Systemic coordination of pro-inflammatory immune reactions through dendritic cell-restricted sIL6R biogenesis

通过树突状细胞限制的 sIL6R 生物发生系统协调促炎性免疫反应

• 批准号: 10533314
• 负责人: Daniel Lingwood
• 金额: $ 39.21万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-22 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533314
• 关键词: Adjuvant; Animals; Antibodies; Antibody Response; Antigens; B cell differentiation; B-Lymphocytes; Biogenesis; Biology; Buffers; Cell Line; Cell Lineage; Cell membrane; Cell surface; Cells; Circulation; Complex; Data; Dendritic Cells; Disease; Embryo; Engineering; Evaluation; Failure; Gene Delivery; Genetic; Genetic Polymorphism; Goals; Half-Life; Homeostasis; Human; Humoral Immunities; IL6R gene; Immune; Immune response; Immune signaling; Immunity; Immunization; Immunoglobulins; Immunologic Deficiency Syndromes; Immunologic Receptors; Immunology; Infection; Infiltration; Inflammatory; Influenza; Innate Immune Response; Interleukin 6 Receptor; Interleukin-6; Knock-out; Knockout Mice; Laboratories; Link; Lymphoid; Lymphoid Cell; Mediating; Medical Faculty; Metalloproteases; Mission; Modality; Mus; Natural Immunity; Output; Pathway interactions; Peptide Hydrolases; Phase I Clinical Trials; Plasma Cells; Property; Proteins; Serum; Signal Transduction; Site; Source; Surface; System; T-Lymphocyte; Testing; Therapeutic; Tissues; Transgenic Animals; United States National Institutes of Health; Vaccination; Vaccine Adjuvant; Vaccine Antigen; Vaccines; Viral Vaccines; adaptive immunity; chronic inflammatory disease; combinatorial; conditional knockout; cytokine; defined contribution; druggable target; functional restoration; human disease; immunoreaction; immunosuppressed; improved; in vivo; influenza virus vaccine; medical schools; member; mouse model; novel; prevent; professor; receptor; receptor expression; reverse genetics; targeted treatment; tissue injury; tissue/cell culture; transcription factor; vaccine response

Project Summary / Abstract This is an application by Dr. Daniel Lingwood (PI), and Dr. Alejandro Balazs (Co-I), Assistant Professors at Harvard Medical School and faculty members of the Ragon Institute of MGH, MIT and Harvard. Their laboratories specialize in defining the immunology of antibody responses to vaccines (Lingwood) and in the engineering of immunity through AAV-mediated gene delivery (Balazs). Dr. Lingwood has discovered that antibody responses to vaccination, and pro-inflammatory immune reactions generally, are centrally regulated by novel control axis supplied by conventional dendritic cells (cDC), a single lymphoid lineage defined in mice by expression of the transcription factor Zbtbt46. Dr. Lingwood has found that cDCs are responsible for the in vivo biogenesis of the soluble, circulating form of the IL-6 receptor (sIL6R), which normally captures IL-6 from solution to systemically coordinate its signaling activity. IL-6 is a potent pro-inflammatory cytokine released following immune challenge or tissue injury, and dysregulation of its capture by sIL6R leads to hyperphysiologic IL-6 levels and inflammatory disease. This proposal will test the central hypothesis that biogenesis of sIL6R by cDC forms a centralized control axis that can be modulated to enhance immune defense and restore IL-6 homoeostasis in disease states. In Aim 1, Dr. Lingwood will define the contribution of the metalloproteases ADAM10 and ADAM17 to systemic sIL6R release from the cDC cell surface. These proteases cleave surface IL6R to shed sIL6R from tissue culture cells lines, potentiating a drugable target to modulate IL-6 immune effects. However, ADAM10/17 knockouts are embryonic lethal, preventing mechanistic evaluation of their activity in vivo. Dr. Lingwood has now selectively directed ADAM10 and/or ADAM17 knockouts to the cDC lineage and will use these transgenic animals to define ADAM activity within two cDC- dependent parameters: circulating sIL6R set-point and induction of sIL6R biogenesis after immune challenge. Dr. Lingwood has also discovered that cDC-derived sIL6R, and subsequent trans IL-6 signaling through the sIL6R:IL-6 complex, is necessary for B cell differentiation to antibody secreting plasma cells, and tunes antibody output following immunization with a variety of protein vaccine antigens. In Aim 2, Dr. Lingwood will define how cDC-derived sIL6R links innate and adaptive immunity through this pathway (outside the traditional T cell priming activity of cDC), and more specifically how this can be harnessed as a natural adjuvant principle to improve the efficacy of influenza vaccines. In Aim 3, Dr. Lingwood and Dr. Balazs have developed a AAV- gene delivery platform that rescues sIL6R levels and its IL-6 capture activity in vivo. They will use this system to experimentally define how disease-causing polymorphisms within sIL6R dysregulate IL-6 capture and inflammatory activity in vivo, and then in parallel, the therapeutic sIL6R expression level needed to correct IL-6 homeostasis and restore function therein. Collectively, this proposal will define and exploit newly discovered biology of sIL6R immune-defense to treat human disease, consistent with the goals and mission of the NIH.

