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.

项目摘要 /摘要 这是Daniel Lingwood博士（PI）和Alejandro Balazs（Co-I）的申请， 哈佛医学院和MGH，麻省理工学院和哈佛大学Ragon研究所的教职员工。他们的 实验室专门定义抗疫苗抗体反应的免疫学（lingwood）和 通过AAV介导的基因输送（BALAZS）进行免疫的工程。林伍德博士发现 抗体对疫苗接种和促炎性免疫反应的反应通常受到中央调节 通过常规树突状细胞（CDC）提供的新型控制轴，这是在小鼠中定义的单个淋巴谱系 通过转录因子ZBTBT46的表达。 Lingwood博士发现CDC负责IN IL-6受体（SIL6R）的可溶性，循环形式的体内生物发生，该形式通常从中捕获IL-6 解决方案以系统地协调其信号活性。 IL-6是释放的有效促炎细胞因子 在免疫挑战或组织损伤之后，SIL6R捕获的失调导致 高生理IL-6水平和炎症性疾病。该提议将检验一个中心假设，即 CDC对SIL6R的生物发生形成了一个集中的控制轴，可以调节以增强免疫力 疾病状态下的防御和恢复IL-6同种异体。在AIM 1中，Lingwood博士将定义 金属蛋白酶将ADAM10和ADAM17从CDC细胞表面释放到全身SIL6R。这些 蛋白酶将表面IL6R切割至从组织培养细胞系的Sil6r裂解，从而增强了可吸毒的靶标 调节IL-6免疫作用。但是，ADAM10/17淘汰是胚胎致死的，阻止了机械 评估其在体内活动。 Lingwood博士现在已选择性地指导ADAM10和/或ADAM17 敲除CDC谱系的敲除，并将使用这些转基因动物在两个CDC中定义ADAM活性 依赖参数：在免疫挑战后循环SIL6R设定点和SIL6R生物发生的诱导。 Lingwood博士还发现，CDC衍生的SIL6R以及随后通过 SIL6R：IL-6复合物，对于B细胞分化与抗体分泌浆细胞的必要条件是必要的 用多种蛋白质疫苗抗原免疫后的抗体输出。在AIM 2中，Lingwood博士将 定义CDC衍生的SIL6R如何通过此途径链接先天性和适应性免疫（在传统之外 CDC的T细胞启动活性），更具体地说是如何将其作为天然佐剂原理利用 提高流感疫苗的功效。在AIM 3中，Lingwood博士和Balazs博士开发了一个AAV- 拯救SIL6R水平及其IL-6捕获活动的基因输送平台。他们将使用这个系统 通过实验定义SIL6R内部的致病多态性如何使IL-6捕获和 体内炎症活性，然后并联，校正IL-6所需的治疗SIL6R表达水平 稳态和还原功能。总的来说，该建议将定义和利用新发现的 SIL6R免疫防御的生物学治疗人类疾病，与NIH的目标和使命一致。