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

Abstract

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.

In Vitro Characterization of chIFITMs of Aseel and Kadaknath Chicken Breeds against Newcastle Disease Virus Infection

“…It is also involved in inhibiting cell adhesion and controlling cell growth [46]. Additionally, Anjum et al noticed a decrease in cytopathic effects in chIFN-treated CEF cells when they were infected with ND and AIV [47]. Similarly, in Kadaknath, CEF cells expressed a high level of IFN-γ,and delayed cytopathic changes were observed.…”

Production of an ultrasound-assisted biosurfactant postbiotic from agro-industrial wastes and its activity against Newcastle virus

“…The strain was supplied from school of veterinary medicine, Shiraz university, Shiraz, Iran. A mixure of different doses of biosurfactants (100 μL) and virus strains (100 μL) supplemented with antibiotic and antifungal agents were propagated into three 9-day-old chicken embryo eggs and then incubated in a humidified condition at 37°C for 48 h. The eggs were moved to refrigerator at 4°C and then the allantoic fluids were collected and the presence and activity of the virus strains in the allantoic floids of each eggs were examined using haemagglutination (HA) assay ( 34 – 36 ).…”

Role of interferons in the antiviral battle: from virus-host crosstalk to prophylactic and therapeutic potential in SARS-CoV-2 infection

“…Human IFNs are proteins of 130-170 amino acids with a molecular weight of 20 - 100 kDa, active at temperatures generally lower than 56-60°C and at different pH values ( 47 – 51 ). IFNα and IFNβ (are acid-stable, while IFNγ is acid-labile ( 13 , 52 ).…”