Tubulovesicular structures within Vesicular stomatitis virus G protein-pseudotyped lentiviral vector preparations carry DNA and stimulate antiviral responses via toll-like receptor 9

Andreas Pichlmair, Sandra S. Diebold, Stephen Gschmeissner, Yasuhiro Takeuchi, Yasuhiro H Ikeda, Mary K. Collins, Caetano Reis E Sousa

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Recombinant lentiviral vectors (LVs) are commonly used as research tools and are being tested in the clinic as delivery agents for gene therapy. Here, we show that Vesicular stomatitis virus G protein (VSV-G)-pseudotyped LV preparations produced by transient transfection are heavily contaminated with tubulovesicular structures (TVS) of cellular origin, which carry nucleic acids, including the DNA plasmids originally used for LV generation. The DNA carried by TVS can act as a stimulus for innate antiviral responses, triggering Toll-like receptor 9 and inducing alpha/beta interferon production by plasmacytoid dendritic cells (pDC). Removal of TVS markedly reduces the ability of VSV-G-pseudotyped LV preparations to activate pDC. Conversely, virus-free TVS are sufficient to stimulate pDC and act as potent adjuvants in vivo, eliciting T- and B-cell responses to coadministered proteins. These results highlight the role of by-products of virus production in determining the immunostimulatory properties of recombinant virus preparations and suggest possible strategies for diminishing responses to LVs in gene therapy and in research use.

Original languageEnglish (US)
Pages (from-to)539-547
Number of pages9
JournalJournal of Virology
Volume81
Issue number2
DOIs
StatePublished - Jan 2007

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Toll-Like Receptor 9
Vesiculovirus
dendritic cells
G-proteins
Dendritic Cells
Antiviral Agents
gene therapy
Viruses
Genetic Therapy
viruses
DNA
interferon-beta
Interferon-beta
cell structures
Cellular Structures
transfection
Research
Interferon-alpha
Nucleic Acids
nucleic acids

ASJC Scopus subject areas

  • Immunology

Cite this

Tubulovesicular structures within Vesicular stomatitis virus G protein-pseudotyped lentiviral vector preparations carry DNA and stimulate antiviral responses via toll-like receptor 9. / Pichlmair, Andreas; Diebold, Sandra S.; Gschmeissner, Stephen; Takeuchi, Yasuhiro; Ikeda, Yasuhiro H; Collins, Mary K.; Reis E Sousa, Caetano.

In: Journal of Virology, Vol. 81, No. 2, 01.2007, p. 539-547.

Research output: Contribution to journalArticle

Pichlmair, Andreas ; Diebold, Sandra S. ; Gschmeissner, Stephen ; Takeuchi, Yasuhiro ; Ikeda, Yasuhiro H ; Collins, Mary K. ; Reis E Sousa, Caetano. / Tubulovesicular structures within Vesicular stomatitis virus G protein-pseudotyped lentiviral vector preparations carry DNA and stimulate antiviral responses via toll-like receptor 9. In: Journal of Virology. 2007 ; Vol. 81, No. 2. pp. 539-547.
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