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

项目。

基本信息

批准号：
8254424
负责人：
THOMAS S MCCORMICK
金额：
$ 31.32万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
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/8254424
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

项目摘要

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.

我们之前已经证明口腔上皮细胞产生人类β防御素-2和-3 (hBD2, 3)，对于大多数人来说，先天免疫分子通常在炎症条件下可诱导上皮细胞，口腔上皮细胞内源性水平较高。使用 2D-DIGE 评估人类口腔来自弱势群体（主要是非洲人）的 HIV+ 和 HIV- 个体的上皮细胞美国人），我们鉴定了 153 种感兴趣的蛋白质； 137 个 (-90%) 被下调，16 个被下调与对照相比，从 HIV 阳性个体获得的样本中表达上调。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）数量最多，其次是细胞增殖蛋白以及免疫学蛋白反应蛋白，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 感染中的作用源自不同解剖结构的上皮组织固有的不同敏感性特性地点是人们感兴趣的。我们建议（I）比较人类口腔上皮细胞的蛋白质组谱（HOEC）、女性生殖道上皮细胞（FGTEC）和皮肤源性上皮细胞（SDEC）来自 HIV+ 和 HIV- 受试者，并在体外检查这些上皮细胞中的蛋白质谱 HIV 或 HAART 治疗的挑战；（二）先天免疫反应与细胞免疫反应的比较 HOEC、FGTEC 和 SDEC 在基线和 HIV 攻击后的分子 HAART 治疗以及从疣（口腔和阴道）组织获得的上皮细胞治疗；（三）确定 HOEC、FGTEC 或 SDEC 的蛋白质组反应是否因共孵育而改变接触人类β后免疫衍生的未分化和/或分化细胞防御素。我们假设β防御素和其他抗菌肽在组织正在生长和高增殖率，并且先天免疫水平升高分子可以减少病毒通过粘膜屏障的传播。另外，口服 HIV 感染的并发症可能会通过 HAART 等 HIV 治疗方法或治疗方法的差异而改变抗菌肽的内源水平。通过研究生殖器上皮细胞中的蛋白质谱，该部位极易受到 HIV 胞吞作用和感染，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。