Sugimoto T, Sawada T, Negoro S, Kidowaki T, Morioka H, Matsumura T, Kemshead J T, Seeger R C
Cancer Res. 1985 Jan;45(1):358-64.
The expression of common acute lymphoblastic leukemia antigen (CALLA) on a human neuroblastoma cell line, SJ-N-CG, was demonstrated by indirect membrane immunofluorescence, complement-dependent cytotoxicity, and quantitative absorption, using two monoclonal antibodies (J-5 and BA-3) directed against CALLA. Immunoprecipitation of solubilized 125I-labeled membrane proteins from SJ-N-CG cells with J-5 antibody revealed a protein with a molecular weight of 100,000 as determined on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Morphological differentiation of SJ-N-CG cells could be induced in the presence of 2.0 mM dibutyryl adenosine 3'-5'-cyclic monophosphoric acid for 10 days of culture. Changes in cell surface membrane antigens associated with morphological differentiation were studied by indirect immunofluorescence and complement-dependent cytotoxicity using a panel of seven monoclonal antibodies. Increases in the antigens recognized by BA-2 (detecting leukemia-associated antigen), anti-Thy-1, and antibody 390 (Thy-1 antigen) were found in "differentiated cells," while those detected by BA-1 (B-cell-associated antigen) and J-5 (CALLA) were unchanged. In contrast, the antitransferrin receptor defined by B3/25 was inhibited, and expression of B7/21-defined la-like antigen was not induced. Kinetic studies on antigenic alterations showed that the expression of BA-2-defined antigen rose on Day 2 and remained at the same level until Day 10. The expression of CALLA was not changed from Days 2 to 10. The augmentation of Thy-1 antigen was noted on Day 4 and reached the maximum on Day 10. These results show that dibutyryl adenosine 3'-5'-cyclic monophosphoric acid is capable of inducing phenotypic changes in SJ-N-CG cells. The changes of expression of some antigens on exposure of cells to dibutyryl adenosine 3'-5'-cyclic monophosphoric acid may enable us to have a greater understanding of the differentiation of neuroblastoma to a more mature ganglioneuroblastoma phenotype.
利用两种针对普通急性淋巴细胞白血病抗原(CALLA)的单克隆抗体(J-5和BA-3),通过间接膜免疫荧光法、补体依赖细胞毒性法和定量吸收法,证实了人神经母细胞瘤细胞系SJ-N-CG上CALLA的表达。用J-5抗体对SJ-N-CG细胞中溶解的125I标记膜蛋白进行免疫沉淀,在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上测定显示分子量为100,000的一种蛋白。在2.0 mM二丁酰腺苷3'-5'-环磷酸存在下培养10天可诱导SJ-N-CG细胞发生形态分化。使用一组七种单克隆抗体,通过间接免疫荧光法和补体依赖细胞毒性法研究了与形态分化相关的细胞表面膜抗原的变化。在“分化细胞”中发现BA-2(检测白血病相关抗原)、抗Thy-1和抗体390(Thy-1抗原)识别的抗原增加,而BA-1(B细胞相关抗原)和J-5(CALLA)检测到的抗原没有变化。相反,由B3/25定义的抗转铁蛋白受体受到抑制,且未诱导出由B7/21定义的la样抗原的表达。对抗抗原改变的动力学研究表明,BA-2定义的抗原表达在第2天上升并在第10天保持在同一水平。CALLA的表达在第2天至第10天没有变化。Thy-1抗原的增加在第4天被注意到并在第10天达到最大值。这些结果表明二丁酰腺苷3'-5'-环磷酸能够诱导SJ-N-CG细胞发生表型变化。细胞暴露于二丁酰腺苷3'-5'-环磷酸时某些抗原表达的变化可能使我们对神经母细胞瘤向更成熟的神经节神经母细胞瘤表型的分化有更深入的了解。