INTESTINAL CELL DIFFERENTIATION IN VIVO AND IN CULTURE

其目的是确定的因素和细胞 诱导和调节成熟和分化的相互作用 肠上皮 本提案中概述的研究将 专注于肠细胞成分，由先前产生的 单克隆抗体由于其细胞和/或组织 在成虫肠道和不同发育阶段的分布情况 可能参与这些过程。 新的单克隆抗体将 制备并表征以进一步确定 肠隐窝细胞，并在早期检测单个细胞类型， 承诺或分化的过程。 其组成、结构和 新鉴定的抗原的生物合成将通过 结合体内和培养中进行的研究，使用肠 上皮细胞系和胎鼠肠器官培养物。 的 由不同单克隆抗体识别的抗原将被纯化 通过亲和层析并通过平板凝胶电泳表征， 免疫印迹，肽图谱，氨基酸序列测定， 纯化的肠mRNA的无细胞翻译和cDNA克隆。 的 在不同的温度下，各种抗原在肠粘膜中的分布 发展和分化的阶段，并在培养的细胞，将是 通过免疫荧光染色和 免疫电镜 识别不同隐窝细胞的抗体 类型将用于超微结构的比较研究，相对 在隐窝中的位置、增殖活性和胎儿中的发育， 新生大鼠的细胞，他们识别。 这些研究将提供新的 了解肠细胞分化的过程， 有助于我们了解疾病状态下的肠粘膜， 如口炎性腹泻、炎症性肠病、与管腔成分的相互作用， 和肠道适应。

The objective is the identification of the factors and cellular interactions which induce and modulate maturation and differentiation of the intestinal epithelium. The studies outlined in this proposal will focus on intestinal cell components defined by previously produced monoclonal antibodies which, because of their cell and/or tissue distribution in adult intestine and at different stages of development are potentially involved in those processes. New monoclonal antibodies will be prepared and characterized to further define the biological properties of the intestinal crypt cells and to detect individual cell types early in the process of commitment or differentiation. The composition, structure and biosynthesis of the newly identified antigens will be investigated by a combination of studies conducted in vivo and in culture, using intestinal epithelial cell lines and fetal rat intestinal organ cultures. The antigens recognized by the different monoclonal antibodies will be purified by affinity chromatography and characterized by slab gel electrophoresis, immunoblotting, peptide mapping, amino acid sequence determination, cell-free translation of purified intestinal mRNA, and cDNA cloning. The distribution of the various antigens in the intestinal mucosa at different stages of development and differentiation, and in cultured cells, will be determined by immunofluorescence staining and by immunoelectron-microscopy. Antibodies recognizing distinct crypt cell types will be used for a comparative study of the ultrastructure, relative position in the crypts, proliferative activity and development in fetal and newborn rats of the cells they identify. The studies should provide new insights into the process of intestinal cell differentiation, and also contribute to our knowledge of the intestinal mucosa in disease states such as sprue, inflammatory bowel diseases, interaction with luminal components, and intestinal adaptation.