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Establishment of a reverse genetics system for rotavirus vaccine strain LLR and developing vaccine candidates carrying VP7 gene cloned from human strains circulating in China
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  • Xiafei Liu,
  • Shan Li,
  • Junjie Yu,
  • Pengdi Chai,
  • Lili Pang,
  • Jin-song Li,
  • Wuyang Zhu,
  • Weihong Ren,
  • Zhaojun Duan
Xiafei Liu
The First Affiliated Hospital of Henan University of Traditional Chinese Medicine
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Shan Li
National Institute for Viral Disease Control and Prevention
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Junjie Yu
National Institute for Viral Disease Control and Prevention
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Pengdi Chai
National Institute for Viral Disease Control and Prevention
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Lili Pang
National Institute for Viral Disease Control and Prevention
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Jin-song Li
National Institute for Viral Disease Control and Prevention
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Wuyang Zhu
National Institute for Viral Disease Control and Prevention
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Weihong Ren
The First Affiliated Hospital of Henan University of Traditional Chinese Medicine
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Zhaojun Duan
National Institute for Viral Disease Control and Prevention

Corresponding Author:[email protected]

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Abstract

Human rotavirus A (RVA) causes acute gastroenteritis in infants and young children. The LLR RVA vaccine, which licensed in 2000 and widely used in China, significantly reduced rotavirus disease burden in China. With the exchanges of RV circulating strains and the emergence of new genotypes, the LLR vaccine against RVGE needed to be upgraded. In this study, we aimed to establish an RG system for the RVA vaccine strain LLR (G10P[15]). Transfection with plasmids expressing 11 genomic RNA segments of LLR along with the pCMV/868CP helper plasmid, resulted in rescue of the infectious virus with an artificially introduced genetic marker on its genome, indicating that an RG system for the LLR strain was successfully established. Furthermore, the plasmid‐based reverse genetics system was used to generate lamb RVA reassortants with VP4 and VP7 genes derived from human RVA strains in China, which were not previously adapted to cell culture. We were able to rescue the six VP7 (G1, G2, G3, G4, G8, and G9) mono‐reassortants, but no VP4 (P[4] or P[8]) mono‐reassortant was rescued. The six VP7 reassortants covered all G-genotypes currently circulating in China and stably replicated in MA104 cells, which should be exploited as the next generation rotavirus vaccines candidates in China. Furthermore, the LLR RG system in this study will be a useful vaccine vector for intestinal pathogens such as norovirus and Vibrio cholerae.
12 Jul 2024Submitted to Journal of Medical Virology
12 Jul 2024Submission Checks Completed
12 Jul 2024Assigned to Editor
12 Jul 2024Review(s) Completed, Editorial Evaluation Pending
13 Jul 2024Reviewer(s) Assigned
06 Sep 2024Editorial Decision: Revise Major
18 Oct 20241st Revision Received
25 Oct 2024Submission Checks Completed
25 Oct 2024Assigned to Editor
25 Oct 2024Review(s) Completed, Editorial Evaluation Pending
28 Oct 2024Reviewer(s) Assigned
31 Oct 2024Editorial Decision: Accept