Tubulovesicular Structures Within Vesicular Stomatitis Virus G Protein-pseudotyped Lentiviral Vector Preparations Carry DNA and Stimulate Antiviral Responses Via Toll-like Receptor 9

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2006 Nov 3

PMID 17079284

Citations 37

Authors

Andreas Pichlmair

Sandra S Diebold

Stephen Gschmeissner

Yasuhiro Takeuchi

Yasuhiro Ikeda

Mary K Collins

Caetano Reis e Sousa

Affiliations

Soon will be listed here.

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.

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