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通过针对盘基网柄菌凝集素I的特异性单价抗体确定膜凝集素在盘基网柄菌聚集中的作用。

The role of membrane lectins in Dictyostelium discoideum aggregation as ascertained by specific univalent antibodies against discoidin I.

作者信息

Cano A, Pestaña A

出版信息

J Cell Biochem. 1984;25(1):31-43. doi: 10.1002/jcb.240250104.

Abstract

Antibodies against pure discoidin I have been used as a tool to ascertain the role of this lectin in aggregation of Dictyostelium discoideum. Discoidin I is widely expressed over the cell surface of aggregation-competent AX-2 cells, as ascertained by indirect immunofluorescence with specific (antidiscoidin I) antibodies. Univalent antidiscoidin I antibodies (Fab fragments) inhibit the aggregation-specific intercellular adhesion of D discoideum AX-2 cells in an in vitro assay. This inhibition depends on antibody concentration and cell density; a 50% inhibition of cell aggregation was obtained at antidiscoidin I Fab concentration of 4.5 mg/ml and 1 X 10(6) cells/ml. Aggregation and morphogenesis on solid support is also effectively inhibited when AX-2 cells are starved in the presence of antidiscoidin I Fab fragments. The inhibition of morphogenesis is also dose dependent and more effective than in the in vitro assay. No inhibition of aggregation either in the in vitro assay or on morphogenesis on solid support was observed with preimmune Fab fragments at any of the concentrations tested (up to 9.6 mg/ml).

摘要

抗纯盘基网柄菌凝集素I的抗体已被用作一种工具,以确定这种凝集素在盘基网柄菌聚集过程中的作用。通过用特异性(抗盘基网柄菌凝集素I)抗体进行间接免疫荧光测定,发现盘基网柄菌凝集素I在具有聚集能力的AX-2细胞的细胞表面广泛表达。单价抗盘基网柄菌凝集素I抗体(Fab片段)在体外试验中可抑制盘基网柄菌AX-2细胞的聚集特异性细胞间黏附。这种抑制作用取决于抗体浓度和细胞密度;在抗盘基网柄菌凝集素I Fab浓度为4.5 mg/ml和细胞密度为1×10⁶个细胞/ml时,细胞聚集受到50%的抑制。当AX-2细胞在抗盘基网柄菌凝集素I Fab片段存在的情况下饥饿培养时,在固体支持物上的聚集和形态发生也受到有效抑制。形态发生的抑制也是剂量依赖性的,并且比体外试验更有效。在任何测试浓度(高达9.6 mg/ml)下,用免疫前Fab片段在体外试验或固体支持物上的形态发生试验中均未观察到聚集抑制。

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