COCULTURE OF EPITHELIAL CELLS AND MUCOSAL LYMPHOCYTES

肠粘膜环境由多种细胞类型和 分子包括上皮细胞、白细胞、间质细胞、 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。先前的体外试验 粘膜生理学和免疫学的模型依赖于单一的 上皮细胞或白细胞。这项提议的目的是 黏膜淋巴细胞与上皮细胞共培养的实验研究 生理、生物化学和分子生物学水平上的细胞。 共培养系统已经在我的赞助商的实验室里开发出来了。 在这项建议中，设计了实验来调查生产 细胞因子对共培养的反应及其对上皮细胞的影响 单层。差异显示聚合酶链式反应也将 用来识别其转录受到新调控的基因 对共文化的反应。最后，一组针对这种情况的单抗 将筛选共培养系统，以区分不同的 表达的表位及其与粘膜生理学的相关性。 这些实验有望使人们更好地理解 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，这一点 该提案将检验上皮内淋巴细胞服务于 作为肠上皮细胞功能的重要调节因子。

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.