Enhancing dengue virus maturation using a stable furin over-expressing cell line

Swati Mukherjee, Devika Sirohi, Kimberly A. Dowd, Zhenguo Chen, Michael S. Diamond, Richard J. Kuhn, Theodore C. Pierson

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Flaviviruses are positive-stranded RNA viruses that incorporate envelope (E) and premembrane (prM) proteins into the virion. Furin-mediated cleavage of prM defines a required maturation step in the flavivirus lifecycle. Inefficient prM cleavage results in structurally heterogeneous virions with unique antigenic and functional characteristics. Recent studies with dengue virus suggest that viruses produced in tissue culture cells are less mature than those produced in primary cells. In this study, we describe a Vero cell line that ectopically expresses high levels of human furin (Vero-furin) for use in the production of more homogenous mature flavivirus populations. Laboratory-adapted and clinical dengue virus isolates grow efficiently in Vero-furin cells. Biochemical and structural techniques demonstrate efficient prM cleavage in Vero-furin derived virus preparations. These virions also were less sensitive to neutralization by antibodies that bind efficiently to immature virions. This furin-expressing cell line will be of considerable utility for flavivirus neutralization and structural studies.

Original languageEnglish (US)
Pages (from-to)33-40
Number of pages8
JournalVirology
Volume497
DOIs
StatePublished - Oct 1 2016

Keywords

  • Antibody neutralization
  • Dengue virus
  • Flavivirus
  • Furin
  • Virion maturation
  • West Nile virus

ASJC Scopus subject areas

  • Virology

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    Mukherjee, S., Sirohi, D., Dowd, K. A., Chen, Z., Diamond, M. S., Kuhn, R. J., & Pierson, T. C. (2016). Enhancing dengue virus maturation using a stable furin over-expressing cell line. Virology, 497, 33-40. https://doi.org/10.1016/j.virol.2016.06.022