Study of intestinal cell differentiation with monoclonal antibodies to intestinal cell surface components.
Study of intestinal cell differentiation with monoclonal antibodies to intestinal cell surface components.
复制标题
用针对肠细胞表面成分的单克隆抗体研究肠细胞分化。
DOI：
10.1016/0012-1606(85)90482-8
复制
发表时间：
1985
影响因子：
2.7
通讯作者：
Isselbacher,KJ
中科院分区：
文献类型：
--
作者：
Quaroni,A;Isselbacher,KJ
本文报道了用不同肠细胞组分（包括成年绒毛和隐窝细胞的腔膜）和新生大鼠肠细胞免疫小鼠，将小鼠骨髓瘤（NSI）细胞与其脾细胞融合，制备了与大鼠肠绒毛和隐窝细胞质膜抗原反应的单克隆抗体。大多数抗体的抗原靶点已被确定。它们包括成年绒毛细胞刷状缘（管腔）膜的主要蛋白组分（蔗糖酶-异麦芽糖酶、麦芽糖酶、乳糖酶、氨肽酶N、碱性磷酸酶）和新鉴定的肠上皮细胞特异性蛋白抗原。在制备的25种独立衍生的单克隆抗体中，18种仅与绒毛细胞的刷状缘膜反应，证实了其独特的蛋白质组成。还获得了特异性染色隐窝细胞、存在于绒毛下半部的新分化的上皮细胞、顶部绒毛细胞以及绒毛和隐窝细胞的抗体并进行了表征。这些抗体已用于研究肠上皮分化和发育过程中细胞和组织特异性功能的表达。与用多克隆抗血清获得的结果相反，在隐窝细胞中未检测到无活性形式的刷状缘酶。在不同水平的绒毛，并在未分化和分化的肠细胞中表达的细胞表面成分的鉴定表明，在肠上皮细胞的分化是一个连续的和渐进的过程，涉及转录和翻译调控的不同的基因集。
Monoclonal antibodies that react with antigens of the plasma membrane of rat intestinal villus and crypt cells have been prepared by fusion of mouse myeloma (NSI) cells with spleen cells of mice immunized with various intestinal cellular fractions, including the luminal membrane of adult villus and crypt cells, and of newborn rat intestinal cells. The antigenic targets of most antibodies have been identified. They include major protein components of the brush border (luminal) membrane of adult villus cells (sucrase-isomaltase, maltase, lactase, aminopeptidase N, alkaline phosphatase) and newly identified protein antigens specific for intestinal epithelial cells. Of 25 independently derived monoclonal antibodies prepared, 18 reacted exclusively with the brush border membrane of the villus cells, confirming its unique protein composition. Antibodies specifically staining the crypt cells, the newly differentiated epithelial cells present in the lower half of the villi, the top villus cells, and both villus and crypt cells were also obtained and characterized. These antibodies have been used to study the expression of cell- and tissue-specific functions during differentiation and development of the intestinal epithelium. Contrary to results obtained with polyclonal antisera, no inactive forms of the brush border enzymes have been detected in the crypt cells. The identification of cell surface components expressed at different levels of the villi, and in both undifferentiated and differentiated intestinal cells, suggests that cell differentiation in the intestinal epithelium is a continuous and gradual process involving both transcriptional and translational regulation of different sets of genes.