公开(公告)号：US4507391A

公开(公告)日：1985-03-26

申请号：US365065

申请日：1982-04-02

申请人： Clifford S. Pukel ,  Kenneth O. Lloyd ,  Luiz R. Travassos ,  Wolfgang G. Dippold ,  Herbert F. Oettgen ,  Lloyd J. Old

发明人： Clifford S. Pukel ,  Kenneth O. Lloyd ,  Luiz R. Travassos ,  Wolfgang G. Dippold ,  Herbert F. Oettgen ,  Lloyd J. Old

IPC分类号： C12Q1/04 ,  G01N33/53 ,  G01N33/574 ,  G01N33/54 ,  C12N15/00

CPC分类号： G01N33/5743 ,  G01N33/5308 ,  Y10S436/804 ,  Y10S436/811 ,  Y10S436/813 ,  Y10S436/815 ,  Y10S436/828

摘要： Mouse monoclonal antibody AbR.sub.24 (Dippold et al., Proc. Natl. Acad. Sci. 77:6114-6118, 1980) has a high degree of specificity for human melanoma cells when tested on viable cultured cells using the PA-MHA serological assay. The antigen detected by this antibody has been isolated from melanoma cells and shown to be G.sub.D3 ganglioside by compositional and partial structural analysis and by comparison with authentic G.sub.D3 by thin layer chromatography (TLC). AbR.sub.24 reacts with authentic G.sub.D3, but not with any other ganglioside tested. Using TLC and reactivity with AbR.sub.24, a wide range of cells and tissues was examined for the presence of G.sub.D3. A new serological assay, termed glycolipid-mediated immune adherence (GMIA), was devised for assaying the reactivity of AbR.sub.24 with gangliosides. Melanomas (cultured cells or tumor tissue) were shown to have T.sub.D3 and G.sub.M3 as major gangliosides. Other cells and tissues examined also contained G.sub.D3, but usually only in low amounts. Melanomas (and MOLT-4, a T-cell line) were characterized by a simplified ganglioside profile with G.sub.D3 and G.sub.M3 as major components. The apparent discrepancy between the ubiquitous presence of G.sub.D3 and the serological specificity of AbR.sub.24 for melanoma cells can be explained in terms of localization and concentration of G.sub.D3 in different cells.

摘要翻译： 当使用PA-MHA血清学测定法在活细胞培养细胞上测试时，小鼠单克隆抗体AbR24（Dippold等人，Proc.Natl.Acad.Sci.77：6114-6118,1980）对人黑素瘤细胞具有高度的特异性。 由该抗体检测到的抗原已从黑素瘤细胞中分离出来，通过组成和部分结构分析显示为GD3神经节苷脂，并通过薄层色谱（TLC）与真实的GD3进行比较。 AbR24与正常的GD3反应，但不与任何其他神经节苷脂检测。 使用TLC和与AbR24的反应性，检查广泛的细胞和组织的GD3的存在。 设计了一种称为糖脂介导的免疫粘附（GMIA）的血清学测定法，用于测定AbR24与神经节苷脂的反应性。 黑色素瘤（培养细胞或肿瘤组织）显示具有TD3和GM3作为主要神经节苷脂。 检查的其他细胞和组织也含有GD3，但通常只有少量。 黑色素瘤（和MOLT-4，T细胞系）以GD3和GM3为主要成分的简化神经节苷脂特征表征。 GD3的普遍存在与黑素瘤细胞的AbR24的血清学特异性之间的明显差异可以根据不同细胞中GD3的定位和浓度来解释。