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Journal of Virology, April 2001, p. 3371-3382, Vol. 75, No. 7
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.7.3371-3382.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Construction and Molecular Analysis of Gene Transfer Systems Derived from Bovine Immunodeficiency Virus

Robert Berkowitz,* Heini Ilves, Wei Yu Lin, Karl Eckert, Andrea Coward, Stan Tamaki, Gabor Veres, and Ivan Plavec

Systemix Inc., Palo Alto, California 94304

Received 20 October 2000/Accepted 4 January 2001

Because lentiviruses are able to infect nondividing cells, these viruses might be utilized in gene therapy applications where the target cell does not divide. However, it has been suggested that the introduction of primate lentivirus sequences, particularly those of human immunodeficiency virus, into human cells may pose a health risk for the patient. To avoid this concern, we have constructed gene transfer systems based on a nonprimate lentivirus, bovine immunodeficiency virus. A panel of vectors and packaging constructs was generated and analyzed in a transient expression system for virion production and maturation, vector expression and encapsidation, and envelope protein pseudotyping. Virion preparations were also analyzed for transduction efficiency in a panel of human and nonhuman primary cells and immortalized cell lines. The virion preparations transduced most of the target cell types, with efficiencies up to 90% and with titers of unconcentrated virus up to 5 × 105 infectious doses/ml. In addition, infection of nondividing human cells, including unstimulated hematopoietic stem cells and irradiated endothelial cells, was observed.

* Corresponding author. Present address: Chemocentryx, 1539 Industrial Rd., San Carlos, CA 94070. Phone: (650) 632-2900. E-mail: rberkowitz{at}chemocentryx.com.

Journal of Virology, April 2001, p. 3371-3382, Vol. 75, No. 7
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.7.3371-3382.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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