School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China.
1] School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China [2] Department of Pharmacy, Guangdong Provincial Stomatological Hospital, Guangzhou, China.
Lab Invest. 2014 Jul;94(7):777-87. doi: 10.1038/labinvest.2014.72. Epub 2014 May 19.
The breakdown of human retinal pigment epithelial (HRPE) barrier is considered as the etiology of retinopathy, which affects the quality of life of HIV/AIDS patients. Here we demonstrate that HIV-1 could directly impair HRPE barrier function, which leads to the translocation of HIV-1 and bacteria. HRPE cells (D407) were grown to form polarized, confluent monolayers and treated with different HIV-1 infectious clones. A significant increase of monolayer permeability, as measured by trans-epithelial electrical resistance (TEER) and apical-basolateral movements of sodium fluorescein, was observed. Disrupted tightness of HRPE barrier was associated with the downregulation of several tight junction proteins in D407 cells, including ZO-1, Occludin, Claudin-1, Claudin-2, Claudin-3, Claudin-4, and Claudin-5, after exposure to HIV-1, without affecting the viability of cells. HIV-1 gp120 was shown to participate in the alteration of barrier properties, as evidenced by decreased TEER and weakened expression of tight junction proteins in D407 monolayers after exposure to pseudotyped HIV-1, UV-inactivated HIV-1, and free gp120, but not to an envelope (Env)-defective mutant of HIV. Furthermore, exposure to HIV-1 particles could induce the release of pro-inflammatory cytokines in D407, including IL-6 and MCP-1, both of which downregulated the expression of ZO-1 in the HRPE barrier. Disrupted HRPE monolayer allowed translocation of HIV-1 and bacteria across the epithelium. Overall, these findings suggest that HIV-1 may exploit its Env glycoprotein to induce an inflammatory state in HRPE cells, which could result in impairment of HRPE monolayer integrity, allowing virus and bacteria existing in ocular fluids to cross the epithelium and penetrate the HRPE barrier. Our study highlights the role of HIV-1 in the pathogenesis of HIV/AIDS-related retinopathy and suggests potential therapeutic targets for this ocular complication.
人视网膜色素上皮 (HRPE) 屏障的破坏被认为是发生视网膜病变的病因,这会影响 HIV/AIDS 患者的生活质量。在这里,我们证明 HIV-1 可以直接损害 HRPE 屏障功能,导致 HIV-1 和细菌的易位。将 HRPE 细胞 (D407) 培养成形成极化、融合的单层,并使用不同的 HIV-1 感染性克隆进行处理。通过跨上皮电阻 (TEER) 和钠荧光素的顶端-基底侧运动测量,观察到单层通透性显著增加。在暴露于 HIV-1 后,D407 细胞中几种紧密连接蛋白(包括 ZO-1、Occludin、Claudin-1、Claudin-2、Claudin-3、Claudin-4 和 Claudin-5)的下调与 HRPE 屏障的紧密性破坏有关,而不会影响细胞活力。已经表明 HIV-1 gp120 参与了屏障特性的改变,这可以通过在暴露于假型 HIV-1、UV 失活的 HIV-1 和游离 gp120 后 D407 单层的 TEER 降低和紧密连接蛋白表达减弱来证明,但对于 HIV 的包膜 (Env) 缺陷突变体则不是如此。此外,暴露于 HIV-1 颗粒可诱导 D407 中促炎细胞因子的释放,包括 IL-6 和 MCP-1,两者均下调 HRPE 屏障中 ZO-1 的表达。HRPE 单层的破坏允许 HIV-1 和细菌穿过上皮细胞。总的来说,这些发现表明 HIV-1 可能利用其 Env 糖蛋白在 HRPE 细胞中诱导炎症状态,这可能导致 HRPE 单层完整性受损,允许存在于眼液中的病毒和细菌穿过上皮细胞并穿透 HRPE 屏障。我们的研究强调了 HIV-1 在 HIV/AIDS 相关视网膜病变发病机制中的作用,并为这种眼部并发症提出了潜在的治疗靶点。