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Changes in the NS1 gene of avian influenza viruses isolated in Thailand affect expression of type I interferon in primary chicken embryonic fibroblast cells

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Abstract

The non-structural protein 1 (NS1) of avian influenza virus was defined as one of the virulent factors. To understand the effect of NS1 protein of influenza virus H5N1 isolated in Thailand on type I (α/β) interferon (IFN) synthesis, five reverse genetic viruses were constructed and used as models. The viruses were generated using NS genomic segment from A/Peurto Rico/8/1934 (H1N1) and four avian influenza viruses isolated from the first outbreak in Thailand. All the viruses have the rest of the genome from A/Peurto Rico/8/1934 (H1N1). The constructed viruses were named (1) NS1 PR8/34, (2) NS1 wild type, (3) NS1 L15FD53G, (4) NS1 N171I and (5) NS1 E71K, respectively. The type I (α/β) IFN gene expression in control and infected primary chicken embryonic fibroblast cells were analyzed by quantitative polymerase chain reaction. The results show that the inhibition of IFN-β gene expression by NS1 wild type infected cells is stronger than NS1 N171I, NS1 E71K, NS1 PR8/34 and NS1 L15FD53G, respectively. The data suggest that the difference of amino acid sequence of NS1 protein contributes to the IFN-β antagonist. In contrast, the difference of the NS1 protein does not influence in the IFN-α antagonistic activity.

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Acknowledgments

Faculty of Medical science and Faculty of Medicine, Naresuan University, Thailand provided laboratory instruments. Veterinary Research and Development Centre of Northern Lower Region, Phitsanulok, Thailand provided the viral specimen and biosafety level 3 facilities. The pHW2000 plasmids containing the cDNA of each gene of the A/Puerto Rico/8/34 (H1N1) were given from Dr. Robert G. Webster and Dr. Erich Hoffmann of St. Jude Children Research Hospital, Memphis, USA. We would like to thank Prof. Dr. Gavin Reynolds and Dr. Ian Fraser for his suggestion of the manuscript.

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Correspondence to Donruedee Sanguansermsri.

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Thepmalee, C., Sanguansermsri, P., Suwanankhon, N. et al. Changes in the NS1 gene of avian influenza viruses isolated in Thailand affect expression of type I interferon in primary chicken embryonic fibroblast cells. Indian J. Virol. 24, 365–372 (2013). https://doi.org/10.1007/s13337-013-0158-8

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