Role of Gender-associated Microbiota in Organ-specific Autoimmunity

性别相关微生物群在器官特异性自身免疫中的作用

• 批准号: 8465822
• 负责人: Martin A. Kriegel
• 金额: $ 13.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8465822
• 关键词: Age; Ampicillin; Antibiotic Therapy; Antibiotics; Autoimmune Diseases; Autoimmune Responses; Autoimmunity; Bacteria; Bacteroides; Bifidobacterium; CCR9 gene; Cecum; Cell physiology; Cells; Colon; Combined Antibiotics; DNA; DNA amplification; Data; Development; Diabetes Mellitus; Disease; Distal; Equilibrium; Feces; Female; Flow Cytometry; Future; Gender; Gender Role; Gene Expression Profiling; Genomics; Germ-Free; Goals; Gonadal Steroid Hormones; Homing; Hormonal; Housing; Human; Immune; Immune system; Immunologic Markers; Inbred NOD Mice; Incidence; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Interleukin-2; Intestines; Lactobacillus; Lamina Propria; Lead; Life; Lymphocyte; Mediating; Metronidazole; Modeling; Monitor; Mono-S; Mouse Strains; Mus; Neomycin; Oral; Organ; Parents; Pathogenesis; Penetrance; Phenotype; Population; RNA; Regimen; Regulatory T-Lymphocyte; Research; Ribosomal DNA; Running; Sampling; Sex Bias; Sex Characteristics; Small Intestines; Sorting - Cell Movement; Specific Pathogen Frees; T-Lymphocyte; T-Lymphocyte Subsets; Taxon; Tetracyclines; Time; Tissues; Transplantation; Vancomycin; Weaning; base; cohort; commensal microbes; gastrointestinal; germ free condition; gut microbiota; insight; integrin beta7; male; microbiome; mouse model; next generation; novel; novel strategies; prevent; protective effect; pyrosequencing; receptor; research study; treatment strategy; young adult

DESCRIPTION (provided by applicant): Type 1 diabetes is an organ-specific autoimmune disease that requires life-long monitoring and treatment to prevent damage of multiple organs. The research proposed herein aims to characterize protective commensal bacteria influencing the pathogenesis of type 1 diabetes in a prototypical murine model, the non-obese diabetic (NOD) mouse. The hypothesis is based on the fact that the incidence of type 1 diabetes is markedly lower in male than female NOD mice under specific-pathogen-free (SPF) conditions that allow colonization with commensals. However, the absence of microbiota under germ-free conditions leads to 100% penetrance of the phenotype in both genders. It is hypothesized that gut microbiota mediate this sex bias by influencing the spontaneous autoimmune response. We therefore plan to analyze potential candidate microbiota as well as the microbiome in male, female and sex-hormone-ablated mice. We will also dissect the immunological differences between both genders, with a focus on T cell subsets, given their importance in the pathogenesis of this disease. Finally, we plan to study the influence of oral gavage with candidate microbiota and antibiotic treatments on disease development and T cell function. We specifically plan to achieve these goals as follows: Specific Aim 1: To examine candidate microbiota and the unbiased microbiome in male, female and sex-hormone-ablated NOD mice. We will perform next-generation pyrosequencing of each gastrointestinal segment from NOD mice. We will analyze the microbiome and selected candidate microbiota for possible gender differences in young adult NOD mice and at multiple time points from weaning age until development of disease. Specific Aim 2: To analysis the T cell compartment in the small intestine and colon of male versus female NOD mice. We will analyze various T cell subsets using flow cytometry and RNA microarray profiling to identify functional differences between male and female gastrointestinal immune compartments. Specific Aim 3: To gavage SPF-housed female and germ-free male NOD mice with potentially protective microbiota that have been identified in Specific Aim 1, and to perturb the host with selective antibiotic combinations. We will also investigate any alterations in T cell functions induced by these manipulations. We anticipate that the proposed research will lead to discovery of new mechanisms in the pathogenesis of type 1 diabetes, and opens the novel avenue of gender-associated microbiota in autoimmunity in general. The research is expected to have broad implications for sex-biased autoimmune diseases with the potential for development of new treatment strategies. The project deals with the influence of gender-associated commensal gut bacteria on type 1 diabetes in the non-obese diabetic (NOD) mouse strain. We plan to identify beneficial gut bacteria that protect male NOD mice under standard housing conditions and characterize their impact on disease in highly susceptible germ-free NOD mice. Exploration of the commensals' effects on the immune system and sex bias in this mouse model will help to better understand the female preponderance of human autoimmune diseases in general, and open possibly new treatment strategies.

描述（由申请人提供）：1型糖尿病是一种器官特异性自身免疫性疾病，需要终生监测和治疗以防止多个器官受损。本文提出的研究旨在表征在典型小鼠模型（非肥胖糖尿病（NOD）小鼠）中影响 1 型糖尿病发病机制的保护性共生细菌。该假设基于以下事实：在允许共生菌定植的无特定病原体 (SPF) 条件下，雄性 NOD 小鼠的 1 型糖尿病发病率明显低于雌性 NOD 小鼠。然而，在无菌条件下缺乏微生物群导致两种性别的表型外显率均为 100%。据推测，肠道微生物群通过影响自发的自身免疫反应来介导这种性别偏见。因此，我们计划分析潜在的候选微生物群以及雄性、雌性和性激素消除小鼠的微生物组。我们还将剖析两性之间的免疫学差异，重点关注 T 细胞亚群，因为它们在这种疾病发病机制中的重要性。最后，我们计划研究口服强饲候选微生物群和抗生素治疗对疾病发展和 T 细胞功能的影响。我们具体计划实现以下目标： 具体目标 1：检查雄性、雌性和性激素消除 NOD 小鼠的候选微生物群和无偏微生物组。我们将对 NOD 小鼠的每个胃肠节段进行下一代焦磷酸测序。我们将分析年轻成年 NOD 小鼠以及从断奶年龄到疾病发生的多个时间点的微生物组和选定的候选微生物群可能存在的性别差异。具体目标 2：分析雄性和雌性 NOD 小鼠小肠和结肠中的 T 细胞区室。我们将使用流式细胞术和 RNA 微阵列分析来分析各种 T 细胞亚群，以确定男性和女性胃肠道免疫区室之间的功能差异。具体目标 3：用特定目标 1 中确定的具有潜在保护性微生物群的 SPF 饲养雌性和无菌雄性 NOD 小鼠进行灌胃，并用选择性抗生素组合扰乱宿主。我们还将研究这些操作引起的 T 细胞功能的任何改变。我们预计，拟议的研究将导致 1 型糖尿病发病机制的新机制的发现，并为自身免疫中性别相关微生物群的研究开辟新途径。该研究预计将对性别偏见的自身免疫性疾病产生广泛影响，并有可能开发新的治疗策略。该项目研究了性别相关的共生肠道细菌对非肥胖糖尿病 (NOD) 小鼠品系 1 型糖尿病的影响。我们计划鉴定在标准饲养条件下保护雄性 NOD 小鼠的有益肠道细菌，并表征它们对高度易感的无菌 NOD 小鼠疾病的影响。在该小鼠模型中探索共生体对免疫系统的影响和性别偏见将有助于更好地了解女性在人类自身免疫性疾病中的总体优势，并开辟可能的新治疗策略。