Abstract
Mx proteins are interferon-induced members of the dynamin superfamily of large guanosine triphosphatases. These proteins have attracted much attention because some display antiviral activity against pathogenic RNA viruses, such as members of the orthomyxoviridae, bunyaviridae, and rhabdoviridae families. Among the diverse mammalian Mx proteins examined so far, we have recently demonstrated in vitro that the Bos taurus isoform 1 (boMx1) is endowed with exceptional anti-rabies-virus activity. This finding has prompted us to seek an appropriate in vivo model for confirming and evaluating gene therapy strategies. Using a BAC transgene, we have generated transgenic mouse lines expressing the antiviral boMx1 protein and boMx2 proteins under the control of their natural promoter and short- and long-range regulatory elements. Expressed boMx1 and boMx2 are correctly assembled, as deduced from mRNA sequencing and western blotting. Poly-I/C-subordinated expression of boMx1 was detected in various organs by immunohistochemistry, and transgenic lines were readily classified as high- or low-expression lines on the basis of tissue boMx1 concentrations measured by ELISA. Poly-I/C-induced Madin-Darby bovine kidney cells, bovine turbinate cells, and cultured cells from high-expression line of transgenic mice were found to contain about the same concentration of boMx1, suggesting that this protein is produced at near-physiological levels. Furthermore, insertion of the bovine Mx system rendered transgenic mice resistant to vesicular-stomatitis-virus-associated morbidity and mortality, and embryonic fibroblasts derived from high-expression transgenic mice were far less permissive to the virus. These results demonstrate that the Bos taurus Mx system is a powerful anti-VSV agent in vivo and suggest that the transgenic mouse lines generated here constitute a good model for studying in vivo the various antiviral functions—known and yet to be discovered—exerted by bovine Mx proteins, with priority emphasis on the antirabic function of boMx1.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arnheiter H, Skuntz S, Noteborn M et al (1990) Transgenic mice with intracellular immunity to influenza virus. Cell 62:51–61
Arnheiter H, Frese M, Kambadur R et al (1996) Mx transgenic mice: animal models of health. Curr Top Microbiol Immunol 206:119–147
Babiker H, Nakatsu Y, Yamada K et al (2007) Bovine and water buffalo Mx2 genes: polymorphism and antiviral activity. Immunogenetics 59:59–67. doi:10.1007/s00251-006-0167-5
Baise E, Pire G, Leroy M et al (2004) Conditional expression of type I interferon-induced bovine Mx1 GTPase in a stable transgenic vero cell line interferes with replication of vesicular stomatitis virus. J Interferon Cytokine Res 24:513–521
Bui Tran Anh D, Faisca P, Desmecht D (2006) Differential resistance/susceptibility patterns to pneumovirus infection among inbred mouse strains. Am J Physiol Lung Cell Mol Physiol 291:L426–L435. doi:10.1152/ajplung.00483.2005
Chesler D, Reiss C (2002) IL-12, while beneficial, is not essential for the host response to VSV encephalitis. J Neuroimmunol 131:92–97. doi:10.1016/S0165-5728(02)00257-6
Faisca P, Bui Tran Anh D, Desmecht D (2005) Sendai virus-induced alterations in lung structure/function correlate with viral loads and reveal a wide resistance/susceptibility spectrum among mouse strains. Am J Physiol Lung Cell Mol Physiol 289:L777–L787. doi:10.1152/ajplung.00240.2005
Fraser R (1990) The genetics of resistance to plant viruses. Annu Rev Phytopathol 28:179–200. doi:10.1146/annurev.py.28.090190.001143
Frengen E, Weichenhan D, Zhao B et al (1999) A modular, positive selection bacterial artificial chromosome vector with multiple cloning sites. Genomics 58:250–253. doi:10.1006/geno.1998.5693
