HIV-1-based defective lentiviral vectors efficiently transduce human monocytes-derived macrophages and suppress replication of wild-type HIV-1

Lingbing Zeng, Vicente Planelles, Ziye Sui, Suzanne Gartner, Sanjay B. Maggirwar, Stephen Dewhurst, Linbai Ye, Vivek R. Nerurkar, Richard Yanagihara, Yuanan Lu

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Human monocytes play an important role in mediating human immunodeficiency virus type 1 (HIV- 1) infection of the central nervous system (CNS), and monocytes-derived macrophages (MDM) represent a major viral reservoir within the brain and other target organs. Current gene transduction of MDM is hindered by a limited efficiency. In this study we established a lentiviral vector-based technique for improved gene transfer into human MDM cultures in vitro and demonstrated significant protection of transduced MDM from super-infection with wild-type HIV-1. Methods: HIV-1-based lentiviral vector stocks were prepared in 293T cells by the established calcium phosphate transfection method. Human monocytes were isolated from donors' blood by Ficoll-Paque separation and cultured in vitro. To establish an effective technique for vector-mediated gene transfer, primary cultures of human MDM were transduced at varying multiplicities of infection (MOI) and at a range of time points following initial isolation of cells (time-in-culture). Transduced cells were then examined for transgene (green fluorescent protein (GFP)) expression by fluorescent microscopy and reverse transcription polymerase chain reaction (RT-PCR). These cultures were then exposed to wild-type HIV-1, and viral replication was quantitated by p24 assay; production of neurotoxic effector molecules by the transduced MDM was also examined, using indicator neurons. Results: We have demonstrated that primary human MDM could be efficiently transduced (>50%) with concentrated HIV-1-based defective lentiviral vectors (DLV). Furthermore, DLV-mediated gene transduction was stable, and the transduced cells exhibited no apparent difference from normal MDM in terms of their morphology, viability and neurotoxin secretion. Challenge of DLV-transduced MDM cultures with HIV-1Ba-L revealed a 4- to 5-fold reduction in viral replication, as measured by p24 antigen production. This effect was associated with the mobilization of the GFP-expressing DLV construct by the wild-type virus. Conclusions: These data demonstrate the inhibition of HIV-1 replication in primary MDM, by a DLV vector that lacks any anti-HIV-1 transgene. These findings lay the initial groundwork for future studies on the ability of DLV-modified monocytes to introduce anti-HIV-1 genes into the CNS. Lentiviral vector-mediated gene delivery to the CNS by monocytes/ macrophages is a promising, emerging strategy for treating neuro-AIDS.

Original languageEnglish (US)
Pages (from-to)18-28
Number of pages11
JournalJournal of Gene Medicine
Volume8
Issue number1
DOIs
StatePublished - Jan 2006

Keywords

  • Blood-derived monocytes (BDM)
  • Defective lentiviral vector (DLV)
  • Green fluorescent protein (GFP)
  • HIV-1
  • Monocots-derived macrophages (MDM)
  • Transduction efficiency
  • Vector mobilization (VM)

ASJC Scopus subject areas

  • Genetics

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