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

项目。

基本信息

批准号：
8070379
负责人：
THOMAS S MCCORMICK
金额：
$ 29.24万
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/8070379
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 Mucous Membrane Mutation Normal tissue morphology Nucleosides Oral Oral Manifestations Oral candidiasis Oral cavity Oral mucous membrane structure Oropharyngeal Patients Phenotype Phosphorylation Play 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.

我们以前已经证明，口腔上皮产生人β辩蛋白-2和-3 （HBD2，3），大多数炎症条件下通常诱导的先天免疫分子上皮，口腔上皮的较高内源水平。使用人类口服的2D-DIGE评估来自艾滋病毒+和艾滋病毒的上皮细胞来自脆弱人群（主要是非洲人美国人），我们确定了153种感兴趣的蛋白质； 137（-90％）下调，16个在从HIV阳性个体与对照的样本中上调。有趣的是，在其生物学功能和意义的术语，与细胞死亡一致的蛋白质谱（凋亡）是最多的，其次是细胞增殖蛋白以及免疫学反应蛋白，表明细胞和先天免疫机制都可以改变为 HIV感染的结果。了解上皮在艾滋病毒感染中扮演的作用源自各种解剖学的上皮组织的固有差异敏感性位置令人感兴趣。我们建议（i）比较人口腔上皮细胞的蛋白质组学特征（HOEC），女性生殖器轨道上皮细胞（FGTEC）和皮肤衍生的上皮细胞（SDEC）来自HIV+和HIV受试者，并在体外检查这些上皮的蛋白质谱艾滋病毒或HAART治疗的挑战；（ii）比较先天和细胞免疫反应基线时HOEC，FGTEC和SDEC之间的分子，以及艾滋病毒和艾滋病毒的挑战 HAART治疗以及从疣（口腔和阴道）组织获得的上皮细胞中；（iii）确定HOEC，FGTEC或SDEC是否通过共孵化改变了蛋白质组学反应暴露于人类beta后，具有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经历生长和高增殖率的组织以及先天免疫的水平升高分子可能导致通过粘膜屏障的病毒传播较少。另外，口头艾滋病毒感染的并发症可能会因HAART等艾滋病毒治疗剂而改变内源性抗菌肽的水平。通过研究生殖器上皮中的蛋白质谱，A 部位极易受到HIV trancytosis和感染的影响，并将其与口腔和皮肤进行比较粘膜，将获得新的见解，然后将来可以将其翻译成促进在脆弱的粘膜壁垒中进行保护。