Eszterhas Susan K, Ilonzo Nicole O, Crozier Jennifer E, Celaj Stela, Howell Alexandra L
V.A. Medical Center, White River Junction, VT; ; Department of Microbiology & Immunology, Dartmouth Medical School, Lebanon, NH, USA.
Department of Microbiology & Immunology, Dartmouth Medical School, Lebanon, NH, USA.
Infect Dis Rep. 2011 Sep 7;3(2):e11. doi: 10.4081/idr.2011.e11.
Human Immunodeficiency Virus-type 1 (HIV-1) binds to CD4 and CCR5 receptors on target cells in the human female reproductive tract. We sought to determine whether reducing levels of messenger RNA (mRNA) transcripts that encode these receptors in female reproductive tract cells could protect mucosal tissue explants from HIV-1 infection. Explants prepared from the endometrium, endocervix, and ectocervix of hysterectomy tissues from HIV-1 sero-negative women were exposed to nanoparticles containing CD4- and CCR5-specific short-interfering RNA (siRNA) sequences. Explants were then exposed two days later to HIV-1, and HIV-1 reverse transcripts were measured five days post-infection. Explants treated with nanoparticles containing CD4- and CCR5-specific siRNA showed reduced levels of CD4 and CCR5 transcripts, and significantly lower levels of HIV-1 reverse transcripts compared to those treated with an irrelevant siRNA. In female reproductive tract explants and in peripheral blood cell cultures, siRNA transfection induced the secretion of IFN-alpha (IFN-α), a potent antiviral cytokine. In female mice, murine-specific Cd4-siRNA nanoparticles instilled within the uterus significantly reduced murine Cd4 transcripts by day 3. Our findings demonstrate that siRNA nanoparticles reduce expression of HIV-1 infectivity receptors in human female reproductive tract tissues and also inhibit HIV-1 infection. Murine studies demonstrate that nanoparticles can penetrate the reproductive tract tissues in vivo and silence gene expression. The induction of IFN-α after siRNA transfection can potentially contribute to the antiviral effect. These findings support the therapeutic development of nanoparticles to deliver siRNA molecules to silence host cell receptors in the female reproductive tract as a novel microbicide to inhibit mucosal HIV-1 transmission.
人类免疫缺陷病毒1型（HIV-1）可与人类女性生殖道靶细胞上的CD4和CCR5受体结合。我们试图确定降低女性生殖道细胞中编码这些受体的信使核糖核酸（mRNA）转录本水平是否能保护黏膜组织外植体免受HIV-1感染。从HIV-1血清阴性女性子宫切除组织的子宫内膜、子宫颈管内膜和子宫颈外膜制备的外植体，被暴露于含有CD4和CCR5特异性小干扰RNA（siRNA）序列的纳米颗粒中。两天后，将外植体暴露于HIV-1，在感染后五天测量HIV-1逆转录产物。与用无关siRNA处理的外植体相比，用含有CD4和CCR5特异性siRNA的纳米颗粒处理的外植体显示CD4和CCR5转录本水平降低，且HIV-1逆转录产物水平显著降低。在女性生殖道外植体和外周血细胞培养物中，siRNA转染诱导了一种强效抗病毒细胞因子α干扰素（IFN-α）的分泌。在雌性小鼠中，子宫内注入小鼠特异性Cd4-siRNA纳米颗粒在第3天显著降低了小鼠Cd4转录本。我们的研究结果表明，siRNA纳米颗粒可降低人类女性生殖道组织中HIV-1感染性受体的表达，并抑制HIV-1感染。小鼠研究表明，纳米颗粒可在体内穿透生殖道组织并使基因表达沉默。siRNA转染后IFN-α的诱导可能有助于抗病毒作用。这些研究结果支持了纳米颗粒的治疗性开发，即将siRNA分子递送至女性生殖道以沉默宿主细胞受体，作为一种新型杀菌剂来抑制黏膜HIV-1传播。
