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Journal of Virology, January 2005, p. 756-763, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.756-763.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Efficient Functional Pseudotyping of Oncoretroviral and Lentiviral Vectors by Venezuelan Equine Encephalitis Virus Envelope Proteins

Andrey A. Kolokoltsov, Scott C. Weaver, and Robert A. Davey*

Departments of Microbiology and Immunology and Pathology, University of Texas Medical Branch, Galveston, Texas

Received 7 July 2004/ Accepted 13 August 2004

Murine oncoretroviruses and lentiviruses pseudotyped with envelope proteins of alphaviruses have shown great potential in providing broad-host-range, stable vectors for gene therapy. Unlike vesicular stomatitis virus G protein-pseudotyped vectors, they are not neutralized by complement and do not appear to cause significant tissue damage. Here we report the production of murine oncoretroviral and lentiviral vectors pseudotyped with the envelope proteins of Venezuelan equine encephalitis virus (VEEV). When optimized, these pseudotypes achieve titers of 106 CFU/ml, which is 5- to 10-fold higher than for previous vectors pseudotyped with envelope proteins from other alphaviruses. They can also be concentrated or stored frozen without significant loss of infectivity. Consistent with the tropism of the envelope donor, they transduce a broad array of human cell types, including lung epithelial cells, neuronal cells, lymphocytes, and fibroblasts. Infection is blocked by agents that inhibit endosomal acidification and by neutralizing antibodies against VEEV. These observations indicate that the pseudotypes present native epitopes on their surface and enter through a VEEV envelope-dependent, pH-sensitive mechanism. The fact that the pseudotypes are unaffected by sera reactive to other alphaviruses indicates that they may be useful when successive gene therapies are required in the presence of an active immune response. In this case, having an array of alphavirus-based vectors with similar cell tropisms would be highly advantageous. These vectors may also be useful in diagnostic assays in which infectious VEEV is undesirable but immune reactivity to native epitopes is required.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Blvd., Galveston, TX 77555. Phone: (409) 772-4915. Fax: (409) 772-5065. E-mail: radavey{at}utmb.edu.

Journal of Virology, January 2005, p. 756-763, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.756-763.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Poluri, A., Ainsworth, R., Weaver, S. C., Sutton, R. E. (2008). Functional Pseudotyping of Human Immunodeficiency Virus Type 1 Vectors by Western Equine Encephalitis Virus Envelope Glycoprotein. J. Virol. 82: 12580-12584 [Abstract] [Full Text]  
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