Cholesterol supplementation during production increases the infectivity of retroviral and lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G)

Yong Chen, Christopher J. Ott, Kay Townsend, Papasani Subbaiah, Ashok Aiyar, William M. Miller*

*Corresponding author for this work

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Cholesterol, a major component of plasma membrane lipid rafts, is important for assembly and budding of enveloped viruses, including influenza and HIV-1. Cholesterol depletion impairs virus assembly and infectivity. This study examined the effects of exogenous cholesterol addition (delivered as a complex with methyl-beta-cyclodextrin (MbCD)) on the production of Molony murine leukemia virus (MoMuLV) retroviral vector and HIV-1-based lentiviral vector pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G). Cholesterol supplementation before and during vector production enhanced the infectivity of retroviral and lentiviral vectors up to 4-fold and 6-fold, respectively. In contrast, the amount of retroviral vector produced was unchanged, and that of lentiviral vector was increased less than 2-fold. Both free cholesterol and cholesterol ester content in 293-gag-pol producer cells increased with cholesterol addition. In contrast, the phospholipids headgroup composition was essentially unchanged by cholesterol supplementation in 293-gag-pol packaging cells. Based on these results, it is proposed that cholesterol supplementation increases the infectivity of VSV-G-pseudotyped retroviral and lentiviral vectors, possibly by altering the composition of the producer cell membrane where the viral vectors are assembled and bud, and/or by changing the lipid composition of the viral vectors.

Original languageEnglish (US)
Pages (from-to)199-207
Number of pages9
JournalBiochemical Engineering Journal
Volume44
Issue number2-3
DOIs
StatePublished - May 15 2009

Keywords

  • Cholesterol
  • Infectivity
  • Lentiviral vector
  • Lipid rafts
  • Retroviral vector
  • Viral vector production

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Engineering
  • Bioengineering
  • Biomedical Engineering

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