Harada R, Takahashi N, Owaki I, Kannagi R, Endo N, Morita N, Inoue M
Department of Clinical Science and Laboratory Medicine, School of Medicine, Kyoto University, Japan.
Igaku Kenkyu. 1992 Feb;62(1):1-18.
A human monoclonal antibody, ll-50 (IgM, lambda), was generated, which reacted specifically with a major of glycolipid present in LS174T colon cancer cells. The glycolipid antigen which reacted with the ll-50 antibody was expected to four sugar residues from its TLC mobility, and it was ascertained that the glycolipid antigen which reacted with ll-50 antibody might be Lc4 antigen [Gal beta 1----3 GLcNAc beta 1----3 Gal beta 1----4 Glc beta 1----1 Cer] judging from TLC immunostaining and ELISA when the reactivity of ll-50 antibody was tested using various pure glycolipids in 3-5 sugar residues as an antigen. Sera in patients with malignant disorders and healthy individuals were analyzed by Sandwich assay of immobilized and biotinylated ll-50 antibody. The serum of the Lc4 antigen recognized by ll-50 antibody was significantly higher in patients with malignant disorders than that in healthy individuals (p less than 0.05). Three mouse monoclonal anti-idiotype antibodies, G3, B3 and C5 (all IgG1), were generated by the immunization of BALB/c mice with ll-50 antibody. These anti-idiotype antibodies specifically bound to to human monoclonal antibody, ll-50 and had a significant inhibitory activity towards the binding of ll-50 antibody to the Lc4 antigen. This indicated that these anti-idiotype antibodies, G3, B3, and C5, were paratope-related anti-idiotype antibodies. G3, B3, and C5 were expected to define the nearest idiotope because they could mutually inhibit ll-50 antibody. Sera in patients with malignant disorders and healthy individuals were analyzed by Sandwich assay of immobilized and biotinylated anti-idiotype antibodies, G3, B3, and C5. As to the ll-50 like antibodies defined by C5 (Id-C5+), the mean serum level in patients with malignant disorders was significantly higher than that in healthy individuals (p less than 0.05). As to the ll-50 like antibodies defined by B3 (Id-B3+), the mean serum level in patients with malignant disorders was significantly higher than that in healthy individuals.(ABSTRACT TRUNCATED AT 400 WORDS)
产生了一种人源单克隆抗体ll-50（IgM，λ），它能与LS174T结肠癌细胞中存在的一种主要糖脂特异性反应。根据薄层层析迁移率，与ll-50抗体反应的糖脂抗原预计有四个糖残基。当使用含有3至5个糖残基的各种纯糖脂作为抗原，通过薄层层析免疫染色和酶联免疫吸附测定来检测ll-50抗体的反应性时，确定与ll-50抗体反应的糖脂抗原可能是Lc4抗原[半乳糖β1----3 N-乙酰葡糖胺β1----3半乳糖β1----4葡萄糖β1----1神经酰胺]。使用固定化和生物素化的ll-50抗体的夹心测定法分析了恶性疾病患者和健康个体的血清。ll-50抗体识别的Lc4抗原的血清水平在恶性疾病患者中显著高于健康个体（p<0.05）。通过用ll-50抗体免疫BALB/c小鼠，产生了三种小鼠单克隆抗独特型抗体G3、B3和C5（均为IgG1）。这些抗独特型抗体特异性结合人源单克隆抗体ll-50，并对ll-50抗体与Lc4抗原的结合具有显著抑制活性。这表明这些抗独特型抗体G3、B3和C5是与互补位相关的抗独特型抗体。由于G3、B3和C5能相互抑制ll-50抗体，预计它们能确定最接近的独特型。使用固定化和生物素化的抗独特型抗体G3、B3和C5的夹心测定法分析了恶性疾病患者和健康个体的血清。对于由C5定义的ll-50样抗体（Id-C5+），恶性疾病患者的平均血清水平显著高于健康个体（p<0.05）。对于由B3定义的ll-50样抗体（Id-B3+），恶性疾病患者的平均血清水平显著高于健康个体。（摘要截短于400字）
