COCULTURE OF EPITHELIAL CELLS AND MUCOSAL LYMPHOCYTES

肠粘膜环境包含多种细胞类型， 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。先前的体外 粘膜生理学和免疫学的模型依赖于 上皮细胞或白细胞。本提案的目的是 探讨粘膜淋巴细胞与上皮细胞共培养的作用 细胞在生理学、生物化学和分子生物学水平上。 我的赞助商的实验室已经开发了共培养系统。 在这个建议中，实验设计来研究生产 细胞因子对共培养的反应，以及它们对上皮细胞的作用。 单层差异显示聚合酶链反应也将被用于 用于鉴定其转录被新调节的基因， 对共同文化的回应。最后，一组单克隆抗体， 将筛选共培养系统，以鉴别 表达的表位及其与粘膜生理学的相关性。 预计这些实验将有助于加深对 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， 这项提案将检验上皮内淋巴细胞充当 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.