Department of Pediatrics, University of Texas Medical Branch, Galveston, TX, USA; Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, TX, USA.
Department of Neuroscience & Cell Biology, University of Texas Medical Branch, Galveston, TX, USA; Center for Biomedical Engineering, University of Texas Medical Branch, Galveston, TX, USA.
J Reprod Immunol. 2017 Nov;124:38-43. doi: 10.1016/j.jri.2017.09.014. Epub 2017 Oct 13.
The presence of genital inflammatory responses and a compromised vaginal epithelial barrier have been linked to an increased risk of HIV acquisition. It is important to assure that application of candidate microbicides designed to limit HIV transmission will not cause these adverse events. We previously developed high resolution in vivo imaging methodologies in sheep to assess epithelial integrity following vaginal application of a model microbicide, however characterization of genital inflammation in sheep has not been previously possible. In this study, we significantly advanced the sheep model by developing approaches to detect and quantify inflammatory responses resulting from application of a nonoxynol-9-containing gel known to elicit vaginal irritation. Vaginal application of this model microbicide resulted in foci of disrupted epithelium detectable by confocal endomicroscopy. Leukocytes also infiltrated the treated mucosa and the number and composition of leukocytes obtained by cervicovaginal lavage (CVL) were determined by differential staining and flow cytometry. By 18h post-treatment, a population comprised predominantly of granulocytes and monocytes infiltrated the vagina and persisted through 44h post-treatment. The concentration of proinflammatory cytokines and chemokines in CVL was determined by quantitative ELISA. Concentrations of IL-8 and IL-1β were consistently significantly increased after microbicide application suggesting these cytokines are useful biomarkers for epithelial injury in the sheep model. Together, the results of these immunological assessments mirror those obtained in previous animal models and human trials with the same compound and greatly extend the utility of the sheep vaginal model in assessing the vaginal barrier and immune microenvironment.
生殖器炎症反应和阴道上皮屏障受损与 HIV 感染风险增加有关。重要的是要确保应用旨在限制 HIV 传播的候选杀微生物剂不会引起这些不良反应。我们之前在绵羊中开发了高分辨率体内成像方法，以评估阴道应用模型杀微生物剂后上皮完整性，但是之前无法对绵羊的生殖器炎症进行特征描述。在这项研究中，我们通过开发方法来检测和量化已知会引起阴道刺激的含壬苯醇醚-9 的凝胶阴道应用后的炎症反应，从而显著推进了绵羊模型。这种模型杀微生物剂的阴道应用导致可通过共聚焦内镜检测到的上皮破坏焦点。白细胞也浸润了治疗后的黏膜，通过宫颈阴道灌洗 (CVL) 获得的白细胞数量和组成通过差异染色和流式细胞术确定。在治疗后 18 小时，主要由粒细胞和单核细胞组成的群体浸润阴道，并在治疗后 44 小时持续存在。通过定量 ELISA 测定 CVL 中促炎细胞因子和趋化因子的浓度。在杀微生物剂应用后，IL-8 和 IL-1β 的浓度一直显著增加，表明这些细胞因子是绵羊模型上皮损伤的有用生物标志物。这些免疫学评估的结果与相同化合物的先前动物模型和人体试验结果一致，并极大地扩展了绵羊阴道模型在评估阴道屏障和免疫微环境中的应用。
