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The substitution U475 → C with Sabin3-like mutation within the IRES attenuate Coxsackievirus B3 cardiovirulence

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Abstract

The Sabin3 mutation in the viral RNA plays an important role in directing attenuation phenotype of Sabin vaccine strain of poliovirus type 1 (PV1). We previously described that Sabin3-like mutation introduced in Coxsackievirus B3 (CVB3) genome led to a defective mutant. However, this mutation do not led to destruction of secondary structure motif C within the stem-loop V of CVB3 RNA because of the presence of one nucleotide difference (C → U) in the region encompassing the Sabin3 mutation at nucleotides 471 of PV1 and 475 of CVB3 RNA. In order to reproduce the same sequence of PV1 sabin3 vaccine strain, we introduce in this study an additional mutation (U475 → C) to CVB3 Sabin3-like mutant. Our results demonstrated that Sabin3-like+C mutant displayed a decreased translation initiation defects when translated in cell-free system. This translation initiation defect was correlated with reduced yields of infectious virus particles in HeLa cells in comparison with Sabin3-like mutant and wild-type CVB3 viruses. Inoculation of Swiss mice with mutant viruses resulted in no inflammatory heart disease when compared to heart of mice infected with wild-type. Theses findings indicate that the double mutant could be exploited for the development of a live attenuated vaccine against CVB3.

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Acknowledgements

We thank Dr. Abdelfattah Zakhama from the college of Medicine of Monastir (Tunisia) for helpful discussion of histology data; Moncef Belayouni and Houda Daami for expert technical assistance. This work was financed by grants to J. Gharbi from the franco-tunisian cooperation: DGRST-CNRS (code 04/R0902) and CMCU (code 04/G0810). M. Ben M’hadheb-Gharbi is supported by CNRS, CMCU and MDT-01 training grants.

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  1. Unité de Pathogenèse et Virulence Virales (PVV), Laboratoire des Maladies Transmissibles (MDT-01), Université de Monastir, Faculté de Pharmacie, Avenue Avicenne, Monastir, 5000, Tunisia

    Manel Ben M’hadheb-Gharbi, Mahjoub Aouni & Jawhar Gharbi

  2. Unité de la Régulation de la Traduction Eucaryote et Virale, CNRS URA 1966, Institut Pasteur de Paris, 28 Rue du Dr. Roux, Paris, 75724, France

    Manel Ben M’hadheb-Gharbi, Sylvie Paulous & Katherine M. Kean

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  1. Manel Ben M’hadheb-Gharbi
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  2. Sylvie Paulous
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Correspondence to Jawhar Gharbi.

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M’hadheb-Gharbi, M.B., Paulous, S., Aouni, M. et al. The substitution U475 → C with Sabin3-like mutation within the IRES attenuate Coxsackievirus B3 cardiovirulence. Mol Biotechnol 36, 52–60 (2007). https://doi.org/10.1007/s12033-007-0019-z

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