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

项目。

• 批准号: 7685067
• 负责人: THOMAS S MCCORMICK
• 金额: $ 60.19万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7685067
• 关键词: Address; Adhesions; African American; Anatomic Sites; Anatomy; Anti-Retroviral Agents; Apoptosis; Benchmarking; Binding; Biological Process; CXCR4 gene; Cell Communication; Cell Death; Cell Proliferation; Cells; Chemotaxis; Chronic; Clinical; Coculture Techniques; Communities; Complication; Cytoplasmic Tail; Data; Defensins; Dendritic Cells; Environment; Epidemiology; Epithelial; Epithelial Cells; Epithelium; Event; Exhibits; Exposure to; Female; Future; Genital system; Growth; HIV; HIV Infections; HIV Seropositivity; HIV-1; Heterosexuals; Highly Active Antiretroviral Therapy; Human; Human Papillomavirus; Immature Monocyte; Immune; Immune response; Immunocompromised Host; Immunologic Deficiency Syndromes; In Vitro; Individual; Infection; Inflammatory; Intervention; Leukocyte L1 Antigen Complex; Lichen Planus; Ligands; Location; Messenger RNA; Molecular Profiling; Mucosal Immunity; Mucous Membrane; Mutation; Normal tissue morphology; Nucleosides; Oral; Oral Manifestations; Oral candidiasis; Oral cavity; Oral mucous membrane structure; Oropharyngeal; Patients; Phenotype; Phosphorylation; Play; Population; Predisposition; Production; Property; Protease Inhibitor; Protein Analysis; Proteins; Proteome; Proteomics; Psoriasis; Recruitment Activity; Reverse Transcriptase Inhibitors; Risk; Role; Route; SLPI gene; Sampling; Signal Transduction; Site; Skin; Spottings; Stromal Cell-Derived Factor 1; Syndrome; Testing; Therapeutic; Tissues; Translating; Undifferentiated; Vagina; Viral; Vulnerable Populations; Wound Healing; acronyms; antimicrobial peptide; arrestin3; base; beta-Defensins; beta-defensin 3; beta-defensin-2; chemokine; cohort; cytokine; desensitization; insight; interest; keratinocyte; male; monocyte; oral cavity epithelium; oral wart; protein expression; protein profiling; receptor internalization; rectal; research study; response; steroid hormone; transmission process

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.

我们以前已经证明，口腔上皮细胞产生人β防御素-2和-3 (hBD2，3），对于大多数人来说，在炎症条件下通常可诱导的先天免疫分子 在口腔上皮中，内源性水平较高。应用2D-DIGE评估人口腔粘膜的 来自弱势群体（主要是非洲人）的HIV+和HIV-个体的上皮细胞 美国人），我们确定了153个感兴趣的蛋白质; 137个（-90%）下调，16个下调。 与对照相比，在从HIV阳性个体获得的样品中上调。有趣的是，在 就其生物学功能和意义而言，蛋白质谱与细胞死亡一致 细胞凋亡蛋白是最多的，其次是细胞增殖蛋白和免疫调节蛋白。 反应蛋白，这表明细胞和先天免疫机制可能会改变， 艾滋病毒感染的结果。了解上皮细胞在HIV感染中的作用， 来自不同解剖结构的上皮组织的固有差异敏感性特性 位置感兴趣。我们建议（I）比较人口腔上皮细胞的蛋白质组学谱 （HOEC）、女性生殖道上皮细胞（FGTEC）和皮肤来源的上皮细胞（SDEC） 从HIV+和HIV-受试者，并检查这些上皮细胞中的蛋白质谱， 用HIV或HAART疗法进行攻击;（II）比较先天免疫应答和细胞免疫应答 基线和HIV激发后HOEC、FGTEC和SDEC中的分子， HAART治疗以及在从疣（口腔和阴道）组织获得的上皮细胞中;（III） 确定HOEC、FGTEC或SDEC的蛋白质组反应是否因共孵育而改变 用免疫衍生的未分化和/或分化的细胞在暴露于人β 防御素。我们假设β-防御素和其他抗菌肽在大肠杆菌中升高， 组织经历生长和高增殖率，先天免疫水平升高， 分子可以导致较少的病毒通过粘膜屏障传播。此外，口服 HIV感染的并发症可以通过HIV治疗如HAART或 抗微生物肽的内源性水平。通过研究生殖器上皮细胞中的蛋白质谱， 非常容易受HIV转胞作用和感染部位，并将其与口腔和皮肤进行比较 粘膜，将获得新的见解，然后可以在未来转化为促进 保护脆弱的粘膜屏障。