Monoclonal antibodies directed to chemically synthesized lactogangliotetraosylceramide, a leukemia-associated antigen having a novel branching structure.
Monoclonal antibodies directed to chemically synthesized lactogangliotetraosylceramide, a leukemia-associated antigen having a novel branching structure.
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针对化学合成的乳神经节四糖神经酰胺的单克隆抗体，乳神经节四糖神经酰胺是一种具有新型分支结构的白血病相关抗原。
DOI：
10.1016/s0021-9258(19)75794-9
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
R. Kannagi
中科院分区：
文献类型：
--
作者：
K. Shigeta;Y. Ito;T. Ogawa;Y. Kirihata;S. Hakomori;R. Kannagi
处于未分化状态的鼠白血病细胞（M1）的特征在于存在与癌症相关的乳神经节系列糖脂，其中一种糖脂被鉴定为乳神经节四糖基神经酰胺（LcGg 4），其通过GlcNAc β 1- 3和GalNAc β 1- 4键在乳糖基神经酰胺的II-Gal处具有新的分支，如下所示（Kannagi，R.，Levery，S.B.，和Hakomori，S.（1984）生物化学杂志，259，8444-8451）：GalNAc β 1-4Gal β 1-4Glc β 1-1Cer GlcNac β 1-3由于这种糖脂是非常少量的组分，因此难以获得足够的纯化糖脂用于制备单克隆抗体。我们开发了一种方法来化学合成这种糖脂，使用乳糖单位，神经酰胺单位，和两个己糖胺供体作为底物，并使合成的糖脂可作为免疫原。我们获得的两个单克隆抗体（YI 328 -18和YI 328 -51，均为IgG 3）特异性识别新的分支结构，并且与神经节三己糖神经酰胺或乳糖三己糖神经酰胺没有交叉反应性。因此，发现抗体是检测未分化的M1白血病细胞中表达的乳神经节四糖神经酰胺的有用探针，其在诱导分化时消失。本研究的结果表明，一种新的策略，建立单克隆抗体针对新的次要糖脂标记物或其人工设计的类似物，采用化学合成的糖脂抗原。
Murine leukemia cells (M1), in their undifferentiated state, have been characterized by the presence of cancer-associated lactoganglio-series glycolipids, one of which was identified as lactogangliotetraosylceramide (LcGg4) having a novel branching at the II-Gal of lactosylceramide through GlcNAc beta 1----3 and GalNAc beta 1----4 linkage, as shown below (Kannagi, R., Levery, S.B., and Hakomori, S. (1984) J. Biol. Chem., 259, 8444-8451): GalNAc beta 1----4 Gal beta 1----4Glc beta 1----1Cer GlcNac beta 1----3 Since this glycolipid is a very minor component, it has been difficult to obtain enough of the purified glycolipid for the preparation of a monoclonal antibody. We developed a method to chemically synthesize this glycolipid using a lactose unit, a ceramide unit, and two hexosamine donors as synthons and made the synthetic glycolipid available as an immunogen. The two monoclonal antibodies we obtained (YI328-18 and YI328-51, both IgG3) specifically recognized the novel branching structure and had no cross-reactivity with gangliotriaosylceramide or lactotriaosylceramide. Thus, the antibodies were found to be useful probes to detect lactogangliotetraosylceramide expressed in undifferentiated M1 leukemia cells, which disappears on induced differentiation. The results of this study indicate a new strategy to establish monoclonal antibody directed to novel minor glycolipid markers or their artificially designed analogs, employing chemically synthesized glycolipid antigens.