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: 10093221
• 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/10093221
• 关键词: Adjuvant; Animals; Antibodies; Antibody Response; Antigens; B cell differentiation; B-Lymphocytes; Biogenesis; Biology; Blood Circulation; Buffers; Cell Line; Cell Lineage; Cell membrane; Cell surface; Cells; Cleaved cell; Complex; Data; Dendritic Cells; Disease; Drug Targeting; 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; Institutes; 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; 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

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.

项目概要/摘要 这是由 Daniel Lingwood 博士 (PI) 和 Alejandro Balazs 博士 (Co-I) 助理教授提出的申请 哈佛医学院以及麻省总医院、麻省理工学院和哈佛大学拉贡研究所的教职人员。他们的 实验室专门研究疫苗抗体反应的免疫学 (Lingwood) 和 通过 AAV 介导的基因传递进行免疫工程 (Balazs)。林伍德博士发现 对疫苗接种的抗体反应以及一般的促炎性免疫反应均受到集中调节 由传统树突状细胞（CDC）提供的新型控制轴，这是小鼠中定义的单一淋巴谱系 通过转录因子 Zbtbt46 的表达。 Lingwood 博士发现，cDC 是造成这种情况的罪魁祸首。 IL-6 受体 (sIL6R) 可溶性循环形式的体内生物合成，通常从细胞中捕获 IL-6 系统地协调其信号活动的解决方案。 IL-6 是一种有效的促炎细胞因子 免疫挑战或组织损伤后，sIL6R 捕获的失调会导致 超生理性 IL-6 水平和炎症性疾病。该提案将检验中心假设： CDC 的 sIL6R 生物发生形成了一个集中控制轴，可以对其进行调节以增强免疫 防御并恢复疾病状态下的 IL-6 稳态。在目标 1 中，Lingwood 博士将定义以下贡献： 金属蛋白酶 ADAM10 和 ADAM17 影响 cDC 细胞表面系统性 sIL6R 的释放。这些 蛋白酶裂解表面 IL6R，从组织培养细胞系中释放 sIL6R，从而增强可药物靶标 调节 IL-6 免疫作用。然而，ADAM10/17 敲除具有胚胎致死性，可防止机械性 评估它们的体内活性。 Lingwood 博士现已选择性地指导 ADAM10 和/或 ADAM17 敲除 CDC 谱系，并将使用这些转基因动物来定义两个 CDC 内的 ADAM 活性 依赖参数：循环 sIL6R 设定点和免疫攻击后 sIL6R 生物发生的诱导。 Lingwood 博士还发现了 cDC 衍生的 sIL6R，以及随后通过 sIL6R：IL-6 复合物，是 B 细胞分化为分泌抗体的浆细胞所必需的，并调节 用多种蛋白质疫苗抗原免疫后产生抗体。在目标 2 中，Lingwood 博士将 定义 CDC 衍生的 sIL6R 如何通过此途径（在传统的 cDC 的 T 细胞启动活性），更具体地说，如何将其用作天然佐剂原理 提高流感疫苗的功效。在目标 3 中，Lingwood 博士和 Balazs 博士开发了一种 AAV- 拯救 sIL6R 水平及其体内 IL-6 捕获活性的基因递送平台。他们将使用这个系统 通过实验确定 sIL6R 内的致病多态性如何失调 IL-6 捕获和 体内炎症活动，然后平行地，校正 IL-6 所需的治疗性 sIL6R 表达水平 体内平衡并恢复其中的功能。总的来说，该提案将定义和利用新发现的 sIL6R 免疫防御生物学治疗人类疾病，与 NIH 的目标和使命一致。