Stat3 Activation in Bacteria-Induced Murine Th17 Predominant Colonic Inflammation

细菌诱导的小鼠 Th17 主要结肠炎症中 Stat3 的激活

• 批准号: 7741631
• 负责人: Shervin Rabizadeh
• 金额: $ 5.89万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-03 至 2009-07-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7741631
• 关键词: Acute; Address; Adult; Age; Anaerobic Bacteria; Anesthesia procedures; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Antibodies; Apoptosis; Bacteria; Bacteroides fragilis; Binding; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Cecum; Cells; Chronic; Class; Clinical; Colitis; Colon; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Crohn' s disease; Cytokine Signaling; Data; Dependency; Diarrhea; Disease; Electrophoretic Mobility Shift Assay; Epithelial; Epithelial Cells; Epithelium; Exhibits; Family; Flow Cytometry; Generations; Genus Cola; Harvest; Hemorrhage; Human; IL17 gene; Immune; Immune response; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Interleukin-10; Interleukin-17; Interleukin-6; Intestinal Neoplasms; Intestines; Lamina Propria; Location; Lymphocyte; Mediator of activation protein; Metalloproteases; Modeling; Molecular; Monitor; Mus; Oral; Outcome; Pathogenesis; Pathology; Pathway interactions; Patients; Personal Satisfaction; Phenotype; Placebos; Property; Public Health; Rate; Recombinants; Research; Research Project Grants; Resistance; Rodent; Role; STAT protein; Signal Transduction; Spleen; Staining method; Stains; Stat3 protein; T-Lymphocyte; Therapeutic Intervention; Thinking; Time; Tissues; Toxin; Virulence Factors; Week; Weight; Work; Zinc; cell type; data modeling; day; human disease; human study; in vivo; macrophage; neutrophil; pathogen; promoter; rectal; research study; response; transcription factor; tumor; tumor initiation

DESCRIPTION (provided by applicant): Bacteroides fragilis is a common human commensal residing in the colon. B. fragilis enterotoxigenic (ETBF) class has been associated with inflammatory diarrheal disease, inflammatory bowel disease, and colorectal cancer. Recent murine model data has shown that chronic colonization with ETBF leads to a proliferative and proinflammatory response which can act as a co-factor in colon tumor initiation and promotion specifically in the multiple intestinal neoplasia (Min) model. Signal transducer and activator of transcription 3 (StatS) has both pro- and anti-inflammatory properties through activation by IL-6 and IL-10, respectively. Phosphorylated StatS is found in intestinal T cells from patients with chronic gut inflammation such as Crohn's disease most likely inhibiting apoptosis of T cells. It also is present in tumors where it functions as a tumor immune suppressor. Though important in chronic inflammation and tumors, its role has not been well studied. Specific Aim: To determine the effect of ETBF colonization on StatS in colonic tumor formation in Min mice and inflammation in a colitis-associated murine model. Mice (C57BL/6 and Min) Swks to 3-4mo of age will be used in the proposed experiments. The C57BL/6 mice will be used to demonstrate StatS activity in inflammation of the colitis-associated model while the Min mice will be used to analyze StatS activity in colonic tumor formation. After pre-treatment with antibiotics, under anesthesia, mice will be inoculated via oral gavage with ETBF, non-toxic B. fragilis, or placebo. Mice will be closely monitored in terms of clinical status (weight, activity, rectal bleeding, diarrhea) and sacrificed at various time points post inoculation (1 day to 3-4 months). Colon, cecum, and spleen will be harvested and used for pathology. Immunofluorescent staining using specific antibodies and Electophoretic mobility shift assay (EMSA) will also be used to demonstrate presence of phosphorylated StatS. Also tissue will be utilized for flow cytometry analysis of specific locations (epithelium, lamina propria) and cell types (lymphocytes, macrophages, neutrophils, etc) for StatS activity. This research will be relevant to public health in several ways. It will further define the potential role and pathway of a bacterium, ETBF, in inflammation and proliferation that could relate to disorders such as colon cancer and Inflammatory Bowel Disease. Furthermore it will establish a model in which therapeutic interventions could be attempted in hopes of treating such deadly prevalent diseases.

描述（由申请人提供）：bacteroides fragilis是居住在结肠中的常见人类共生。 B. fragilis肠毒素（ETBF）类别与炎症性腹泻病，炎症性肠病和结直肠癌有关。最近的鼠模型数据表明，具有ETBF的慢性定殖导致了增殖和促炎反应，该反应可以作为结肠肿瘤起始和促进的共同因素，特别是在多个肠道肿瘤（Min）模型中。转录3（统计）的信号换能器和激活因子分别通过IL-6和IL-10激活具有促炎和抗炎特性。在患有慢性肠道炎症患者的肠道T细胞中发现了磷酸化的数据，例如克罗恩病，最可能抑制T细胞凋亡。它也存在于肿瘤中充当肿瘤免疫抑制因子。尽管在慢性炎症和肿瘤中很重要，但其作用尚未得到很好的研究。具体目的：确定Min小鼠中ETBF定殖对结肠肿瘤形成的统计数据的影响以及与结肠炎相关的鼠模型中的炎症。在拟议的实验中，将使用小鼠（C57BL/6和min）至3-4MO的小鼠。 C57BL/6小鼠将用于证明与结肠炎相关模型炎症中的统计性活性，而最小小鼠将用于分析结肠肿瘤形成中的Stats活性。在用抗生素进行预处理后，在麻醉下，将通过ETBF，无毒的脆弱芽孢杆菌或安慰剂接种小鼠。小鼠将根据临床状况（体重，活动，直肠出血，腹泻）进行密切监测，并在接种后的各个时间点处死（1天至3-4个月）。结肠，盲肠和脾脏将被收获并用于病理学。使用特定抗体和电载迁移率转移测定法（EMSA）的免疫荧光染色也将用于证明存在磷酸化的数据。同样，将组织用于特定位置（上皮，层）和细胞类型（淋巴细胞，巨噬细胞，中性粒细胞等）的流式细胞仪分析。这项研究将以几种方式与公共卫生有关。它将进一步定义细菌ETBF在炎症和增殖中的潜在作用和途径，这可能与结肠癌和炎症性肠病等疾病有关。此外，它将建立一个模型，在该模型中，可以尝试尝试治疗干预措施，以期治疗这种致命的普遍性疾病。