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A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
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Restricted HIV-1 replication in placental macrophages is caused by inefficient viral transcription

K García-Crespo1, C Cadilla, R Skolasky

  • 1Department of Microbiology and Medical Zoology, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico.

Journal of Leukocyte Biology
|January 1, 2010
PubMed

Insights

Human immunodeficiency virus (HIV) replication is restricted in primary monocytes (PM) compared to monocyte-derived macrophages (MDM). This restriction occurs at the transcriptional level, not viral integration.

Area of Science:

  • Virology
  • Immunology
  • Molecular Biology

Background:

  • Primary monocytes (PM) exhibit restricted human immunodeficiency virus (HIV) replication compared to monocyte-derived macrophages (MDM).
  • The precise stage of the viral replication cycle where this restriction occurs in PM remains unclear.

Purpose of the Study:

  • To investigate the specific point in the HIV replication cycle responsible for restricted replication in PM versus MDM.
  • To elucidate the molecular mechanisms underlying HIV restriction in PM.

Main Methods:

  • Infection of PM and MDM with HIV-1 BaL.
  • Alu-LTR PCR to assess proviral DNA integration.
  • Real-time RT-PCR to quantify env and gag mRNA levels.
  • Western blot analysis for viral protein expression (gp120).

Main Results:

  • No significant differences in HIV integration were observed between PM and MDM.
  • PM showed significantly lower levels of viral protein gp120 compared to MDM.
  • Real-time RT-PCR revealed substantially reduced env (24-fold less) and gag (10-fold less) mRNA levels in PM.

Conclusions:

  • HIV-1 restriction in PM occurs at the transcriptional level, impacting viral gene expression.
  • Understanding HIV restriction in PM advances knowledge of HIV pathogenesis and potential reduction in vertical transmission.

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