Lentivirus vectors expressing short hairpin RNAs against the U3-overlapping region of HIV nef inhibit HIV replication and infectivity in primary macrophages
Lentivirus vectors expressing short hairpin RNAs against the U3-overlapping region of HIV nef inhibit HIV replication and infectivity in primary macrophages
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DOI：
10.1182/blood-2006-04-014829
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发表时间：
2006-11-15
期刊：
影响因子：
20.3
通讯作者：
Tsunetsugu-Yokota, Yasuko
中科院分区：
文献类型：
--
作者：
Yamamoto, Takuya;Miyoshi, Hiroyuki;Tsunetsugu-Yokota, Yasuko
尽管已经报道了使用RNAi成功地尝试抑制HIV-1在T细胞中的复制，但HIV特异性RNAi对巨噬细胞的作用尚不清楚。巨噬细胞是抗HIV-1治疗的关键靶点，因为它们能够在最初感染HIV后存活很长时间，并能将病毒传播给T细胞。在这项研究中，我们确定了一个假定的RNAi靶点，由nef基因与U3区重叠的部分组成（Nef 366），并产生了一个基于慢病毒的短发夹RNA（shRNA）表达载体（Lenti shNef 366）。我们展示了这个大斋节！shNef 366抑制（1）单核细胞系和原代单核细胞衍生的巨噬细胞（MDM）中的HIV-1复制，（2）潜伏HIV-1感染的再激活，和（3）从携带Nef 366基因组拷贝的MDM产生继发HIV-1。此外，我们发现，通过Nef表达在MDM中上调巨噬细胞炎性蛋白10（MIP-1 β）而不是MIP-1 α的产生被Lenti shNef 366显著抑制，这表明HIV-1通过与HIV-1感染的MDM相互作用传播到T细胞也可以被Lenti shNef 366控制。因此，针对HIV nef的U3重叠区域的慢病毒介导的shRNA表达代表了HIV-1基因疫苗治疗的可行方法。
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 lenti-virus-based short hairpin RNA (shRNA) expression vector (Lenti shNef366). We show that Lent! 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 10 (MIP-1 beta), but not MIP-1 alpha, 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.