项目总结/摘要 这是一份由丹尼尔林伍德博士（PI）和亚历杭德罗巴拉兹博士（Co-I），助理教授在申请 哈佛医学院以及麻省理工学院和哈佛大学Ragon研究所的教员。他们的 实验室专门定义疫苗抗体反应的免疫学（Lingwood）， 通过AAV介导的基因递送工程化免疫（Balazs）。林伍德博士发现 对疫苗接种的抗体反应和一般的促炎免疫反应是由中枢调节的 通过由常规树突状细胞（cDC）提供的新控制轴， 通过转录因子Zbtbt 46的表达。Lingwood博士发现，cDCs是导致 可溶性循环形式的IL-6受体（sIL 6 R）的体内生物发生，其通常从细胞中捕获IL-6， 解决方案，以系统地协调其信号传导活动。IL-6是一种有效的促炎细胞因子， 在免疫攻击或组织损伤后，sIL 6 R对其捕获的失调导致 高生理性IL-6水平和炎性疾病。这一提议将检验中心假设， cDC对sIL 6 R的生物合成形成了一个集中的控制轴，可以通过调节该轴来增强免疫功能。 防御和恢复疾病状态中的IL-6稳态。在目标1中，Lingwood博士将定义 金属蛋白酶ADAM 10和ADAM 17从cDC细胞表面释放全身性sIL 6 R。这些 蛋白酶切割表面IL 6 R以从组织培养细胞系中脱落sIL 6 R，增强可药用靶点， 调节IL-6免疫效应。然而，ADAM 10/17敲除是胚胎致死的，阻止了机制性的 评价其体内活性。Lingwood博士现在选择性地将ADAM 10和/或ADAM 17 cDC谱系的基因敲除，并将使用这些转基因动物来定义两个cDC中的ADAM活性。 依赖性参数：免疫激发后循环sIL 6 R设定点和sIL 6 R生物合成的诱导。 博士Lingwood还发现，cDC衍生的sIL-6 R和随后的反式IL-6信号转导通过 sIL 6 R：IL-6复合物是B细胞分化为分泌抗体的浆细胞所必需的， 用各种蛋白质疫苗抗原免疫后的抗体输出。在目标2中，林伍德博士将 定义cDC衍生的sIL 6 R如何通过该途径（传统免疫途径之外）连接先天性免疫和适应性免疫 cDC的T细胞引发活性），更具体地说，如何利用这一点作为天然佐剂原理 提高流感疫苗的效力。在目标3中，Lingwood博士和Balazs博士开发了一种AAV- 基因递送平台，其在体内拯救sIL-6 R水平及其IL-6捕获活性。他们会用这个系统 通过实验确定sIL-6 R内的致病多态性如何失调IL-6捕获， 体内炎症活性，然后平行地，校正IL-6所需的治疗性sIL-6 R表达水平 体内平衡和恢复其中功能。总的来说，这项提案将定义和利用新发现的 SIL 6 R免疫防御生物学治疗人类疾病，符合NIH的目标和使命。