HETEROGENEITY AND CONTACT-DEPENDENT REGULATION OF HORMONE-SECRETION BY INDIVIDUAL B-CELLS
HETEROGENEITY AND CONTACT-DEPENDENT REGULATION OF HORMONE-SECRETION BY INDIVIDUAL B-CELLS
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DOI：
10.1016/0014-4827(86)90354-x
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发表时间：
1986-02-01
影响因子：
3.7
通讯作者：
MEDA, P
中科院分区：
文献类型：
--
作者：
SALOMON, D;MEDA, P
开发了反向溶血空斑测定以可视化来自个体成人pancratic B细胞的胰岛素释放。将通过机械分散分离的大鼠胰岛获得的细胞与蛋白A包被的绵羊红细胞混合，并在已知影响胰岛素释放的条件下，在抗胰岛素血清存在下孵育。将细胞混合物与补体进一步孵育并最终固定。胰岛素释放通过溶血性斑块的存在来揭示，溶血性斑块是由补体介导的带有与蛋白A结合的胰岛素-抗胰岛素复合物的红细胞溶解引起的。台盼蓝未染色和免疫化学鉴定的B细胞周围溶血斑块的定量显示，胰岛素释放的刺激导致分泌型B细胞数量的增加以及单个B细胞反应的增强。抑制胰岛素释放时发生相反的变化。新鲜分散和一天培养的制剂的比较没有发现未受损的B细胞的分泌反应的显着差异。在这两种制剂中，单个B细胞对促分泌素的反应比例较小，程度也低于B细胞与其邻居保持或恢复接触和连接通讯的群集。然而，总体优势反应的集群是小于预期的数量，个别分泌B细胞，他们所包含的。 数据显示，B细胞在其释放胰岛素的能力方面是异质的，并提供了细胞间粘附和/或连接通讯调节个体B细胞的激素分泌的证据。
A reverse hemolytic plaque assay was developed to visualize insulin release from individual adult pancratic B cells. Cells obtained by mechanical dispersion of isolated rat islets of Langerhans were mixed with protein A-coated sheep red blood cells and incubated in the presence of an anti-insulin serum, under conditions known to affect insulin release. The cell mixture was further incubated with complement and finally fixed. Insulin release was revealed by the presence of hemolytic plaques which resulted from the complement-mediated lysis of red blood cells bearing insulin-anti-insulin complexes bound to protein A. Quantitation of hemolytic plaques around trypan blue-unstained and immunohistochemically identified B cells showed that stimulation of insulin release results in the recruitment of increasing numbers of secreting B cells as well as in the enhanced response of individual B cells. Reverse changes occur upon inhibition of insulin release. Comparison of freshly dispersed and one-day-cultured preparations did not reveal significant differences in the secretory response of undamaged B cells. In both preparations, single B cells responded to secretagogues in smaller proportions and to a lesser extent than clusters in which B cells had either maintained or restroed contacts and junctional communication with their neighbours. However, the overall preponderant response of clusters was less than expected from the number of individually secreting B cells they contained. The data show that B cells are heterogeneous in terms of their ability to release insulin and provide evidence that cell-to-cell adhesion and/or junctional communication regulate hormone secretion from individual B cells.