Lentivirus vectors expressing short hairpin RNAs against the U3-overlapping region of HIV nef inhibit HIV replication and infectivity in primary macrophages.
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.
Yamamoto
Takuya
T
Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
Miyoshi
Hiroyuki
H
Yamamoto
Norio
N
Yamamoto
Naoki
N
Inoue
Jun-Ichiro
J
Tsunetsugu-Yokota
Yasuko
Y
D002460:Cell Line; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D015693:Gene Products, nef / Q000037:antagonists & inhibitors / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006678:HIV / Q000187:drug effects* / Q000472:pathogenicity; D015658:HIV Infections / Q000628:therapy*; D006801:Humans; D016086:Lentivirus; D019402:Macrophage Inflammatory Proteins / Q000096:biosynthesis; D008264:Macrophages / Q000821:virology; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000821:virology; D014774:Virulence / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects*; D054311:nef Gene Products, Human Immunodeficiency Virus
D054405:Chemokine CCL3; D054407:Chemokine CCL4; D015693:Gene Products, nef; D019402:Macrophage Inflammatory Proteins; D034741:RNA, Small Interfering; D054311:nef Gene Products, Human Immunodeficiency Virus