-glass chromatography, ion-exchange chromatography and preparative SDS-PAGE. Three protein bands showing IFN-like anti-viral activity, from CEFs which had been virus-stimulated for IFN production, were detected at 25, 27 and 29 kDa. Polyclonal antibodies produced against these bands showed partial cross-reaction with purified media from mitogen-stimulated spleen cells in ELISA, western blot analysis and anti-viral activity neutralization assay. Differences between purified conditioned media from CEF and spleen were found with respect to the stimulation of macrophages for nitric oxide production, pH stability and signal transduction pathways; only CEF IFN activated the IFN-stimulated gene factor-3 complex, whereas both CEF and spleen IFNs activated the IFN regulatory factor-1 gene. These findings concur with the differences that are known to exist between mammalian type-I and type-II IFNs. Attempts at sequencing the 25 and 27 kDa proteins by Edman degradation yielded evidence of N-terminal blockage.