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Journal of Virology, June 2001, p. 5448-5456, Vol. 75, No. 12
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.12.5448-5456.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Postentry Restriction to Human Immunodeficiency Virus-Based Vector Transduction in Human Monocytes

Stuart Neil, Francisco Martin, Yasuhiro Ikeda, and Mary Collins*

Department of Immunology and Molecular Pathology, Windeyer Institute of Medical Sciences, University College London, London, United Kingdom

Received 6 December 2000/Accepted 8 March 2001

Cells of the monocyte lineage can be infected with human immunodeficiency virus type 1 (HIV-1) both during clinical infection and in vitro. The ability of HIV-1-based vectors to transduce human monocytes, monocyte-derived macrophages, and dendritic cells (DCs) was therefore examined, in order to develop an efficient protocol for antigen gene delivery to human antigen-presenting cells. Freshly isolated monocytes were refractory to HIV-1-based vector transduction but became transducible after in vitro differentiation to mature macrophages. This maturation-dependent transduction was independent of the HIV-1 accessory proteins Vif, Vpr, Vpu, and Nef in the packaging cells and of the central polypurine tract in the vector, and it was also observed with a vesicular stomatitis virus-pseudotyped HIV-1 provirus, defective only in envelope and Nef. The level and extent of reverse transcription of the HIV-1-based vector was similar after infection of immature monocytes and of mature macrophages. However, 2LTR vector circles could not be detected in monocytes, suggesting a block to vector nuclear entry in these cells. Transduction of freshly isolated monocytes exposed to HIV-1-based vector could be rescued by subsequent differentiation into DCs. This rescue was induced by fetal calf serum in the DC culture medium, which promoted vector nuclear entry.

* Corresponding author. Mailing address: Department of Immunology and Molecular Pathology, University College London, Windeyer Institute of Medical Sciences, 46 Cleveland St., London W1P 6DB, United Kingdom. Phone and fax: 44-207-679-9301. E-mail: mary.collins{at}ucl.ac.uk.

Journal of Virology, June 2001, p. 5448-5456, Vol. 75, No. 12
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.12.5448-5456.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.