Cell surface reactive human monoclonal antibody directed to human melanoma-associated gangliosides.
Cell surface reactive human monoclonal antibody directed to human melanoma-associated gangliosides.
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针对人黑色素瘤相关神经节苷脂的细胞表面反应性人单克隆抗体。
DOI：
10.1097/00008390-199311000-00004
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发表时间：
1993
影响因子：
2.2
通讯作者：
Seigler,HF
中科院分区：
文献类型：
--
作者：
Abdel-Wahab,Z;Li,WP;Darrow,T;Nudelman,ED;Towell,A;Seigler,HF
从恶性黑色素瘤手术标本中分离出的淋巴结细胞与异种骨髓瘤细胞系SHMD-33融合，制备了一种IgM人源单抗。该抗体命名为7c11。活细胞免疫荧光和吸收实验表明，E8能与人黑色素瘤细胞表面抗原发生反应。单抗7c11。E8与DSI、SPG、GM4、GM3和GD3反应，而与GD2、GM1、GM2、GD1a、GD1b、GT1b和一些中性糖鞘蛋白不反应。因此，单抗的主要结合表位是末端的N-乙酰神经氨酸2-3Gal，通过与神经纤维的[β]1-1键或与葡萄糖或氨基葡萄糖的[β]-4键相连。免疫组织化学分析显示，7c11。E8抗原表达于所有黑色素瘤组织，以及少数结肠癌、正常结肠、皮肤、脊髓、肾脏和肝脏组织。然而，其他正常器官如乳腺、肺、小肠、胃和淋巴结不与单抗反应。在人血清存在的情况下，该抗体在补体依赖的细胞毒性(CDCC)试验中启动了对黑色素瘤细胞的强烈裂解。这项研究表明，在筛选方案中，使用活细胞分析方法可以分离针对细胞表面抗原的人源单抗。7c11的优先结合。E8对黑色素瘤组织和与两种主要黑色素瘤神经节苷脂(GM3和GD3)的反应性表明7c11。E8可能为恶性黑色素瘤的诊断和治疗提供有用的试剂。
An IgM human monoclonal antibody (human MAb) was generated by fusing lymph node cells Isolated from a surgical specimen of malignant melanoma with the heteromyeloma cell line SHMD-33. The antibody, designated 7c11. e8, reacted with surface antigens on human melanoma cells as shown by live cell immunofluorescence and absorption assays. The MAb 7c11. e8 reacted with DSI, SPG, GM4, GM3 and GD3 In enzyme-linked Immunosorbent assays (ELISA), and did not react with GD2, GM1, GM2, GD1a, GD1b, GT1b and a number of neutral glycosphingollplds. The main binding epitope for the MAb was, therefore, the terminal N-acetylneuramlnlc acid 2-3 Gal linked by a [beta] 1-1 bond to the ceramlde, or a [beta]-4 bond to glucose or glucosamine. As shown by immunohlstochemlcal assays, 7c11. e8 antigen was expressed on all melanoma tumour tissues, and on a few samples of colon carcinoma, normal colon, skin, spinal cord, kidney and liver. However, other normal organs such as breast, lung, small Intestine, stomach and lymph nodes did not react with the MAb. In the presence of human serum the antibody Initiated a strong lysis of melanoma tumour cells In complement-dependent cellular cytotoxicity (CDCC) assays. This study demonstrates that It Is possible to Isolate human monoclonal antibodies directed to cell surface antigens using viable cell assays In the screening protocol. The preferential binding of 7c11. e8 to melanoma tissues and the reactivity with two of the major melanoma gangliosides (GM3 and GD3) suggest that 7c11. e8 may provide a useful reagent for diagnosis and therapy of malignant melanoma.