Leveraging humoral immunity to promote commensal microbial protection from T1D

利用体液免疫促进共生微生物对 T1D 的保护

• 批准号: 10196996
• 负责人: Michael A Silverman
• 金额: $ 22万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-17 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10196996
• 关键词: Adaptive Immune System; Affect; Alleles; Antibodies; Antibody Response; Antigen Presentation; Antigens; Autoantigens; Autoimmune Diabetes; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; CD4 Positive T Lymphocytes; Cell Compartmentation; Cells; Child; Development; Diabetes Mellitus; Disease; Dominant Genetic Conditions; Enhancers; Environmental Risk Factor; Exposure to; FOXP3 gene; Flow Cytometry; Future; Generations; Genetic; Genetic Models; Goals; HLA Antigens; Haplotypes; Histocompatibility Antigens Class II; Human; Humoral Immunities; IgG1; Immune system; Immunoglobulin A; Immunologist; Inbred NOD Mice; Incidence; Insulin; Insulin-Dependent Diabetes Mellitus; Interdisciplinary Study; Intestines; Investigation; Lead; Life; Major Histocompatibility Complex; Methodology; Methods; Microbe; Modeling; Molecular; Mus; Pancreas; Pathway interactions; Patients; Pennsylvania; Peripheral; Planets; Population; Regulatory T-Lymphocyte; Reporting; Risk; Role; Sampling; Shapes; System; Systems Development; T-Lymphocyte; Techniques; Testing; Transgenic Organisms; Universities; Work; autoreactive T cell; base; commensal microbes; early life exposure; effective therapy; genetic linkage; gut microbiome; gut microbiota; human model; immunoregulation; innovation; insight; insulitis; microbial; microbiome; microbiome analysis; microbiota; microorganism interaction; mouse model; natural antibodies; prevent; tool

PROJECT SUMMARY Type 1 diabetes (T1D) is an autoimmune disease that affects millions of people worldwide. The incidence of T1D is rising, especially in young children. Although significant progress has been made to predict who is at risk for developing T1D, there are no effective therapies to prevent this disease. Both genetic and environmental factors contribute to the risk of developing T1D. Certain human leukocyte antigen (HLA) haplotypes dominantly protect against the development of T1D, yet the mechanism of this remarkable protection from autoimmunity is not well-understood. NOD mice, the most widely used model of T1D, do not express a major histocompatibility complex (MHC) class II E molecule. Transgenic expression of the MHCII E molecule in NOD mice (Eα16/NOD) completely prevents T1D, mirroring dominant HLA protection from T1D in humans. Using these Eα16/NOD mice as a model of dominant genetic protection from T1D, we recently demonstrated that MHC/HLA genetic protection from autoimmunity operates via immune system selection of diabetes-protective commensal microbiota early in life, which in turn, influences the developing immune system. Since we find an increased proportion of intestinal Tregs and a distinct early-life microbiome in Ea16/NOD mice, our central hypothesis is that specific microbes prevent T1D by promoting development of diabetes-protective CD4+ regulatory T cells (Tregs). Modeling of HLA class II dominant protection from T1D in murine models may provide critical insights to support our long-term goal of developing microbiota-based therapies to prevent T1D in humans. Due to the complexity and high levels of variability of the intestinal microbiome, determining the specific microbial strains that educate the immune system is problematic. Building upon established techniques of microbial flow cytometry of antibody-coated commensal microbes, we developed an innovative approach called mFLOW-SEQ to identify microbes that stimulate the CD4+ T cell compartment in a model of genetic protection from T1D. Aim 1 leverages our innovative mFLOW- SEQ approach to identify commensal microbes that induce CD4+ Treg cells and prevent T1D. Aim 2 uses genetic models to determine the extent to which early-life Treg cells prevent autoimmunity in genetically protected Eα16/NOD mice. Successful completion of these aims will provide critical information on which early- life microbes impact the development of the immune system to prevent T1D and further whether early-life Tregs provide protection from T1D. In addition, mFLOW-SEQ is an innovative tool for identifying microbes that modulate the immune system in mice. As a future direction, we envision applying mFLOW-SEQ to human samples to help generate personalized microbiota therapies based on HLA haplotypes for human patients at risk for T1D.

项目摘要 1型糖尿病（T1 D）是一种自身免疫性疾病，影响全球数百万人。的发生率 T1 D正在上升，特别是在幼儿中。尽管在预测谁在 由于T1 D的发病风险很高，目前还没有有效的治疗方法来预防这种疾病。遗传和 环境因素会增加患T1 D的风险。人类白细胞抗原（HLA） 单倍型主要防止T1 D的发展，但这种显着的机制 对自身免疫性的保护还没有很好的理解。NOD小鼠是最广泛使用的T1 D模型， 表达主要组织相容性复合体（MHC）II类E分子。MHCII E的转基因表达 NOD小鼠（Eα16/NOD）中的HLA分子完全预防T1 D，反映了NOD小鼠中T1 D的显性HLA保护作用。 人类利用这些Eα16/NOD小鼠作为显性遗传保护T1 D的模型，我们最近 表明MHC/HLA遗传保护通过免疫系统选择自身免疫起作用， 糖尿病-生命早期的保护性肠道微生物群，这反过来又会影响发育中的免疫系统， 系统由于我们发现肠道细菌的比例增加和不同的早期微生物组， Ea 16/NOD小鼠，我们的中心假设是特定微生物通过促进T1 D来预防T1 D 糖尿病保护性CD 4+调节性T细胞（Tcells）的发育。HLA Class II优势模型 在小鼠模型中保护T1 D可能提供关键的见解，以支持我们的长期目标， 基于微生物的疗法来预防人类的T1 D。由于复杂性和高度的可变性， 肠道微生物组，确定教育免疫系统的特定微生物菌株， 有问题基于已建立的抗体包被微生物流式细胞术技术 微生物，我们开发了一种称为mFLOW-SEQ的创新方法，以识别刺激细胞生长的微生物。 T1 D遗传保护模型中的CD 4 + T细胞区室。Aim 1利用我们的创新mFLOW- SEQ方法鉴定诱导CD 4 + Treg细胞并预防T1 D的肠道微生物。Aim 2用途 遗传模型，以确定早期Treg细胞在遗传上阻止自身免疫的程度， 保护Eα16/NOD小鼠。这些目标的成功完成将提供关键信息， 生命微生物影响免疫系统的发展，以防止T1 D，并进一步是否早期生命T1 D 保护T1 D。此外，mFLOW-SEQ是一种用于鉴定微生物的创新工具， 调节小鼠的免疫系统。作为未来的发展方向，我们设想将mFLOW-SEQ应用于人类 样本，以帮助基于人类患者的HLA单倍型产生个性化的微生物群疗法， T1 D风险

项目成果

Insight Into Host-Microbe Interactions Using Microbial Flow Cytometry Coupled to Next-Generation Sequencing.

使用微生物流式细胞术与下一代测序相结合深入了解宿主-微生物相互作用。

• DOI: 10.1093/jpids/piab092
• 发表时间: 2021
• 期刊: Journal of the Pediatric Infectious Diseases Society
• 影响因子: 3.2
• 作者: Silverman,MichaelA;Green,JamalL
• 通讯作者: Green,JamalL