A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine

Yujiao Yang, Chao Shan, Jing Zou, Antonio E. Muruato, Diniz Nunes Bruno, Barbosa de Almeida Medeiros Daniele, Pedro F C Vasconcelos, Shannan Rossi, Scott Weaver, Xuping Xie, Pei-Yong Shi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A purified inactivated vaccine (PIV) using the Zika virus (ZIKV) Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E) was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production) for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains. The NS1 mutation did not affect viral RNA synthesis, but significantly increased virion assembly through an increased interaction between NS1 and NS2A (a known regulator of flavivirus assembly). The NS1 mutant virus retained wild-type virulence in the A129 mouse model, but decreased its competence to infect Aedes aegypti mosquitoes. To further increase virus yield, we constructed an infectious cDNA clone of the clinical trial PIV strain PRVABC59 containing three viral replication-enhancing mutations (NS1 K265E, prM H83R, and NS3 S356F). The mutant cDNA clone produced >. 25-fold more ZIKV than the wild-type parent on Vero cells. This cDNA clone-launched manufacture platform has the advantages of higher virus yield, shortened manufacture time, and minimized chance of contamination.

Original languageEnglish (US)
JournalEBioMedicine
DOIs
StateAccepted/In press - Jan 4 2017

Fingerprint

Inactivated Vaccines
Viruses
Complementary DNA
Clone Cells
Puerto Rico
Vero Cells
Vaccines
Flavivirus
Clinical Trials, Phase I
Mutation
Aedes
Viral RNA
Virus Replication
Culicidae
Virion
Mental Competency
Haplorhini
Virulence
Clinical Trials
Cell Line

Keywords

  • Flavivirus assembly
  • Flavivirus NS1
  • Zika virus vaccine

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Yang, Y., Shan, C., Zou, J., Muruato, A. E., Bruno, D. N., de Almeida Medeiros Daniele, B., ... Shi, P-Y. (Accepted/In press). A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine. EBioMedicine. https://doi.org/10.1016/j.ebiom.2017.02.003

A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine. / Yang, Yujiao; Shan, Chao; Zou, Jing; Muruato, Antonio E.; Bruno, Diniz Nunes; de Almeida Medeiros Daniele, Barbosa; Vasconcelos, Pedro F C; Rossi, Shannan; Weaver, Scott; Xie, Xuping; Shi, Pei-Yong.

In: EBioMedicine, 04.01.2017.

Research output: Contribution to journalArticle

Yang, Y, Shan, C, Zou, J, Muruato, AE, Bruno, DN, de Almeida Medeiros Daniele, B, Vasconcelos, PFC, Rossi, S, Weaver, S, Xie, X & Shi, P-Y 2017, 'A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine', EBioMedicine. https://doi.org/10.1016/j.ebiom.2017.02.003
Yang Y, Shan C, Zou J, Muruato AE, Bruno DN, de Almeida Medeiros Daniele B et al. A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine. EBioMedicine. 2017 Jan 4. https://doi.org/10.1016/j.ebiom.2017.02.003
Yang, Yujiao ; Shan, Chao ; Zou, Jing ; Muruato, Antonio E. ; Bruno, Diniz Nunes ; de Almeida Medeiros Daniele, Barbosa ; Vasconcelos, Pedro F C ; Rossi, Shannan ; Weaver, Scott ; Xie, Xuping ; Shi, Pei-Yong. / A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine. In: EBioMedicine. 2017.
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