Pyles Richard B, Miller Aaron L, Maxwell Carrie, Dawson Lauren, Richardson-Harman Nicola, Swartz Glenn, O'Neill Cynthia, Walker Cattlena, Milligan Gregg N, Madsen Timothy, Motamedi Massoud, Vargas Gracie, Vincent Kathleen L
Department of Pediatrics, The University of Texas Medical Branch, Galveston, TX, United States.
Office of Clinical Research, The University of Texas Medical Branch, Galveston, TX, United States.
Front Reprod Health. 2021 Dec 7;3:714829. doi: 10.3389/frph.2021.714829. eCollection 2021.
The development of therapies targeted to improve the health of women has utilized direct vaginal delivery as a more effective and less toxic method of protection from HIV and other pathogens. Vaginal applicants and delivery devices that provide sustained effects have been met with increasing acceptability, but the efficacy and toxicity outcomes have not been successfully predicted by preclinical studies and animal modeling. We have explored the utilization of sheep as a model for testing the safety of vaginal applicants and devices based on spatial and structural similarities to the human vagina. As recently noted by the FDA, an additional safety measure is an impact on the vaginal microbiome (VMB) that is known to contribute to vaginal health and influence pathogen susceptibility and drug metabolism. To advance the utility of the sheep vaginal model, we completed a thorough molecular characterization of the ovine VMB utilizing both next-generation sequencing (NGS) and PCR methods. The process also created a custom PCR array to quantify ovine VMB community profiles in an affordable, higher throughput fashion. The results from vaginal swabs (>475 samples) collected from non-pregnant crossbred Dorset and Merino ewes treated with selected vaginal applicants or collected as sham samples established 16 VMB community types (VMB CTs). To associate VMB CTs with eubiosis or dysbiosis, we also completed custom ELISAs for six cytokines identifying IL1B, IL8, TNFa, and CXCL10 as useful markers to support the characterization of ovine vaginal inflammation. The results indicated that , and were common markers of eubiosis (low inflammatory marker expression), and that , and were associated with dysbiosis (high cytokine levels). Utilizing the optimized workflow, we also confirmed the utility of three commonly used vaginal applicants for impact on the VMB and inflammatory state, producing a dataset that supports the recommendation for the use of sheep for testing of vaginal applicants and devices as part of preclinical pipelines.
旨在改善女性健康的疗法开发采用直接阴道给药，作为预防艾滋病毒和其他病原体的一种更有效且毒性更小的方法。提供持续效果的阴道制剂和给药装置已越来越被接受，但临床前研究和动物模型尚未成功预测其疗效和毒性结果。基于与人类阴道在空间和结构上的相似性，我们探索了利用绵羊作为模型来测试阴道制剂和装置的安全性。正如美国食品药品监督管理局最近指出的，另一项安全措施是对阴道微生物群（VMB）的影响，已知其有助于阴道健康并影响病原体易感性和药物代谢。为提高绵羊阴道模型的实用性，我们利用下一代测序（NGS）和聚合酶链反应（PCR）方法对绵羊VMB进行了全面的分子表征。该过程还创建了一个定制的PCR阵列，以一种经济高效、高通量的方式量化绵羊VMB群落谱。从接受选定阴道制剂治疗的非妊娠杂交多塞特和美利奴母羊采集的阴道拭子（>475个样本）或作为假样本采集的结果确定了16种VMB群落类型（VMB CTs）。为了将VMB CTs与正常微生物群或失调微生物群相关联，我们还完成了针对六种细胞因子的定制酶联免疫吸附测定（ELISA），确定白细胞介素1β（IL1B）、白细胞介素8（IL8）、肿瘤坏死因子α（TNFa）和CXC趋化因子配体10（CXCL10）为支持绵羊阴道炎症特征化的有用标志物。结果表明，[此处原文缺失具体内容]、[此处原文缺失具体内容]和[此处原文缺失具体内容]是正常微生物群（低炎症标志物表达）的常见标志物，而[此处原文缺失具体内容]、[此处原文缺失具体内容]和[此处原文缺失具体内容]与失调微生物群（高细胞因子水平）相关。利用优化的工作流程，我们还证实了三种常用阴道制剂对VMB和炎症状态的影响，生成了一个数据集，支持将绵羊用于测试阴道制剂和装置作为临床前流程一部分的建议。
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