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鸡胚神经细胞间的黏附。I. 细胞间结合相关分子的免疫测定。

Adhesion among neural cells of the chick embryo. I. An immunological assay for molecules involved in cell-cell binding.

作者信息

Brackenbury R, Thiery J P, Rutishauser U, Edelman G M

出版信息

J Biol Chem. 1977 Oct 10;252(19):6835-40.

PMID:893445
Abstract

An immunologically based method for the quantitative assay of molecules involved in cell adhesion is described. Three observations served as a basis for this assay: (a) cells obtained by trypsinization of retinal tissue aggregated rapidly, provided they had been allowed to recover in culture from the dissociation process; (b) treatment of chick retinal cells with Fab' fragments from rabbit antibodies against these cells prevented their aggregation; and (c) incubation of these antibody fragments with antigens released by retinal cells in culture neutralized their ability to inhibit aggregation. The amount of neutralizing antigen was determined by measuring the rates of cell aggregation in the presence and absence of antibody and antigen using a particle counter. Although adhesion was inhibited by anti-retinal cell antibodies, it was not affected by lectins or anti-carbohydrate antibodies that also were bound to the cell surface. Together, the results suggest that the inhibition involved blockade or inactivation of particular cell surface molecules and that the retinal cell antigens capable of neutralizing the antibodies represented these molecules or their fragments. In the accompanying paper, we describe the use of this assay for the purification from culture supernatants of a cell surface molecule involved in cell to cell adhesion.

摘要

本文描述了一种基于免疫学的用于定量检测参与细胞黏附分子的方法。该检测基于以下三个观察结果:(a)通过胰蛋白酶消化视网膜组织获得的细胞,如果它们已在培养中从解离过程中恢复,则会迅速聚集;(b)用针对这些细胞的兔抗体的Fab'片段处理鸡视网膜细胞可防止其聚集;(c)将这些抗体片段与培养的视网膜细胞释放的抗原一起孵育可中和其抑制聚集的能力。通过使用粒子计数器测量在有和没有抗体及抗原存在的情况下细胞聚集的速率来确定中和抗原的量。虽然抗视网膜细胞抗体抑制了黏附,但它不受同样结合到细胞表面的凝集素或抗碳水化合物抗体的影响。总之,结果表明这种抑制涉及特定细胞表面分子的阻断或失活,并且能够中和抗体的视网膜细胞抗原代表了这些分子或其片段。在随附的论文中,我们描述了使用该检测方法从培养上清液中纯化参与细胞间黏附的细胞表面分子。

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