Toll-Like Receptors and Microbicides

先天性免疫反应是防止性传播疾病(STD)的第一道防线。 先天反应的启动是基于对病原体相关分子模式的识别 宿主蛋白称为Toll样受体(TLRs)。TLR激动剂已被确定并已被利用 用于在疫苗佐剂的背景下调节人类的免疫反应。在这个项目中，我们 建议检验TLR激动剂将被证明是一类新型局部杀微生物剂的假设 防止性传播疾病。局部杀微生物剂是旨在防止性传播疾病的产品。 经阴道或直肠使用的杀微生物剂为女性提供了一个优势，即它们可能是“女性控制的”。 术语杀微生物剂意味着产品杀死微生物；这在某种程度上是误导的，因为 杀菌剂的作用方式多种多样，包括破坏病原体的膜或包膜， 阻断对传染性至关重要的受体-配体相互作用，抑制细胞内或细胞外 病原体的复制，或通过改变阴道环境或增强局部免疫反应 降低敏感度。作为概念证明，我们已经确定了TLR9激动剂作为杀微生物剂的用途 这对小鼠和豚鼠模型中的HSV-2生殖器感染有效。这位杀微生物剂候选人 在给药前2天至后6小时内单次阴道给药有效 用HSV-2对阴道进行挑战。为了扩展我们的发现并检验我们的假设，在目标1中，我们将确定 TLR基因在人阴道和宫颈上皮细胞中的典型表达模式 TLR激动剂，靶向共同表达的TLRs，然后将评估其在初级 这些细胞的培养抗HSV-2和沙眼衣原体感染。TLR激动剂将优先用于 根据体外结果进行评估，然后在小鼠模型中评估杀微生物剂的潜力 单纯疱疹病毒2型和沙眼衣原体阴道感染的研究2.那些显示出最强杀菌剂的化合物 潜力将在目标3中进行机械评估。完成研究将提供大量 关于阴道粘膜的先天免疫反应的信息，并将识别TLR激动剂 杀微生物剂候选人。

innate immune responses are the first line of defense against sexually transmitted diseases (STDs). Initiation of the innate response is based upon recognition of pathogen-associated molecular patterns by host proteins known as toll-like receptors (TLRs). TLR agonists have been identified and have been utilized for modulation of immune responses in humans in the context of vaccine adjuvants. In this project, we propose to test the hypothesis that TLR agonists will prove to be a novel class of topical microbicides providing protection against STDs. Topical microbicides are products designed to protect against STDs. Used vaginally or rectally, microbicides offer women the advantage that they may be "female-controlled". The term microbicide implies that the product kills microorganisms; this is somewhat misleading since microbicides may work in a variety of ways including disrupting the pathogen's membrane or envelope, blocking the receptor-ligand interactions essential for infectivity, inhibiting the intra- or extra-cellular replication of the pathogen, or by altering the vaginal environment or enhancing local immune responses to reduce susceptibility. As proof of concept, we have established the utility of a TLR9 agonist as a microbicide that is effective against HSV-2 genital infections in mice and guinea pig models. This microbicide candidate was effective when applied as a single intravaginal dose within the time frame of 2d prior through 6h after vaginal challenge with HSV-2. To extend our findings and test our hypothesis, in Aim 1 we will determine the typical pattern of TLR gene expression in human vaginal and cervical epithelial cells and select a set of TLR agonists that target the commonly expressed TLRs that then will be assessed for activity in primary cultures of these cells against HSV-2 and C. trachomatis infection. TLR agonists will be prioritized for evaluation based on the in vitro results and then will be evaluated for microbicide potential in mouse models of HSV-2 and C. trachomatis vaginal infections in Aim 2. Those compounds that show greatest microbicide potential will be evaluated mechanistically in Aim 3. Completion of the studies will provide substantial information about the innate immune responses of the vaginal mucosa and will identify TLR agonist microbicide candidates.