Proj. 3 - Proteomic Response of Epithelial Cell Interactions with HIV

我们先前已经证明口腔上皮细胞产生人类β防御素-2和-3。 (hBD2，3)，天然免疫分子，通常在炎症条件下诱导 口腔上皮细胞，内源性水平较高。应用2D-DGE评价人的口腔功能 来自HIV+的上皮细胞和来自脆弱人群(主要是非洲人)的HIV个体 ，我们确定了153个感兴趣的蛋白质；137个(-90%)下调，16个下调 与对照组相比，从HIV阳性个体获得的样本中上调。有趣的是，在 它们的生物学功能和意义，与细胞死亡相一致的蛋白质谱 (细胞凋亡)数量最多，其次是细胞增殖蛋白和免疫学 反应蛋白，表明细胞和先天免疫机制可能被改变为 是艾滋病毒感染的结果。了解上皮细胞在HIV感染和感染中的作用 不同解剖来源的上皮组织的固有差异易感性 地点很有意义。我们建议(I)比较人类口腔上皮细胞的蛋白质组学特征 (HOECs)、女性生殖道上皮细胞(FGTECs)和皮肤来源上皮细胞(SDECs) 从HIV+和HIV-受试者身上提取蛋白质，并在体外检测这些上皮细胞的蛋白质谱 挑战HIV或HAART疗法；(Ii)比较先天免疫反应和细胞免疫反应 HOECs、FGTECs和SDECs在基线和感染HIV和 HAART治疗以及从疣(口腔和阴道)组织中获得的上皮细胞； 确定HOECs、FGTECs或SDECs的蛋白质组反应是否通过共孵育而改变 在暴露于人β后，免疫来源的未分化和/或已分化细胞 防御素。我们假设β防御素和其他抗菌肽在 正在经历生长和高增殖率以及先天免疫水平升高的组织 分子可能会减少病毒通过粘膜屏障的传播。此外，口述 HIV感染的并发症可以通过HAART等HIV疗法或通过改变 内源性抗菌肽水平。通过研究生殖器上皮细胞中的蛋白质图谱， 极易感染艾滋病毒的部位，并将其与口腔和皮肤进行比较 粘膜，将获得新的见解，这些见解将在未来转化为促进 对脆弱的粘膜屏障的保护。

We have previously demonstrated that oral epithelia produces human beta defensin-2 and-3 (hBD2, 3), innate immune molecules ordinarily inducible under inflammatory conditions for most epithelia, at higher endogenous levels in oral epithelia. Using 2D-DIGE assessment of human oral epithelial cells from HIV+ and HIV- individuals from vulnerable populations (mainly African Americans), we identified 153 proteins of interest; 137 (-90%) were down-regulated and 16 were up-regulated in samples obtained from HIV positive individuals versus control. Interestingly, in terms of their biological functions and significance, protein profiles consistent with cell death (apoptosis) were the most numerous followed by cell proliferation proteins as well as immunological response proteins, suggesting that both cellular and innate immune mechanisms may be altered as a result of HIV infection. Understanding the role that the epithelium plays in HIV infection and inherent differential susceptibility properties of epithelial tissues derived from various anatomic locations is of interest. We propose to (I) compare proteomic profiles of human oral epithelial cells (HOECs), female genital track epithelial cells (FGTECs) and skin-derived epithelial cells (SDECs) from HIV+ and HIV- subjects and to examine protein profiles in these epithelia following in vitro challenge with HIV or HAART therapy; (II) Compare innate and cellular immune response molecules among HOECs, FGTECs and SDECs at baseline and following challenge with HIV and HAART treatment as well as among epithelial cells obtained from wart (oral and vaginal) tissue; (III) Determine if proteomic response from HOECs, FGTECs or SDECs are altered by co-incubation with immune-derived undifferentiated and/or differentiated cells following exposure to human beta defensins. We hypothesize that beta defensins, and other antimicrobial peptides are elevated in tissue undergoing growth and high proliferation rates and that elevated levels of innate immune molecules could result in less viral transmission through the mucosal barrier. Additionally, oral complications of HIV infection may be altered by HIV therapeutics such as HAART or differences in endogenous levels of antimicrobial peptides. By studying protein profiles in the genital epithelia, a site extremely susceptible to HIV trancytosis and infection, and comparing them to oral and skin mucosa, new insights will be gained which could then be translated in the future into promoting protection in vulnerable mucosal barriers.