Th17 extracellular trap-mediated antimicrobial host defense in acne vulgaris

项目总结 寻常痤疮，或痤疮，是一种毛发皮脂腺单位(PSU)的疾病。粉刺排名第三。 慢性皮肤病是造成残疾和医疗费用的主要原因，也是心理压力的主要原因 在年轻人身上。导致痤疮的一个因素是痤疮杆菌，它是痤疮中的主要细菌物种。 PSU。以痤疮念珠菌为模型研究微生物群与宿主免疫的相互作用 作为回应，我们：1)以前证明IL-17在痤疮皮损中存在，2)最近 研究发现，与痤疮(CA)或健康皮肤(CH)相关的痤疮假丝酵母菌种型对 TH17细胞发育成非抗菌(n-AMTH17)和抗菌(AMTH17)亚群的命运。AMTH17细胞 释放T细胞胞外陷阱(Tet)，直接杀灭痤疮假单胞菌和其他细菌病原体。 迄今为止，痤疮假丝酵母菌系统型别诱发粉刺发作的机制及其生物学影响已有报道。 粉刺是未知的。因此，我们提出的研究是创新的，因为我们将定义免疫 痤疮皮损的景观，提供了对TH17-Tet形成的生物影响的机械性见解 痤疮，并确定新的免疫途径和潜在的生物标记物，可以作为治疗痤疮的目标。 这一点很重要，因为未来可以制定策略来调节TH17的功能。此外，维护 痤疮链霉菌不同系统类型之间的平衡可能代表了一种新的益生菌设计策略。 痤疮链霉菌配体的鉴定将进一步阐明宿主受体参与微生物的特异性。 监测并导致开发新的治疗方法 由生物失调引起的皮肤病。 控制粉刺和其他人类 我们的中心假设是TH17细胞的先天激活导致了抗菌剂的诱导 机制，包括TETS，这有助于宿主防御痤疮假单胞菌和其他细菌。至 阐明这一点，我们将确定AMTH17细胞对痤疮念珠菌的抗菌机制(目标1)， 研究AMTH17细胞在痤疮炎症中的作用(目标2)，并确定诱导痤疮发生的配体 AMTH17分化(目标3)。我们的初步发现支持这样一个前提，即健康的皮肤共生体 对我们免疫系统的教育和我们对病原体的全面防御至关重要。我们的战略将 包括：经典免疫学技术，包括T细胞克隆以剖析免疫效应器功能 这是TH17介导的抗微生物宿主防御的基础；痤疮念珠菌系统型的微生物组测序 确定皮肤微生物群和宿主免疫反应之间的相互作用 影响痤疮的发展；高分辨率时间推移成像，以确定粉刺的释放和机制 Tet形成过程中的染色质动力学；用扫描电子显微镜确定细胞外陷阱 在人类T细胞中形成；以及最先进的单细胞RNA-SEQ实验来计算T细胞基因 并确定痤疮皮损中存在的T细胞类型和免疫回路。更多最新情况- ART方法包括使用高亲和力捕获细胞相互作用，结合质量 基于光谱的蛋白质组学、脂质组学和糖类HPAEC-PAD分析鉴定痤疮念珠菌 配体和内源络合物。这一贡献意义重大，因为我们的研究将对 我们在理解T细胞-抗微生物防御机制方面的概念性进展，并允许主要 可作为治疗靶点的痤疮免疫网络研究进展

PROJECT SUMMARY Acne vulgaris, or acne, is a disease of the pilosebaceous unit (PSU). Acne is ranked third among chronic skin diseases for causing disability and medical expense and is a major cause of psychological stress in young people. One factor contributing to acne is Cutibacterium acnes, the major bacterial species in the PSU. Using C. acnes as a model to study the interaction between the microbiome and the host immune response, we: 1) previously demonstrated the presence of IL-17 in acne skin lesions and 2) recently discovered that C. acnes phylotypes associated with acne (CA) or healthy skin (CH) differentially regulate the fate of TH17 cells to develop into non-antimicrobial (n-AMTH17) and antimicrobial (AMTH17) subsets. AMTH17 cells release T cell extracellular traps (TETs) and directly kill C. acnes and other bacteria pathogens. To date, the mechanisms by which C. acnes phylotypes induce TETs and their biological impact in acne are unknown. Therefore, our proposed research is innovative because we will define the immune landscape of the acne lesions, provide mechanistic insights into the biological impact of TH17-TET formation in acne, and identify novel immune pathways and potential biomarkers that can be targeted for acne therapy. This is important as future strategies could be developed to modulate TH17 function. Additionally, maintaining the balance among the different phylotypes of C. acnes may represent a strategy for novel probiotic design. The identification of C. acnes ligands will further elucidate the specificity of host receptors involved in microbial surveillance and lead to the development of novel therapeutic approaches skin diseases caused by dysbiosis. to control acne and other human Our central hypothesis is that innate activation of TH17 cells leads to induction of antimicrobial mechanisms, including TETs, which contribute to host defense against C. acnes and other bacteria. To elucidate this, we will determine the antimicrobial mechanisms of AMTH17 cells against C. acnes (Aim 1), investigate the role of AMTH17 cells in acne inflammation (Aim 2), and identify the C. acnes ligands that induce AMTH17 differentiation (Aim 3). Our preliminary findings support the premise that healthy skin commensals are critical to the education of our immune system and our overall defense against pathogens. Our strategy will include: classical immunological techniques involving T cell cloning to dissect the immune effector functions that underlie TH17-mediated antimicrobial host defense; microbiome sequencing of C. acnes phylotypes that inhabit donor biopsies to define how interactions within the skin microbiome and the host immune response influences acne development; high-resolution time-lapse imaging to define mechanisms of TET release and chromatin dynamics that occur during TET formation; scanning electron microscopy to delimit extracellular trap formation in human T cells; and state-of-the-art single cell RNA-seq experiments to compute T cell gene signatures and to define the T cell types and immune circuits present in acne lesions. Additional state-of-the- art methodologies include the use of high affinity capture of cellular interactomes, coupled with mass spectrometry-based proteomics, lipidomics and carbohydrate HPAEC-PAD analysis to identify C. acnes ligands and endogenous complexes. This contribution is significant as, our studies will make significant conceptual advances in our understanding of T cell–antimicrobial defense mechanisms, and allow major advances in the understanding of the immune networks in acne that can be targeted for therapy