Vet Med - Czech, 2021, 66(5):197-207 | DOI: 10.17221/106/2020-VETMED

Antiviral potential and stability analysis of chicken interferon-α produced by Newcastle disease virus in chicken embryo fibroblast cellsOriginal Paper

FR Anjum1, SU Rahman1, MA Aslam1, AS Qureshi2
1 Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan
2 Department of Anatomy, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan

Chicken interferon-α (chIFN-α) is an important antiviral cytokine and represents one of the first lines of the chicken's innate immune system. The current study is the first-ever report of chicken IFN (chIFN) production in Pakistan. In this study, we have used live and UV-irradiated Newcastle disease virus (NDV) to induce the expression of chIFN-α in chicken embryo fibroblast (CEF) cells. ChIFN-α was partially purified in a two-step protocol; ultracentrifugation followed by treatment with anti-chIFN-β antibodies. The purified chIFN-α was ana-lysed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the in vitro antiviral potential of chIFN-α was determined against the H9N2 avian influenza virus (AIV) via a cytopathic inhibition assay. The relative mRNA level of the IFN-stimulated genes (ISGs) in the IFN-stimulated CEF cells was measured at various time intervals by a quantitative polymerase chain reaction (qPCR). The stability of natural chIFN-α to the temperature, pH, and ultraviolet (UV) light was also determined. The in vivo therapeutic potential of chIFN-α was determined in 7-day-old broiler chickens challenged with AIV. We found that a higher chIFN-α expression level was induced by the UV-irradiated NDV in the CEF cells as compared to the live NDV. The UV-irradiated NDV induced the maximum IFN production in the CEF cells at 24 h post-infection. Two bands of 21 kDa on SDS-PAGE confirmed the presence of the chIFN-α protein. The cytopathic inhibition assay indicated the strong antiviral activity of chIFN-α against AIV. Our results of the stability analysis showed that chIFN-α was stable at a wide range of temperatures and pH levels. However, a little exposure to UV-light resulted in a significant loss of antiviral activity. We also observed that the antiviral activity of chIFN-α is related to the expression levels of the antiviral ISGs. The results of the in vivo study showed that the chIFN-α therapy via the oral route resulted in a significant improvement in the tracheal pathology of chickens challenged with AIV. In conclusion, we suggest that chIFN-α could be an important therapeutic tool to control avian influenza infection in poultry.

Keywords: antiviral; chicken embryo fibroblast; chicken type I IFNs; cytopathic inhibition assay; IFN-stimulated genes; innate immunity; Newcastle disease virus

Received: May 13, 2020; Accepted: January 6, 2021; Prepublished online: May 3, 2021; Published: May 31, 2021  Show citation

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Anjum F, Rahman S, Aslam, Qureshi A. Antiviral potential and stability analysis of chicken interferon-α produced by Newcastle disease virus in chicken embryo fibroblast cells. Vet Med - Czech. 2021;66(5):197-207. doi: 10.17221/106/2020-VETMED.
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