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Journal of Virology, February 2005, p. 1765-1771, Vol. 79, No. 3
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.3.1765-1771.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Human Immunodeficiency Virus Type 1 Vectors with Alphavirus Envelope Glycoproteins Produced from Stable Packaging Cells

Blair L. Strang,1,{dagger} Yasuhiro Takeuchi,1 Thomas Relander,2 Johan Richter,2 Ranbir Bailey,1 David A. Sanders,3 Mary K. L. Collins,1 and Yasuhiro Ikeda1*

Department of Immunology and Molecular Pathology, University College London, University of London, London, United Kingdom,1 Department of Molecular Medicine and Gene Therapy, University of Lund, Lund, Sweden,2 Department of Biological Sciences, Purdue University, West Lafayette, Indiana3

Received 26 May 2004/ Accepted 7 September 2004

Alphavirus glycoproteins have broad host ranges. Human immunodeficiency virus type 1 (HIV-1) vectors pseudotyped with their glycoproteins could extend the range of tissues that can be transduced in both humans and animal models. Here, we established stable producer cell lines for HIV vectors pseudotyped with alphavirus Ross River virus (RRV) and Semliki Forest virus (SFV) glycoproteins E2E1. RRV E2E1-stable clones could routinely produce high-titer pseudotyped vectors for at least 5 months. SFV E2E1-stable clones, however, produced relatively low titers. We examined the properties of RRV E2E1-pseudotyped vectors [HIV-1(RRV)] and compared them with amphotropic murine leukemia virus Env- and vesicular stomatitis virus glycoprotein G-pseudotyped vectors. HIV-1(RRV) displayed a number of characteristics which would be advantageous in ex vivo and in vivo experiments, including resistance to inactivation by heat-labile components in fresh human sera and thermostability at 37°C. Upon single-step concentration by ultracentrifugation of HIV-1(RRV), we could achieve vector stocks with titers up to 6 x 107 IU/ml. HIV-1(RRV) efficiently transduced cells from several different species, including murine primary dendritic cells, but failed to transduce human and murine T cells as well as human hematopoietic stem cells (HSC). These results indicate that HIV-1(RRV) could be used in a number of applications including animal model experiments and suggest that expression of RRV cellular receptors is limited or absent in certain cell types such as T cells and human HSC.

* Corresponding author. Present address: Molecular Medicine Program, Mayo Clinic College of Medicine, 200 First St. SW, Rochester, MN 55905. Phone: (507) 538-0153. Fax: (507) 266-2122. E-mail: Ikeda.Yasuhiro{at}mayo.edu.

{dagger} Present address: MRC Virology Unit, Institute of Virology, Glasgow, United Kingdom.

Journal of Virology, February 2005, p. 1765-1771, Vol. 79, No. 3
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.3.1765-1771.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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