Mucosal Immunity of gaitro-intestinal tract-making in vitro modol of GI tract

上消化道的粘膜免疫制作胃肠道体外模型

• 批准号: 10670732
• 负责人: ODA Megumi
• 金额: $ 2.11万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670732/
• 关键词: Immune organ; gastro intestinal tract; muccsal immunity; chemokine; mast cell; in vitro model

Background and objective : To clarify a role of the gastrointestinal tract in mediating immunity, we examined following three points between 1998 and 1999, using experiments where gastrointestinal epithelial cells are infected by bacteria and viruses.(1) Whether mRNA of the chemokine family is expressed in epithelial cells, (2) whether the chemokine mRNAs are translated into proteins and (3) whether the chemokine proteins have any functions. As a result, the patterns of chemokines produced by gastrointestinal epithelial cells due to bacterial or viral infections differed from each other, which was thought to be the main cause of the diversity in local mobilization of inflammatory cells and inflammatory responses against various infections. Furthermore, IL-8 obtained from supernatant of cell culture was concentrated, then added to neutrophils and briefly cultured. Subsequently, expression levels of CD11b/18 and Leu8 (L-selectin) on the surface of neutrophils were measured by FACS, resul … More ting in an increase in CD11b/18 and a decrease in L-selectin expression. And anti-IL-8 antibody partially inhibited these increases and decreases, suggesting that IL-8 in supernatant of cell culture plays a role in activating neutrophils. However, other cytokines are also involved.Then in 2000, we used an Ussing chamber to electrophysiologically analyze the influence of various chemokines and supernatant of cell culture on T84 and/or 18CO (human gastrointestinal fibroblasts) and/or neutrophils coculture systems. In addition, establishment of a mast cell line was attempted, to examine the effects of mast cells instead of neutrophils.Methods and Results : Coculture of gastrointestinal epithelial cells with fibroblasts facilitated membrane depolarization, and coculture with neutrophils further facilitated depolarization. A mast cell line was established using a culture system where IL-6, stem cell factor and prostaglandin E_2 were added to mononuclear cells from cord blood. We intend to further investigate the coculture system with this cell line.The pathogenesis and the treatment strategy of 0-157 infection were also investigated. Less

背景和目的：为了阐明胃肠道在介导免疫中的作用，我们在1998年至1999年期间，使用细菌和病毒感染胃肠道上皮细胞的实验，研究了以下三点。(1)趋化因子家族的mRNA是否在上皮细胞中表达，（2）趋化因子mRNA是否翻译成蛋白质，以及（3）趋化因子蛋白质是否具有任何功能。因此，由于细菌或病毒感染，胃肠上皮细胞产生的趋化因子的模式彼此不同，这被认为是炎症细胞局部动员和炎症反应对各种感染的多样性的主要原因。此外，将从细胞培养物的上清液中获得的IL-8浓缩，然后添加到嗜中性粒细胞中并短暂培养。随后，通过流式细胞仪检测中性粒细胞表面CD 11b/18和Leu 8（L-选择素）的表达水平，结果显示， ...更多信息 CD 11b/18表达增加，L-选择素表达减少。抗IL-8抗体部分抑制了这些增加和减少，表明细胞培养上清中的IL-8在激活中性粒细胞中发挥作用。2000年，我们利用Ussing小室电生理分析了各种趋化因子和细胞培养上清对T84和/或18 CO（人胃肠道成纤维细胞）和/或中性粒细胞共培养系统的影响。此外，建立一个肥大细胞系的尝试，检查肥大细胞的影响，而不是neutrophils.Methods和结果：共培养的胃肠道上皮细胞与成纤维细胞促进膜去极化，与中性粒细胞共培养进一步促进去极化。用脐血单个核细胞加入IL-6、干细胞因子和前列腺素E_2建立肥大细胞系。本研究拟进一步探讨与该细胞系的共培养体系，以及0-157感染的发病机制和治疗策略。少

项目成果

N.Yamashita,M.Oda et al: "Molecular Detection of Metastatic Retinoblastoma Cells by Transcription Polymerase Reaction for Interphotoreceptor Retinoid-binding protein mRNA"Cancer. (in press). (2001)

N.Yamashita、M.Oda 等人：“通过转录聚合酶反应对光感受器间视黄醇结合蛋白 mRNA 进行转移性视网膜母细胞瘤细胞的分子检测”癌症。

小田慈: "病原性大腸菌O157患者の臨床医学的解析"病原性大腸菌O157による疾患の重症化及び治療を目的とした医薬品の開発研究 研究報告書(研究代表者 竹田多恵). 128-129 (2000)

小田惠：“致病性大肠杆菌O157患者的临床医学分析”旨在加重和治疗致病性大肠杆菌O157引起的疾病的药物的开发和研究研究报告（首席研究员武田泰江）128-129（2000年）。

井上拓也,小田慈 他: "腸管出血性大腸菌O-157感染による溶血性尿毒症症候群に胆汁鬱滞反復性尿道感染を合併し、重篤な肝障害をきたした一例"小児感染免疫. (in press). (2001)

Takuy​​a Inoue、Megumi Oda 等：“肠出血性大肠杆菌 O-157 感染合并胆汁淤积和反复尿道感染，导致严重肝损伤导致溶血性尿毒症综合征一例”（《儿科感染免疫学》）（2001 年出版）。 ）

Megumi Oda et al.: "Treatment and pathogenesis of Nemclytic Unmic syndrme caused by E.cili c-157 Annual Review, Blood 1998"Igukushoin, Tokyo. 56-63 (1998)

Megumi Oda 等人：“E.cili c-157 年度回顾，血液 1998 年引起的 Nemclytic Unmic 综合征的治疗和发病机制”Igukushoin，东京。

Megumi Oda: "Clinical evaruation of the patients infected by E.Coli 0157"Research Report 42282 (Head Investigator T.Tae). 128-129 (2000)

Megumi Oda：“大肠杆菌 0157 感染患者的临床评估”研究报告 42282（首席研究员 T.Tae）。