Kordy Kattayoun, Elliott Julie, Tanner Karen, Johnson Elizabeth J, McGowan Ian M, Anton Peter A
1 Children's Hospital of Los Angeles , Los Angeles, California.
2 Department of Medicine, Center for HIV Prevention Research, UCLA AIDS Institute , David Geffen School of Medicine at UCLA, Los Angeles, California.
AIDS Res Hum Retroviruses. 2018 May;34(5):459-466. doi: 10.1089/AID.2017.0118. Epub 2018 Feb 21.
To determine whether human whole semen (WS) and seminal plasma (SP) either previously frozen or freshly acquired altered ex vivo infectibility of human colonic explants or was associated with histology or toxicity changes, which may influence mucosal HIV-1 transmission in vivo. Pooled human semen samples were freshly obtained from study volunteers (never frozen) and from commercial sources (frozen/thawed). Endoscopically acquired rectal biopsies were evaluated for toxicity following titered ex vivo WS/SP exposure by histological grading and by MTT assay. The ex vivo HIV-1 biopsy challenge model was used to evaluate effects of exposure to either previously frozen or freshly acquired WS/SP on HIV infectibility at a range of viral inocula (10-10 TCID). To evaluate the effects at lower viral inocula of HIV-1 (10-10), experiments in the presence or absence of WS/SP were also performed utilizing TZM-bl cells. MTT assays and histological scoring demonstrated no tissue degradation of biopsies when exposed for 2 h to concentrations of 10% or 100% of either fresh or previously frozen WS/SP. Ex vivo biopsy HIV-1 challenge experiments showed no differences in the presence of freshly acquired or previously frozen/thawed WS/SP compared with control; no differences were seen with lower infectious titers on TZM-bl cells. Within the limits of assay sensitivity and variability, these data show no toxicity or significant enhancement of HIV-1 infectibility of human rectal mucosa using the colorectal explant model with either pooled fresh or frozen/thawed nonautologous human semen.
为了确定先前冷冻或新鲜获取的人类全精液（WS）和精浆（SP）是否会改变人类结肠外植体的体外感染性，或者是否与组织学或毒性变化相关，而这些变化可能会影响体内黏膜HIV-1传播。从研究志愿者（从未冷冻）和商业来源（冷冻/解冻）新鲜获取混合的人类精液样本。通过组织学分级和MTT测定法，在体外对分级的WS/SP进行滴定暴露后，对内窥镜获取的直肠活检组织进行毒性评估。使用体外HIV-1活检挑战模型，评估暴露于先前冷冻或新鲜获取的WS/SP对一系列病毒接种量（10-10 TCID）下HIV感染性的影响。为了评估在较低HIV-1病毒接种量（10-10）下的影响，还利用TZM-bl细胞在有或无WS/SP的情况下进行了实验。MTT测定和组织学评分表明，当活检组织暴露于新鲜或先前冷冻的WS/SP浓度为10%或100%的环境中2小时时，没有组织降解现象。体外活检HIV-1挑战实验表明，与对照相比，新鲜获取或先前冷冻/解冻的WS/SP的存在没有差异；在TZM-bl细胞上较低感染滴度时也没有差异。在测定灵敏度和变异性的范围内，这些数据表明，使用新鲜或冷冻/解冻的非自体混合人类精液的结肠外植体模型，对人类直肠黏膜没有毒性，也不会显著增强HIV-1的感染性。
