vif-Negative Human Immunodeficiency Virus Type 1 Persistently Replicates in Primary Macrophages, Producing Attenuated Progeny Virus

Authors

Iqbal Chowdhury, St. Luke’s/Roosevelt Hospital Center, College of Physicians and Surgeons, Columbia University, New York, New York
Wei Chao, St. Luke’s/Roosevelt Hospital Center, College of Physicians and Surgeons, Columbia University, New York, New York
Mary Jane Potash, St. Luke’s/Roosevelt Hospital Center, College of Physicians and Surgeons, Columbia University, New York, New York
Pavel Sova, St. Luke’s/Roosevelt Hospital Center, College of Physicians and Surgeons, Columbia University, New York, New York
Howard Gendelman, University of Nebraska Medical Center & Nebraska Center for VirologyFollow
David J. Volsky, St. Luke’s/Roosevelt Hospital Center, College of Physicians and Surgeons, Columbia University, New York, New York

Document Type

Article

Date of this Version

August 1996

Comments

Published in JOURNAL OF VIROLOGY, Aug. 1996, p. 5336–5345 Vol. 70, No. 8. 0022-538X/96/$04.0010 Copyright © 1996, American Society for Microbiology. Used by permission.

Abstract

The vif gene of human immunodeficiency virus type 1 (HIV-1) is required for efficient infection of primary T lymphocytes. In this study, we investigated in detail the role of vif in productive infection of primary monocyte-derived macrophages (MDM). Viruses carrying missense or deletion mutations in vif were constructed on the background of the monocytotropic recombinant NLHXADA-GP. Using MDM from multiple donors, we found that vif mutants produced in complementing or partially complementing cell lines were approximately 10% as infectious as wild-type virus when assayed for incomplete, complete, and circularized viral DNA molecules by quantitative PCR amplification or for viral core antigen p24 production by enzymelinked immunosorbent assay. We then determined the structure and infectivity of vif mutant HIV-1 by using MDM exclusively both for virus production and as targets for infection. Biosynthetic labeling and immunoprecipitation analysis of sucrose cushion-purified vif -negative HIV-1 made in MDM revealed that the virus had reduced p24 content compared with wild-type HIV-1. Cell-free MDM-derived vif mutant HIV-1 was infectious in macrophages as determined by the synthesis and maintenance of full-length viral DNA and by the production of particle-associated viral RNA, but its infectivity was approximately 2,500-fold lower than that of wild-type virus whose titer was determined in parallel by measurement of the viral DNA burden. MDM infected with MDM-derived vif -negative HIV-1 were able to transmit the virus to uninfected MDM by cocultivation, confirming the infectiousness of this virus. We conclude that mutations in vif significantly reduce but do not eliminate the capacity of HIV-1 to replicate and produce infectious progeny virus in primary human macrophages.