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Journal of Virology, September 2001, p. 7925-7933, Vol. 75, No. 17
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.17.7925-7933.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Highly Productive Infection with Pseudotyped Human Immunodeficiency Virus Type 1 (HIV-1) Indicates No Intracellular Restrictions to HIV-1 Replication in Primary Human Astrocytes

Mario Canki,1 Janice Ngee Foong Thai,1 Wei Chao,1 Anuja Ghorpade,2 Mary Jane Potash,1 and David J. Volsky1,*

Division of Molecular Virology, St. Luke's-Roosevelt Hospital Center and Columbia University, New York, New York 10019,1 and Center for Neurovirology and Neurodegenerative Disorders, University of Nebraska Medical Center, Omana, Nebraska 681982

Received 12 March 2001/Accepted 24 May 2001

Human astrocytes can be infected with human immunodeficiency virus type 1 (HIV-1) in vitro and in vivo, but, in contrast to T lymphocytes and macrophages, virus expression is inefficient. To investigate the HIV-1 life cycle in human fetal astrocytes, we infected cells with HIV-1 pseudotyped with envelope glycoproteins of either amphotropic murine leukemia virus or vesicular stomatitis virus. Infection by both pseudotypes was productive and long lasting and reached a peak of 68% infected cells and 1.7 µg of viral p24 per ml of culture supernatant 7 days after virus inoculation and then continued with gradually declining levels of virus expression through 7 weeks of follow-up. This contrasted with less than 0.1% HIV-1 antigen-positive cells and 400 pg of extracellular p24 per ml at the peak of astrocyte infection with native HIV-1. Cell viability and growth kinetics were similar in infected and control cells. Northern blot analysis revealed the presence of major HIV-1 RNA species of 9, 4, and 2 kb in astrocytes exposed to pseudotyped (but not wild-type) HIV-1 at 2, 14, and 28 days after infection. Consistent with productive infection, the 9- and 4-kb viral transcripts in astrocytes infected by pseudotyped HIV-1 were as abundant as the 2-kb mRNA during 4 weeks of follow-up, and both structural and regulatory viral proteins were detected in infected cells by immunoblotting or cell staining. The progeny virus released by these cells was infectious. These results indicate that the major barrier to HIV-1 infection of primary astrocytes is at virus entry and that astrocytes have no intrinsic intracellular restriction to efficient HIV-1 replication.

* Corresponding author. Mailing address: Division of Molecular Virology, St. Luke's-Roosevelt Hospital Center, 432 West 58th St., Antenucci Research Bldg., Rm. 709, New York, NY 10019. Phone: (212) 582-4451. Fax: (212) 582-5027. E-mail: djv4{at}columbia.edu.

Journal of Virology, September 2001, p. 7925-7933, Vol. 75, No. 17
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.17.7925-7933.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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