In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide.
In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide.
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DOI：
10.1371/journal.pone.0009310
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发表时间：
2010-02-19
期刊：
影响因子：
3.7
通讯作者：
Dezzutti CS
中科院分区：
文献类型：
--
作者：
Rohan LC;Moncla BJ;Kunjara Na Ayudhya RP;Cost M;Huang Y;Gai F;Billitto N;Lynam JD;Pryke K;Graebing P;Hopkins N;Rooney JF;Friend D;Dezzutti CS
替诺福韦凝胶已进入临床试验，用作局部杀菌剂以预防 HIV-1 感染，但尚未公布有关使用体外和离体模型进行临床前测试的数据。为了用正在进行的临床试验结果验证我们的发现，我们评估了外用替诺福韦凝胶的安全性和有效性。我们还模拟了替诺福韦的全身应用的疗效。替诺福韦凝胶的制剂评估包括渗透压、粘度、体外释放和渗透性测试。通过测量对阴道菌群、PBMC、上皮细胞以及宫颈外和结直肠外植体组织活力的影响来评估安全性。为了进行功效测试，用替诺福韦或载体对照凝胶和代表 A、B 和 C 亚型的 HIV-1 培养 PBMC。此外，用任一凝胶对极化的宫颈外植体和结直肠外植体培养物进行顶部处理。基底外侧添加替诺福韦以模拟全身应用。所有组织都受到顶部应用的 HIV-1 的攻击。通过 ELISA 测量收集的上清液中的 p24 和对宫颈外植体进行免疫组织化学分析来评估感染情况。制剂测试显示替诺福韦和赋形剂对照凝胶的等渗性>10倍。宫颈外组织的渗透性各不相同，但在所有情况下，受体区室药物浓度均达到体外抑制 HIV-1 感染的水平。该凝胶对阴道菌群、PBMC 或上皮细胞无毒。注意到宫颈外植体和结直肠外植体的上皮单层完整性和上皮断裂的短暂降低，这可能是由于制剂的高渗性质所致。替诺福韦凝胶可预防 PBMC 的 HIV-1 感染，无论 HIV-1 亚型如何。局部和全身替诺福韦可有效预防外植体培养物的 HIV-1 感染。这些研究为临床前预测配制的杀菌剂的安全性和功效提供了机制。在我们的算法中，替诺福韦对 HIV-1 感染有效。这些数据支持使用替诺福韦进行暴露前预防。
Tenofovir gel has entered into clinical trials for use as a topical microbicide to prevent HIV-1 infection but has no published data regarding pre-clinical testing using in vitro and ex vivo models. To validate our findings with on-going clinical trial results, we evaluated topical tenofovir gel for safety and efficacy. We also modeled systemic application of tenofovir for efficacy. Formulation assessment of tenofovir gel included osmolality, viscosity, in vitro release, and permeability testing. Safety was evaluated by measuring the effect on the viability of vaginal flora, PBMCs, epithelial cells, and ectocervical and colorectal explant tissues. For efficacy testing, PBMCs were cultured with tenofovir or vehicle control gels and HIV-1 representing subtypes A, B, and C. Additionally, polarized ectocervical and colorectal explant cultures were treated apically with either gel. Tenofovir was added basolaterally to simulate systemic application. All tissues were challenged with HIV-1 applied apically. Infection was assessed by measuring p24 by ELISA on collected supernatants and immunohistochemistry for ectocervical explants. Formulation testing showed the tenofovir and vehicle control gels were >10 times isosmolar. Permeability through ectocervical tissue was variable but in all cases the receptor compartment drug concentration reached levels that inhibit HIV-1 infection in vitro. The gels were non-toxic toward vaginal flora, PBMCs, or epithelial cells. A transient reduction in epithelial monolayer integrity and epithelial fracture for ectocervical and colorectal explants was noted and likely due to the hyperosmolar nature of the formulation. Tenofovir gel prevented HIV-1 infection of PBMCs regardless of HIV-1 subtype. Topical and systemic tenofovir were effective at preventing HIV-1 infection of explant cultures. These studies provide a mechanism for pre-clinical prediction of safety and efficacy of formulated microbicides. Tenofovir was effective against HIV-1 infection in our algorithm. These data support the use of tenofovir for pre-exposure prophylaxis.
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