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Characterization of street rabies virus variants with an additional N-glycan at position 247 in the glycoprotein

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Abstract

Most street rabies virus glycoproteins (G proteins) possess two N-glycosylation sites, at Asn37 and Asn319, whereas an additional N-glycosylation site is present in several fixed (laboratory-adapted) rabies virus strains at Asn247, which suggests that the N-glycosylation addition may be a marker of fixed viruses. In this study, we successfully cloned two street virus strain 1088 variants, N5B#15 and N5B#10-28, in which the G proteins had an additional N-glycan at position 247, and we examined whether these variants were characterized by cell culture adaptation and attenuation after intramuscular inoculation as fixed viruses. N5B#15 had four mutations, i.e., S148P and D247N in the G protein, and T137A and N2046S in the large (L) protein. N5B#10-28 had an additional mutation in the G protein, R196I. Compared with the parental 1088 virus, both variants exhibited highly efficient replication in mouse neuroblastoma-derived NA cells and reduced pathogenicity in adult mice when inoculated intramuscularly, but not intracerebrally. However, this attenuation was not attributable to the induction of strong immune responses.

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Acknowledgements

We thank Drs. Makoto Sugiyama and Naoto Ito (Gifu University, Japan) for providing the recovery system for RC-HLΔG/GFP (pRC-HLΔG/GFP, pCI-RG, pT7IRES-RN, pT7IRES-RP, pT7IRES-RL and BHK/T7-9 cells) and the anti-G MAb 15-13. This work was supported by the Ministry of Education, Culture, Sports, Science, and Technology of Japan (grant number: 21780278) and the Ministry of Health, Labour and Welfare of Japan (Health and Labour Sciences Research Grants, Research on International Cooperation in Medical Science).

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The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Research Promotion Institute, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu, Oita, 879-5593, Japan

    Kentaro Yamada, Kazuko Noguchi & Akira Nishizono

  2. Department of Microbiology, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu, Oita, 879-5593, Japan

    Kazuko Noguchi & Akira Nishizono

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  1. Kentaro Yamada
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Correspondence to Kentaro Yamada.

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Yamada, K., Noguchi, K. & Nishizono, A. Characterization of street rabies virus variants with an additional N-glycan at position 247 in the glycoprotein. Arch Virol 159, 207–216 (2014). https://doi.org/10.1007/s00705-013-1805-5

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  • DOI: https://doi.org/10.1007/s00705-013-1805-5

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