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Association of Mx1 Asn631 variant alleles with reductions in morbidity, early mortality, viral shedding, and cytokine responses in chickens infected with a highly pathogenic avian influenza virus

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Abstract

Myxovirus-resistance (Mx) proteins are produced by host cells in response to type I interferons, and some members of the Mx gene family in mammals have been shown to limit replication of influenza and other viruses. According to an early report, chicken Mx1 variants encoding Asn at position 631 have antiviral activity, whereas variants with Ser at 631 lack activity in experiments evaluating Mx1 complementary DNA (cDNA) expressed ectopically in a cell line. We evaluated whether the Mx1 631 dimorphism influenced pathogenesis of highly pathogenic avian influenza virus (HPAIV) infection in chickens of two commercial broiler lines, each segregating for Asn631 and Ser631 variants. Following intranasal infection with HPAIV strain A/Chicken/Queretaro/14588-19/1995 H5N2, chickens homozygous for Asn631 allele were significantly more resistant to disease based on early mortality, morbidity, or virus shedding than Ser631 homozygotes. Higher amounts of splenic cytokine transcripts were observed in the Ser631 birds after infection, consistent with higher viral loads seen in this group and perhaps contributing to their higher morbidity. Nucleotide sequence determination of Mx1 cDNAs demonstrated that the Asn631 variants in the two chicken lines differed at several amino acid positions outside 631. In vitro experiments with a different influenza strain (low pathogenicity) failed to demonstrate an effect of Mx1 Asn631 on viral replication suggesting that in vivo responses may differ markedly from in vitro, or that choice of virus strain may be critical in demonstrating effects of chicken Mx1. Overall, these studies provide the first evidence that Mx1 has antiviral effects in chickens infected with influenza virus.

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Acknowledgments

This project was supported by the National Research Initiative Competitive Grant no. 2006-35205-16676 from the USDA National Institute of Food and Agriculture, and USDA Agricultural Research Service CRIS project no. 6612-32000-053. We thank Clayton Floyd and Roger Brock for animal care. We also thank Dr. Igal Pevzner and Mr. Randy Shumate of Cobb-Vantress, Inc., for the chickens, and Dr. Peter Staeheli (University of Freiburg, Freiburg, Germany) for the expression construct for recombinant chicken IFN-α. We gratefully acknowledge the technical expertise of Ms. Milla Kaltenboeck, Ms. Tracy Faulkner, Mr. Cam Greene, and Ms. Aniko Zsak.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Pathobiology, Auburn University, 166 Greene Hall, Auburn, AL, 36849-5519, USA

    Sandra J. Ewald

  2. Department of Poultry Science, Auburn University, Auburn, AL, USA

    Sandra J. Ewald & Emily J. Livant

  3. Southeast Poultry Research Laboratory, ARS-USDA, Athens, GA, USA

    Darrell R. Kapczynski & David L. Suarez

  4. Aviagen, Crossville, TN, USA

    John Ralph, Scott McLeod & Carolyn Miller

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  1. Sandra J. Ewald
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  2. Darrell R. Kapczynski
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  6. Scott McLeod
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Correspondence to Sandra J. Ewald.

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Ewald, S.J., Kapczynski, D.R., Livant, E.J. et al. Association of Mx1 Asn631 variant alleles with reductions in morbidity, early mortality, viral shedding, and cytokine responses in chickens infected with a highly pathogenic avian influenza virus. Immunogenetics 63, 363–375 (2011). https://doi.org/10.1007/s00251-010-0509-1

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