Natoli E J, Livingston P O, Pukel C S, Lloyd K O, Wiegandt H, Szalay J, Oettgen H F, Old L J
Cancer Res. 1986 Aug;46(8):4116-20.
Antisera reactive with the ganglioside GM2 were raised by immunizing C57BL/6 mice with the C57BL/6 melanoma JB-RH. Fusion with NS-1 was performed using splenic mononuclear cells from a mouse with high antibody titer. An immunoglobulin M monoclonal antibody (monoclonal antibody 5-3) was identified which was reactive with an antigen that was resistant to heat, trypsin, and Pronase. A panel of purified glycolipids was used to determine the specificity of monoclonal antibody 5-3. Reactivity was restricted to N-acetyl- and N-glycolyl-GM2. No reactivity was detected with asialo-GM2 or other gangliosides. Monoclonal antibody 5-3 was used to define the expression of GM2 on the cell surface of cultured human normal and malignant cells. Reactivity was seen with cell lines derived from 8 of 8 astrocytomas, 5 of 5 neuroblastomas, 7 of 9 sarcomas, 4 of 18 human melanomas, 2 of 4 murine melanomas, 4 of 37 epithelial cancers and with 0 of 6 skin fibroblast and 0 of 2 brain fibroblast lines. GM2, like GD2 and GD3, appears to be a differentiation antigen largely restricted to cells of neuroectodermal origin.
通过用C57BL/6黑色素瘤JB-RH免疫C57BL/6小鼠，制备了与神经节苷脂GM2反应的抗血清。使用来自抗体滴度高的小鼠的脾单核细胞与NS-1进行融合。鉴定出一种免疫球蛋白M单克隆抗体（单克隆抗体5-3），它与一种对热、胰蛋白酶和链霉蛋白酶有抗性的抗原反应。使用一组纯化的糖脂来确定单克隆抗体5-3的特异性。反应性仅限于N-乙酰基和N-糖基化GM2。未检测到与脱唾液酸GM2或其他神经节苷脂的反应性。单克隆抗体5-3用于确定GM2在培养的人正常细胞和恶性细胞表面的表达。在来自8例星形细胞瘤中的8例、5例神经母细胞瘤中的5例、9例肉瘤中的7例、18例人黑色素瘤中的4例、4例鼠黑色素瘤中的2例、37例上皮癌中的4例的细胞系中观察到反应性，而在6例皮肤成纤维细胞系中的0例和2例脑成纤维细胞系中的0例中未观察到反应性。GM2与GD2和GD3一样，似乎是一种主要限于神经外胚层来源细胞的分化抗原。
