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Journal of Virology, March 1999, p. 1828-1834, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Requirements for Efficient Production and Transduction of Human Immunodeficiency Virus Type 1-Based Vectors

Mehdi Gasmi, Jacqueline Glynn, Ming-Jie Jin, Douglas J. Jolly, Jiing-Kuan Yee, and Shin-Tai Chen*

Center for Gene Therapy, Chiron Technologies, San Diego, California 92121

Received 28 August 1998/Accepted 12 November 1998

A number of human immunodeficiency type 1 (HIV-1)-based vectors have recently been shown to transduce nondividing cells in vivo as well as in vitro. However, if these vectors are to be considered for eventual clinical use, a major consideration is to reduce the probability of unintended generation of replication-competent virus. This can be achieved by eliminating viral genetic elements involved in the generation of replication-competent virus without impairing vector production. We have designed a system to transiently produce HIV-1-based vectors by using expression plasmids encoding Gag, Pol, and Tat of HIV-1 under the control of the cytomegalovirus immediate-early promoter. Our data show that the best vector yield is achieved in the presence of the Rev/Rev-responsive element (RRE) system. However, the constitutive transport element of Mason-Pfizer monkey virus can substitute for RRE and Rev at least to some extent, whereas the posttranscriptional regulatory element of human hepatitis B virus appeared to be inefficient. In addition, we show that high-titer virus preparations can be obtained in the presence of sodium butyrate, which activates the expression of both the packaging construct and the vector genome. Finally, our results suggest that efficient infectivity of vectors defective in the accessory proteins Vif, Vpr, Vpu, and Nef depends on the nature of the target cells.

* Corresponding author. Present address: Jerry L. Pettis VAMC and Loma Linda University, Mineral Metabolism (151), 11201 Benton St., Loma Linda, CA 92357. Phone: (909) 422-3101. Fax: (909) 796-1680. E-mail: STChen{at}netscape.net.

Journal of Virology, March 1999, p. 1828-1834, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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