The oral mucosal proteome: perturbation in HIV infection and Candida co infection

描述(由申请者提供)：这项多学科的申请支持以患者为中心的研究与基于科学的疾病病因调查相结合的战略。总体目标是了解明显保护口腔上皮免受艾滋病毒感染的机制。长期目标是了解免疫受损宿主共病的基础，从而提供更有效的治疗方法。我们假设HIV直接影响口腔上皮蛋白质组，导致促进共病原体念珠菌定植和感染的变化。其具体目标是：1)充分描述保护口腔粘膜免受艾滋病毒和念珠菌感染的口腔上皮成分。利用口腔和阴道上皮模型，以及人类和猕猴样本，将用流式细胞术研究HIV和念珠菌的典型受体。然后将对存在或不存在HIV和念珠菌的上皮蛋白质组进行全面的表征，以确定这些粘膜中对HIV和念珠菌感染的保护性或易感性因素。唾液将被应用到模型中，以调查口服液中存在的可能抑制HIV或念珠菌感染的先天分泌因子的作用。将进行转录谱分析、定量RT-PCR和Luminex蛋白分析来支持蛋白质组数据。2)研究人类上皮细胞在HIV和念珠菌感染过程中所诱发的天然TLR相关蛋白质组反应。在口腔和阴道上皮模型中，首先将利用蛋白质组学研究HIV和C.a/b/Cans诱导的上皮细胞因子、趋化因子、TLR1-10和信号通路的表达谱。然后，将使用转录谱、实时RT-PCR、共聚焦和免疫电子显微镜以及siRNAi来充分表征PMN依赖的TLR介导的预防口腔白念珠菌感染的保护机制。相关性：艾滋病毒/艾滋病最常见的传播途径是粘膜。在某些情况下，口腔组织暴露在艾滋病毒中，但很少被感染。了解口腔明显免受感染的机制可能会导致开发出保护其他粘膜组织(如阴道)免受病毒感染和传播的治疗方法。这也可能导致治疗与艾滋病毒相关的混合感染，如口腔画眉。

DESCRIPTION (provided by applicant): This multidisciplinary application supports a strategy of combining patient-oriented research with the science based investigation of disease causation. The overall aim is to understand the mechanisms that apparently protect the oral epithelium from HIV infection. The long-term objective is to understand the basis of copathogenicity in the immunocompromised host leading to the delivery of more effective therapies. We hypothesize that HIV directly affects the oral epithelial proteome, leading to alterations that promote colonization and infection of the co-pathogen Candida. The specific aims are: 1) To fully characterize the constituents of the oral epithelium that protects the oral mucosa from HIV and Candida infection. Using oral and vaginal epithelial models, and human and macaque samples, the canonical receptors for both HIV and Candida will be investigated by flow cytometry. The epithelial proteome in the presence or absence of HIV and Candida will be then be fully characterized in order to identify protective or susceptibility factors to HIV and Candida infection in these mucosae. Saliva will be applied to the models to investigate the contribution of innate secretory factors present in oral fluids that may inhibit HIV or Candida infection Transcript profiling, quantitative RT-PCR, and Luminex protein assays will be performed to support the proteomic data. 2) To characterize the innate TLR-associated proteomic response elicited by human epithelium during HIV and Candida infection. The HIV- and C. a/b/cans-induced epithelial cytokine, chemokine, TLR1-10 and signaling pathway expression profiles will first be characterized in oral and vaginal epithelial models using proteomics. The PMN-dependent TLR-mediated protection mechanism against oral C. albicans infection will then be fully characterized using transcript profiling, real-time RT-PCR, confocal and immunoelectron microscopy, and siRNAi. Relevance: HIV/AIDS is most commonly transmitted through mucosal membranes. In some circumstances, the oral tissues are exposed to HIV but rarely become infected. Understanding the mechanism by which the oral cavity is apparently protected from infection could lead to the development of therapies to protect other mucosal tissues such as the vagina from infection and transmission of the virus. This could also lead to therapies for the treatment of the co-infections associated with HIV such as oral Thrush.