Immunodissection: use of monoclonal antibodies to isolate specific types of renal cells.
Immunodissection: use of monoclonal antibodies to isolate specific types of renal cells.
复制标题
免疫解剖：使用单克隆抗体分离特定类型的肾细胞。
DOI：
10.1152/ajprenal.1985.248.1.f1
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Garcia-Perez,A
中科院分区：
文献类型：
--
作者：
Smith,WL;Garcia-Perez,A
几个实验室已经描述了针对不同肾细胞类型特有的细胞表面抗原的抗体。可以合理地假设，每种不同类型的肾细胞具有一个或多个独特的抗原决定簇。原则上，应该可以制备针对这些细胞特异性抗原的单克隆抗体，并使用抗体作为免疫亲和试剂来分离特定肾细胞类型的群体。我们采用这种方法在一个例子中，分离犬皮质集合管（CCT）细胞。从5 g犬肾皮质可获得约10（7）个犬CCT细胞。这与通过显微切割可以合理获得的10（3）至10（4）个CCT细胞的估计值相比。相对大量的细胞的可用性使得有可能更详细地研究细胞生理学和微管特异性过程的生物化学，如溶质和水的运输和激素作用。我们尝试分离犬CCT细胞和其他肾上皮细胞的经验表明，理想的免疫分离单克隆抗体应该：1）仅与一种类型的肾细胞相互作用，2）针对相对丰度存在的决定簇，3）无细胞毒性，4）是免疫球蛋白G。本文综述了可用于开发产生这种理想单克隆抗体的B淋巴细胞-骨髓瘤杂交细胞和使用细胞特异性单克隆抗体分离肾细胞的方法的一些概念和实践方面。
Several laboratories have described antibodies that are directed against cell surface antigens unique to different renal cell types. It can reasonably be assumed that each different type of renal cell possesses one or more unique antigenic determinants. In principle, it should be possible to prepare monoclonal antibodies against these cell-specific antigens and to use the antibodies as immunoaffinity reagents to isolate populations of specific renal cell types. We employed this approach in one instance to isolate canine cortical collecting tubule (CCT) cells. Approximately 10(7) canine CCT cells can be obtained from 5 g of canine renal cortex. This compares with an estimate of 10(3) to 10(4) CCT cells that can be reasonably obtained by microdissection. The availability of relatively large numbers of cells makes it possible to study in greater detail the cellular physiology and biochemistry of tubule-specific processes such as solute and water transport and hormone action. Our experiences in attempting to isolate canine CCT cells and other renal epithelia have indicated that the ideal monoclonal antibody for immunodissection should 1) interact with only one type of renal cell, 2) be directed against a determinant present in relative abundance, 3) be noncytotoxic, and 4) be an immunoglobulin G. Described in this review are some conceptual and practical aspects of the methods that can be used to develop B lymphocyte-myeloma hybrids producing such ideal monoclonal antibodies and to isolate renal cells using cell-specific monoclonal antibodies.