Lentivirus vectors expressing short hairpin RNAs against the U3-overlapping region of HIV nef inhibit HIV replication and infectivity in primary macrophages

Blood. 2006 Nov 15;108(10):3305-12. doi: 10.1182/blood-2006-04-014829. Epub 2006 Jul 20.


Although successful attempts to inhibit HIV-1 replication in T cells using RNAi have been reported, the effect of HIV-specific RNAi on macrophages is not well known. Macrophages are key targets for anti-HIV-1 therapy because they are able to survive long after the initial infection with HIV and can spread the virus to T cells. In this study, we identified a putative RNAi target of HIV, consisting of the portion of the nef gene overlapping the U3 region (Nef366), and generated a lentivirus-based short hairpin RNA (shRNA) expression vector (Lenti shNef366). We show that Lenti shNef366 inhibits (1) HIV-1 replication in a monocytic cell line and in primary monocyte-derived macrophages (MDMs), (2) reactivation of latent HIV-1 infection, and (3) the production of secondary HIV-1 from MDMs harboring a genomic copy of Nef366. Moreover, we found that the up-regulated production of macrophage inflammatory protein 1beta (MIP-1beta), but not MIP-1alpha, in MDMs by Nef expression was considerably suppressed by Lenti shNef366, which suggests that HIV-1 dissemination to T cells through its interaction with HIV-1-infected MDMs can also be controlled by Lenti shNef366. Thus, lentivirus-mediated shRNA expression targeting the U3-overlapping region of HIV nef represents a feasible approach to genetic vaccine therapy for HIV-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Chemokine CCL3
  • Chemokine CCL4
  • Gene Products, nef / antagonists & inhibitors
  • Gene Products, nef / genetics*
  • Genetic Therapy / methods
  • Genetic Vectors
  • HIV / drug effects*
  • HIV / pathogenicity
  • HIV Infections / therapy*
  • Humans
  • Lentivirus
  • Macrophage Inflammatory Proteins / biosynthesis
  • Macrophages / virology
  • RNA, Small Interfering / administration & dosage*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology*
  • T-Lymphocytes / virology
  • Virulence / drug effects
  • Virus Replication / drug effects*
  • nef Gene Products, Human Immunodeficiency Virus


  • Chemokine CCL3
  • Chemokine CCL4
  • Gene Products, nef
  • Macrophage Inflammatory Proteins
  • RNA, Small Interfering
  • nef Gene Products, Human Immunodeficiency Virus