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Journal of Virology, August 2008, p. 7533-7539, Vol. 82, No. 15
0022-538X/08/$08.00+0     doi:10.1128/JVI.00185-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Asparagine 631 Variants of the Chicken Mx Protein Do Not Inhibit Influenza Virus Replication in Primary Chicken Embryo Fibroblasts or In Vitro Surrogate Assays{triangledown}

Camilla T. O. Benfield,1 Jon W. Lyall,1 Georg Kochs,2 and Laurence S. Tiley1*

Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom,1 Department of Virology, University of Freiburg, D-79008 Freiburg, Germany2

Received 25 January 2008/ Accepted 20 May 2008

Whether chicken Mx inhibits influenza virus replication is an important question with regard to strategies aimed at enhancing influenza resistance in domestic flocks. The Asn631 polymorphism of the chicken Mx protein found in the Shamo (SHK) chicken line was previously reported to be crucial for the antiviral activity of this highly polymorphic chicken gene. Our aims were to determine whether cells from commercial chicken lines containing Asn631 alleles were resistant to influenza virus infection and to investigate the effects that other polymorphisms might have on Mx function. Unexpectedly, we found that the Asn631 genotype had no impact on multicycle replication of influenza virus (A/WSN/33 [H1N1]) in primary chicken embryo fibroblast lines. Furthermore, expression of the Shamo (SHK) chicken Mx protein in transfected 293T cells did not inhibit viral gene expression (A/PR/8/34 [H1N1], A/Duck/England/62 [H4N6], and A/Duck/Singapore/97 [H5N3]). Lastly, in minireplicon systems (A/PR/8/34 and A/Turkey/England/50-92/91 [H5N1]), which were highly sensitive to inhibition by the murine Mx1 and human MxA proteins, respectively, Shamo chicken Mx also proved ineffective in the context of avian as well as mammalian cell backgrounds. Our findings demonstrate that Asn631 chicken Mx alleles do not inhibit influenza virus replication of the five strains tested here and efforts to increase the frequency of Asn631 alleles in commercial chicken populations are not warranted. Nevertheless, chicken Mx variants with anti-influenza activity might still exist. The flow cytometry and minireplicon assays described herein could be used as efficient functional screens to identify such active chicken Mx alleles.

* Corresponding author. Mailing address: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 OES, United Kingdom. Phone: 44 1223 339554. Fax: 44 1223 337610. E-mail: Lst21{at}cam.ac.uk

{triangledown} Published ahead of print on 28 May 2008.

Journal of Virology, August 2008, p. 7533-7539, Vol. 82, No. 15
0022-538X/08/$08.00+0     doi:10.1128/JVI.00185-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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