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

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年期间通过胃肠道上皮细胞被细菌和病毒感染的实验，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是否被翻译成蛋白质；(3)趋化因子蛋白是否具有任何功能。因此，由于细菌或病毒感染，胃肠道上皮细胞产生的趋化因子的模式不同，这被认为是炎症细胞局部动员和针对各种感染的炎症反应多样性的主要原因。将细胞培养上清中提取的IL-8浓缩后加入中性粒细胞中短暂培养。随后，用流式细胞仪检测中性粒细胞表面CD11b/18和l -选择素（Leu8）的表达水平，结果显示CD11b/18表达升高，l -选择素表达降低。而抗IL-8抗体部分抑制了这种增减，提示细胞培养上清中的IL-8具有活化中性粒细胞的作用。然而，其他细胞因子也参与其中。然后在2000年，我们使用Ussing室电生理学分析了各种趋化因子和细胞培养上清对T84和/或18CO（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，还尝试建立肥大细胞系，以检验肥大细胞而非中性粒细胞的作用。方法与结果：胃肠道上皮细胞与成纤维细胞共培养可促进膜去极化，与中性粒细胞共培养可进一步促进去极化。将IL-6、干细胞因子和前列腺素E_2添加到脐带血单核细胞中，建立了肥大细胞系。我们打算进一步研究该细胞系的共培养系统。并对0-157感染的发病机制和治疗策略进行了探讨。少

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