Perforin 2 controls unconventional cytokine release from mucosal APC

穿孔素 2 控制粘膜 APC 的非常规细胞因子释放

• 批准号: 10629434
• 负责人: De'Broski R Herbert
• 金额: $ 45.62万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-20 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629434
• 关键词: Acute; Address; Adopted; Airway Disease; Anti-Inflammatory Agents; Antigen-Presenting Cells; Antigens; Automobile Driving; B-Lymphocyte Subsets; Bacteria; Biochemical; Biological Assay; Bone Marrow; Cell Communication; Cell Death; Cell Shape; Cell membrane; Cell physiology; Cells; Cellular biology; Chimera organism; Chronic; Cytolysis; Cytoplasm; Data; Dendritic Cells; Development; Disease; Exposure to; FOXP3 gene; Family; GATA3 gene; Genes; Genetic Transcription; Growth Factor; Helminths; Human; IFNAR1 gene; IRF4 gene; ITGAX gene; Immunity; Immunosuppression; Impairment; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza A virus; Inhalation; Innate Immune Response; Interferons; Interleukin-1; Interleukins; Location; Macrophage; Mass Spectrum Analysis; Mediating; Mediator; Membrane; Molecular; Mucous Membrane; Mus; Myelogenous; N-terminal; Nasal Polyps; Necrosis; Operative Surgical Procedures; Parasites; Pathway interactions; Patients; Peptide Signal Sequences; Phagolysosome; Population; Protein Secretion; Proteins; Reagent; Regulatory T-Lymphocyte; Reporting; Respiratory Mucosa; Role; Shapes; Signal Transduction; Specimen; T cell response; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic Intervention; Tissues; VDAC1 gene; Virus; Virus Diseases; Work; adaptive immune response; airborne allergen; chromatin immunoprecipitation; chronic inflammatory disease; chronic rhinosinusitis; cytokine; draining lymph node; experimental study; helminth infection; immunoregulation; mass spectrometric imaging; microbicide; mouse model; mutant; novel; pathogen; perforin 2; prevent; programs; protein aminoacid sequence; public health relevance; receptor; respiratory infection virus; respiratory pathogen; respiratory virus; response; single-cell RNA sequencing; theories; trafficking; transcription factor

Perforin 2 controls unconventional cytokine release from mucosal APC Project Summary How professional antigen presenting cell (APC) populations focally deliver cytokines to T cells for shaping the pro-inflammatory vs. anti-inflammatory status of mucosal tissue remains incompletely understood. In particular, cytokines that lack N-terminal peptide sequence such as the IL-1 family cytokine IL-33 can't access conventional protein secretion pathways, which has led to the prevailing view that cell death is responsible for IL-33 release. This project is built upon an exciting set of preliminary data demonstrating that mucosal conventional dendritic cell (cDC) subsets, in both humans and mice, express the transmembrane pore-forming protein Perforin-2, which at least in mice, facilitates IL-33 secretion. While in human cDC, we find Perforin-2 expression primarily in an interferon regulatory factor 4 (IRF4) subset indicating the cDC2 lineage, in mouse cDC, we find Perforin-2 in the CD103+ cDC1 subset known to express the transcription factors Irf8 and Batf3. Irrespective of this lineage distinction, both human and mouse CD11c+ cells in the respiratory mucosa contain cytoplasmic IL-33 protein. Our data demonstrate that inhibition of Perforin-2 activity prevents IL-33 release from cDC and inhibits the proliferative expansion of a poorly understood ST2+GATA3+Foxp3+Treg subset. Given that Perforin-2 has been shown to also regulate Type 1 IFN signaling through controlling IFNAR responsiveness, we propose an important regulatory mechanism exists in humans and mice that is dependent upon mucosal APC that express Perforin-2. This project tests the central hypothesis that APC require Perforin-2 as an inducible plasma membrane conduit for unconventional cytokine delivery at the mucosal interface. Three specific aims (SA) will be investigated. SA1 will determine whether Perforin-2+ APC predict Treg subset abundance in sinonasal mucosa and define the transcriptional landscape of Perforin-2+ APC. SA2 will define the Perforin-2 domains required for IL-33 release, the diversity of Perforin-2-dependent secreted molecules, and the requisite intracellular trafficking machinery responsible for Perforin-2 plasma membrane localization during APC-T cell interactions. SA3 will directly test whether cDC1 and/or cDC2 subsets preferentially use Perforin-2 for driving pathogen-specific T cell responses in mouse models of respiratory virus or helminth infection. Taken together, this MIST project stands to uncover a new paradigm for understanding how cDC instruct immunity within the respiratory mucosa.

穿孔素 2 控制粘膜 APC 的非常规细胞因子释放 项目概要 专业抗原呈递细胞 (APC) 群体如何将细胞因子集中递送至 T 细胞以塑造 粘膜组织的促炎与抗炎状态仍不完全清楚。尤其， 缺乏N端肽序列的细胞因子（例如IL-1家族细胞因子IL-33）无法访问常规细胞因子 蛋白质分泌途径，这导致人们普遍认为细胞死亡是 IL-33 释放的原因。 该项目建立在一组令人兴奋的初步数据之上，这些数据表明粘膜传统树突状 人类和小鼠的细胞 (cDC) 亚群都表达跨膜孔形成蛋白 Perforin-2，该蛋白 至少在小鼠中，促进 IL-33 分泌。在人类 cDC 中，我们发现 Perforin-2 表达主要在 干扰素调节因子 4 (IRF4) 子集表明 CDC2 谱系，在小鼠 CDC 中，我们在 CD103+ cDC1 子集已知表达转录因子 Irf8 和 Batf3。与这个血统无关 区别在于，人和小鼠呼吸道粘膜中的 CD11c+ 细胞都含有细胞质 IL-33 蛋白。 我们的数据表明，抑制 Perforin-2 活性可阻止 cDC 释放 IL-33，并抑制 一个鲜为人知的 ST2+GATA3+Foxp3+Treg 子集的增殖扩张。鉴于 Perforin-2 已 研究表明还可以通过控制 IFNAR 反应来调节 1 型 IFN 信号传导，因此我们提出了一个重要的 人类和小鼠中存在的调节机制依赖于表达 Perforin-2 的粘膜 APC。 该项目测试了 APC 需要 Perforin-2 作为诱导质膜的中心假设 在粘膜界面输送非常规细胞因子的导管。三个具体目标（SA）将是 调查了。 SA1 将决定 Perforin-2+ APC 是否预测鼻窦粘膜中 Treg 子集的丰度 并定义 Perforin-2+ APC 的转录景观。 SA2 将定义所需的 Perforin-2 域 IL-33 释放、穿孔素 2 依赖性分泌分子的多样性以及必要的细胞内运输 在 APC-T 细胞相互作用期间负责 Perforin-2 质膜定位的机制。 SA3将 直接测试 CDC1 和/或 CDC2 子集是否优先使用 Perforin-2 来驱动病原体特异性 T 细胞 呼吸道病毒或蠕虫感染小鼠模型的反应。总而言之，这个 MIST 项目代表 揭示一个新的范例来理解 CDC 如何指导呼吸道粘膜内的免疫。