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.

项目总结/摘要 我们研究的新证据表明，由独特的16 S rDNA，可能与健康皮肤（核糖体型PH）或痤疮（核糖体型PA）有关。我们建议 研究核糖型PH和PA诱导Th 1与Th 17介导的免疫机制的性质 应答本研究中使用的痤疮丙酸杆菌核糖体型PH和PA是最普遍的，包括三种 每种PH和PA菌株都在健康皮肤的个体中富集或与痤疮相关 患者组。我们将测试我们的中心假设，即痤疮丙酸杆菌核糖型触发T细胞介导 不同的免疫反应，这要么有助于痤疮的发病机制，或导致保护性免疫反应。 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 DermDB以深入了解T细胞分化的机制。具体来说，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签名，以确定决定免疫的网络。 T细胞分化为功能性Th 1和Th 17细胞亚群。我们还将确定机制， 包括PH和PA诱导T细胞分化的细胞因子。虽然痤疮丙酸杆菌的基因组已经被 测序，毒力因子和激活免疫细胞的特异性配体尚不清楚。我们有 先前证明Toll样受体-2（TLR 2）介导抗原与细胞因子之间的相互作用， 呈递细胞和痤疮丙酸杆菌。负责这些相互作用的细菌配体是潜在的 治疗靶点，但目前尚不清楚。在目标2中，我们建议使用生物化学和蛋白质组学 鉴定来自PH和PA的细菌配体的技术，其导致在 目的1，利用siRNA敲低特异性病原体识别受体（PRR），鉴定特异性病原体识别受体， 识别PH和PA以诱导指示适应性免疫的先天性细胞因子。在Aim中生成的数据 2将有助于鉴定与先天PRR相互作用的PH和PA配体，并提供新的见解 Th 1和Th 17细胞的免疫生物学。在目标3中，我们将鉴定Th 1与Th 2的表型和功能。 由痤疮丙酸杆菌核糖型PH和PA诱导的Th 17细胞。使用目标1中概述的信息学方法， 在潜在的新Th 17基因网络中鉴定了IL-26。Th 17的一个未知功能- 衍生的IL-26作为直接的抗菌剂和DNA感应先天免疫的激活剂， 报道在这里，我们将确定编码蛋白对痤疮丙酸杆菌的抗菌机制，包括 IL-26在体外CFU测定中的表达，并通过测定痤疮皮肤样本中IL-26表达细胞的频率， 免疫组织学我们的调查机制，通过不同的核糖体型的一个细菌 触发不同的免疫反应将具有广泛的适用性，以了解微生物组-宿主免疫 反应相互作用以及识别免疫干预的新靶点。总的来说，K 01训练 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。