COCULTURE OF EPITHELIAL CELLS AND MUCOSAL LYMPHOCYTES

上皮细胞和粘膜淋巴细胞的培养

• 批准号: 2713328
• 负责人: SUNIL K SHAW
• 金额: $ 0.84万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2713328
• 关键词: cell cell interaction; cytokine; gastrointestinal epithelium; intestinal mucosa; lymphocyte; mixed tissue /cell culture; monoclonal antibody; polymerase chain reaction

The intestinal mucosal milieu comprises a variety of cell types and molecules including epithelial cells, leukocytes, stromal cells, endothelial cells, a neural network, extracellular matrix, cytokines growth factors and other molecules and cell types. Previous in vitro models of mucosal physiology and immunology have relied on monocultures of epithelial cells or leukocytes. It is the aim of this proposal to investigate the effect of co-culture of mucosal lymphocytes and epithelial cells at the levels of physiology, biochemistry, and molecular biology. Co-culture systems have already been developed in my sponsor's laboratory. In this proposal, experiments are designed to investigate the production of cytokines in response to co-culture, and their effect on the epithelial monolayer. The differential display polymerase chain reaction will also be utilized to identify genes whose transcription is newly modulated in response to co-culture. Lastly, a panel of monoclonal antibodies to such a co-culture system will be screened in order to identify differentially expressed epitopes, and their relevance to mucosal physiology. These experiments are anticipated to lead to enhanced understanding of the interaction between epithelial cells and intra-epithelial lymphocytes, and its contribution to inflammatory processes in the intestine. Thus, this proposal will test the hypothesis that intra-epithelial lymphocytes serve as an important regulator of intestinal epithelial cell function.

肠粘膜环境包含多种细胞类型 分子包括上皮细胞、白细胞、基质细胞、 内皮细胞、神经网络、细胞外基质、细胞因子 生长因子和其他分子和细胞类型。以前的体外实验 粘膜生理学和免疫学模型依赖于单一培养 上皮细胞或白细胞。该提案的目的是 研究粘膜淋巴细胞与上皮细胞共培养的效果 生理学、生物化学和分子生物学水平的细胞。 共培养系统已经在我的资助者的实验室中开发出来。 在本提案中，实验旨在研究生产 细胞因子对共培养的反应及其对上皮细胞的影响 单层。差异显示聚合酶链反应也将 用于识别其转录在新调控中受到调节的基因 对共文化的反应。最后，一组针对此类的单克隆抗体 将筛选共培养系统，以区别识别 表达的表位及其与粘膜生理学的相关性。 这些实验预计将加深对 上皮细胞和上皮内淋巴细胞之间的相互作用，以及 它对肠道炎症过程的贡献。因此，这个 该提案将检验上皮内淋巴细胞发挥作用的假设 作为肠上皮细胞功能的重要调节因子。