Dezzutti Charlene S, Park Seo Young, Marks Kenneth M, Lawlor Sidney E, Russo Julie R, Macio Ingrid, Chappell Catherine A, Bunge Katherine E
1 Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh , Pittsburgh, Pennsylvania.
2 Magee-Womens Research Institute , Pittsburgh, Pennsylvania.
AIDS Res Hum Retroviruses. 2018 Feb;34(2):185-192. doi: 10.1089/AID.2017.0107. Epub 2017 Dec 14.
In clinical trials evaluating HIV-1 prevention products, ex vivo exposure of mucosal tissue to HIV-1 is performed to inform drug levels needed to suppress viral infection. Understanding assay and participant variables that influence HIV-1 replication will help with assay implementation. Demographic and behavioral data were obtained from 61 healthy women aged 21-45. Paired cervical tissue (CT) and vaginal tissue (VT) biopsies were collected and treated with HIV-1 or HIV-1, washed, and cultured. On days 3, 7, and/or 11, culture supernatant was collected, and viral replication was monitored by p24 ELISA. Tissue was extracted at study end, and HIV-1 relative RNA copies were determined by polymerase chain reaction. Cumulative p24 and RNA were log-transformed and analyzed using a linear mixed model, t-test, and an intraclass correlation coefficient (ICC). HIV replication was similar between CT and VT for each virus, but HIV-1 had 1.5 log and 0.9 log higher levels of p24 than HIV-1 in CT and VT, respectively (p < .001), which correlated with HIV-1 relative RNA copies. Cumulative p24 and RNA copies in both tissues demonstrated low intraperson correlation for both viruses (ICC ≤0.513 HIV-1; ICC ≤0.419 HIV-1). Enrollment into previous clinical studies in which genital biopsies were collected modestly decreased the HIV-1 cumulative p24 for CT, but not for VT. To improve the ex vivo challenge assay, viruses should be evaluated for replication in mucosal tissue before study implementation, baseline mucosal tissue is not needed if a placebo/no treatment group is included within the clinical trial, and previous biopsy sites should be avoided.
在评估HIV-1预防产品的临床试验中，进行粘膜组织对HIV-1的体外暴露以确定抑制病毒感染所需的药物水平。了解影响HIV-1复制的检测方法和参与者变量将有助于检测方法的实施。从61名年龄在21-45岁的健康女性中获取人口统计学和行为数据。采集配对的宫颈组织（CT）和阴道组织（VT）活检样本，用HIV-1或HIV-1处理，冲洗后培养。在第3天、第7天和/或第11天，收集培养上清液，通过p24 ELISA监测病毒复制。在研究结束时提取组织，通过聚合酶链反应测定HIV-1相对RNA拷贝数。对累积的p24和RNA进行对数转换，并使用线性混合模型、t检验和组内相关系数（ICC）进行分析。每种病毒在CT和VT中的HIV复制情况相似，但HIV-1在CT和VT中的p24水平分别比HIV-1高1.5个对数和0.9个对数（p < .001），这与HIV-1相对RNA拷贝数相关。两种组织中累积的p24和RNA拷贝数在两种病毒中均显示出较低的个体内相关性（HIV-1的ICC≤0.513；HIV-1的ICC≤0.419）。参加过收集生殖器活检样本的既往临床研究适度降低了CT的HIV-1累积p24，但对VT没有影响。为了改进体外挑战试验，应在研究实施前评估病毒在粘膜组织中的复制情况，如果临床试验中包括安慰剂/无治疗组，则不需要基线粘膜组织，并且应避免以前的活检部位。