Gérardin JA, Baise EA, Pire GA et al (2004) Genomic structure, organisation, and promoter analysis of the bovine (Bos taurus) Mx1 gene. Gene 326:67–75. doi:10.1016/j.gene.2003.10.006
Giraldo P, Montoliu L (2001) Size matters: use of YACs, BACs and PACs in transgenic animals. Transgenic Res 10:83–103. doi:10.1023/A:1008918913249
Gong S, Zheng C, Doughty ML et al (2003) A gene expression atlas of the central nervous system based on bacterial artificial chromosomes. Nature 425:917–925. doi:10.1038/nature02033
Green M (1989) Catalog of mutant genes and polymorphic loci. In: Green M (ed) Genetic variants and strains of the laboratory mouse, 2nd edn. Oxford University Press, London/New York, pp 12–403
Haller O (1981) Inborn resistance of mice to orthomyxoviruses. Curr Top Microbiol Immunol 92:25–52
Haller O, Stertz S, Kochs G (2007) The Mx GTPase family of interferon-induced antiviral proteins. Microbes Infect 9:1636–1643. doi:10.1016/j.micinf.2007.09.010
Jin HK, Yamashita T, Ochiae K et al (1998) Characterization and expression of the Mx1 gene in wild mouse species. Biochem Genet 36:311–322. doi:10.1023/A:1018741312058
Kolb E, Laine E, Strehler D et al (1992) Resistance to influenza virus infection of Mx transgenic mice expressing Mx protein under the control of two constitutive promoters. J Virol 66:1709–1716
Leroy M, Baise E, Pire G et al (2005) Resistance of paramyxoviridae to type I interferon-induced Bos taurus Mx1 dynamin. J Interferon Cytokine Res 25:192–201. doi:10.1089/jir.2005.25.192
Leroy M, Pire G, Baise E et al (2006) Expression of the interferon-alpha/beta-inducible bovine Mx1 dynamin interferes with replication of rabies virus. Neurobiol Dis 21:515–521. doi:10.1016/j.nbd.2005.08.015
Lindenmann J (1962) Resistance of mice to mouse-adapted influenza A virus. Virology 16:203–204. doi:10.1016/0042-6822(62)90297-0
Müller M, Brenig B, Winnacker E et al (1992) Transgenic pigs carrying cDNA copies encoding the murine Mx1 protein which confers resistance to influenza virus infection. Gene 16:263–270. doi:10.1016/0378-1119(92)90130-H
Núñez J, Blanco E, Hernández T et al (1998) RT-PCR assay for the differential diagnosis of vesicular viral diseases of swine. J Virol Methods 72:227–235. doi:10.1016/S0166-0934(98)00032-9
Pavlovic J, Arzet H, Hefti H et al (1995) Enhanced virus resistance of transgenic mice expressing the human MxA protein. J Virol 69:4506–4510
Staeheli P, Grob R, Meier E et al (1988) Influenza virus-susceptible mice carry Mx genes with a large deletion or a nonsense mutation. Mol Cell Biol 8:4518–4523
Vanlaere I, Vanderrijst A, Guénet JL et al (2008) Mx1 causes resistance against influenza A viruses in the Mus spretus-derived inbred mouse strain SPRET/Ei. Cytokine 42:62–70. doi:10.1016/j.cyto.2008.01.013
Zhu Y, Xu G, Patel A et al (2002) Cloning, expression and initial characterization of a novel cytokine like gene family. Genomics 80:144–150. doi:10.1006/geno.2002.6816
Acknowledgments
The authors thank Dr. Fabien Ectors for his enthusiasm and exceptional skill at performing the 350 pronuclear microinjections and implanting transformed blastocysts into pseudo-pregnant mouse recipients. Many thanks are also due to Anne Cornet for genotyping the Mx1 locus of the different mouse lines.
Author information
Authors and Affiliations
Corresponding author
Additional information
M.-M. Garigliany and K. Cloquette have contributed equally to the study.
Rights and permissions
About this article
Cite this article
Garigliany, MM., Cloquette, K., Leroy, M. et al. Modulating mouse innate immunity to RNA viruses by expressing the Bos taurus Mx system. Transgenic Res 18, 719–732 (2009). https://doi.org/10.1007/s11248-009-9268-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-009-9268-x