The nature of Propionibacterium acnes-induced inflammatory immune responses in acne vulgaris

痤疮丙酸杆菌诱导寻常痤疮炎症免疫反应的性质

• 批准号: 9293615
• 负责人: George W Agak
• 金额: $ 16.74万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-12 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9293615
• 关键词: Acne; Acne Vulgaris; Affect; Antigen-Presenting Cells; Area; Atopic Dermatitis; Biochemical; Biological Assay; CD4 Positive T Lymphocytes; Cells; Clinical; DNA; Data; Databases; Disease; Epithelial Cells; Frequencies; Genes; Genome; Helper-Inducer T-Lymphocyte; Human; Immune; Immune Targeting; Immune response; Immunobiology; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Response; Informatics; Interleukin-10; Interleukin-17; Investigation; Lead; Ligands; Mediating; Microbe; Modeling; Myelogenous; Natural Immunity; Nature; Outcome; Pathogenesis; Patients; Pattern; Phenotype; Play; Production; Propionibacterium acnes; Proteins; Proteomics; Psoriasis; Psychological Impact; Public Health; Recombinant DNA; Reporting; Research Personnel; Ribotypes; Role; Skin; Small Interfering RNA; Source; Specimen; Symbiosis; T cell differentiation; T-Lymphocyte; T-Lymphocyte Subsets; TLR2 gene; Techniques; Testing; Therapeutic Intervention; Time; Training Programs; Virulence Factors; adaptive immunity; antimicrobial; antimicrobial drug; antimicrobial peptide; cell type; commensal microbes; cytokine; genetic signature; human disease; immunological intervention; insight; interleukin-22; knock-down; microbiome; novel; pathogen; receptor; response; self esteem; skills; skin disorder; targeted treatment; therapeutic target; tool

PROJECT SUMMARY/ABSTRACT New evidence from our studies demonstrates that specific P. acnes ribotypes, distinguished by unique 16S rDNA, may be associated with either healthy skin (ribotype PH) or acne (ribotype PA). We propose to investigate the nature of immune mechanisms by which ribotypes PH and PA induce Th1 vs. Th17-mediated responses. The P. acnes ribotypes PH and PA to be used in this study are the most prevalent and include three each of PH and PA strains that have been enriched in individuals with healthy skin or associated with the acne patient group. We will test our central hypothesis that that P. acnes ribotypes trigger T cells that mediate distinct immune responses, which either contribute to the pathogenesis of acne or lead to a protective mechanism. In Aim 1, we will use informatics, taking advantage of the publicly available skin database DermDB to gain insight into the mechanisms of T cell differentiation. Specifically, using RNAsequencing, we will derive gene signatures of PH and PA-activated innate immune cells, to identify the networks that determine T cell differentiation into functional Th1 and Th17 cell subsets. We will also determine the mechanism(s), including cytokine, by which PH and PA induce T cell differentiation. Although the P. acnes genome has been sequenced, virulence factors and specific ligand(s) that activate immune cells are not clear. We have previously demonstrated that the Toll like receptor-2 (TLR2) mediates the interaction between antigen presenting cells and P. acnes. The bacterial ligand(s) responsible for these interactions are potential therapeutic targets, but are currently unknown. In Aim 2, we propose to use biochemical and proteomic techniques to identify bacterial ligand(s) from PH and PA that lead to differential immune signatures identified in Aim 1, and using siRNA knockdown of specific pathogen recognition receptors (PRR), identify the PRRs that recognize PH and PA to induce the innate cytokines that instruct adaptive immunity. The data generated in Aim 2 will help in the identification of PH and PA ligand(s) that interact with innate PRRs, and provide new insights into the immunobiology of Th1 and Th17 cells. In Aim 3, we will identify the phenotype and function of Th1 vs. Th17 cells induced by P. acnes ribotypes PH and PA. Using the informatics approaches outlined in Aim 1 we have identified IL-26 within the potential new Th17 gene networks. A previously unknown function of Th17- derived IL-26 as a direct antimicrobial agent and activator of DNA-sensing innate immunity was recently reported. Here, we will determine the antimicrobial mechanisms of encoded proteins against P. acnes including IL-26 in in vitro CFU assays, and determine the frequency of IL-26 expressing cells in acne skin specimens by immunohistology. Our investigations of the mechanisms by which distinct ribotypes of a commensal bacterium trigger divergent immune responses will have broad applicability to understanding microbiome-host immune response interactions as well as identify novel targets for immunological intervention. Overall, the K01 training program will allow me to develop the skills and tools needed to become a successful independent investigator.

项目摘要/摘要 我们研究的新证据表明，特定的痤疮假单胞菌核型，由独特的16S区分 RDNA，可能与健康皮肤(核型PH)或痤疮(核型PA)有关。我们建议 研究核糖型PH和PA诱导Th1与Th17介导的免疫机制的本质 回应。本研究中使用的痤疮假单胞菌核型PH和PA是最流行的，包括三种 每一种在健康皮肤或与痤疮有关的人身上富含的PH和PA菌株 患者组。我们将验证我们的核心假设，即痤疮假单胞菌核型触发T细胞，介导 不同的免疫反应，要么导致痤疮的发病，要么导致保护性的 机制。在目标1中，我们将使用信息学，利用公开可用的皮肤数据库 以深入了解T细胞分化的机制。具体地说，使用RNA测序，我们 将获得PH和PA激活的先天免疫细胞的基因特征，以确定决定 T细胞分化为功能性Th1和Th17细胞亚群。我们还将确定机制(S)， 包括细胞因子，PH和PA通过这些细胞因子诱导T细胞分化。尽管痤疮假单胞菌的基因组已经 目前，毒力因子和激活免疫细胞的特异性配体(S)的序列尚不清楚。我们有 先前发现Toll样受体-2(TLR2)介导了抗原与Toll样受体2之间的相互作用 呈现细胞和痤疮假单胞菌。负责这些相互作用的细菌配体(S)是潜在的 治疗靶点，但目前尚不清楚。在目标2中，我们建议使用生化和蛋白质组学 从PH和PA中识别导致差异免疫特征的细菌配体(S)的技术 目的1，利用特异性病原体识别受体(PRR)的siRNA敲除，鉴定可 识别PH和PA以诱导引导适应性免疫的先天细胞因子。在AIM中生成的数据 2将有助于识别与先天性PRR相互作用的PH和PA配体(S)，并提供新的见解 进入Th1和Th17细胞的免疫生物学。在目标3中，我们将确定Th1和Th1的表型和功能。 痤疮假单胞菌核型PH和PA诱导的Th17细胞。使用目标1中概述的信息学方法，我们 已经在潜在的新的Th17基因网络中发现了IL-26。Th17以前未知的功能- 衍生的IL-26作为一种直接的抗菌剂和DNA传感天然免疫的激活剂，最近被认为是 据报道。在这里，我们将确定编码蛋白对痤疮假单胞菌的抗菌机制，包括 IL-26体外CFU检测，并测定痤疮皮肤标本中IL-26表达细胞的频率。 免疫组织学。我们对一种不同核型共生菌的作用机制的研究 触发不同的免疫反应将对理解微生物群-宿主免疫具有广泛的适用性 反应相互作用以及确定免疫干预的新靶点。总体而言，K01培训 该计划将使我能够发展成为一名成功的独立调查员所需的技能和工具。