INTESTINAL CELL DIFFERENTIATION IN VIVO AND IN CULTURE

目标是确定因素和细胞 诱导和调节细胞成熟和分化的相互作用 肠上皮细胞。这项提案中概述的研究将 重点研究以前生产的肠道细胞成分 单抗，因为它们的细胞和/或组织 在成虫肠道和不同发育阶段的分布 可能参与到这些过程中。新的单抗将是 制备和表征以进一步确定其生物学特性 肠道隐窝细胞，并在早期检测单个细胞类型 承诺或区分的过程。它的组成、结构和 新确认的抗原的生物合成将通过一项 体内和培养相结合的研究，使用肠道 上皮细胞系和胎鼠肠道器官培养。这个 不同的单抗识别的抗原将被提纯 通过亲和层析和平板凝胶电泳法进行表征， 免疫印迹、肽图谱、氨基酸序列测定、 纯化的肠道信使核糖核酸的无细胞翻译和cdna克隆。这个 不同抗原在不同时间段肠粘膜中的分布 在培养细胞中，发育和分化的阶段将是 通过免疫荧光染色和通过 免疫电子显微镜。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 胎儿和胎儿在隐窝的位置、增殖活性和发育 它们所识别的细胞的新生大鼠。这些研究应该提供新的 对肠道细胞分化过程的洞察，以及 有助于我们对疾病状态下的肠道粘膜的了解 如流行性出血热、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.