Bhattacharya-Chatterjee M, Pride M W, Seon B K, Kohler H
J Immunol. 1987 Aug 15;139(4):1354-60.
A murine monoclonal anti-tumor antibody termed SN2 (Ab1), isotype IgG1-kappa, that defines a unique human T cell leukemia-associated cell-surface glycoprotein, gp37 (m.w. 37,000), was used to generate monoclonal anti-idiotype antibodies (Ab2) in syngeneic BALB/c mice. The Ab2 were screened on the basis of their binding to the F(ab')2 fragments of SN2 and not to the F(ab')2 of pooled normal BALB/c mice sera IgG1 or to an unrelated BALB/c monoclonal antibody of the same isotype. Fifteen Ab2, obtained from two fusions, were specific for the SN2 idiotope and not against isotype or allotype determinants. To find out whether these Ab2 are directed against the paratope of SN2, the binding of radiolabeled SN2 to leukemic MOLT-4 and JM cells which contain gp37 as a surface constituent was studied in the presence of these anti-idiotopes. Clone 4EA2 inhibited the binding 100% at a concentration of 50 ng and 4DC6 inhibited 90% at a concentration of 250 ng. A third clone 4DD6 gave about 50% inhibition. Similar was the inhibition of SN2 binding to insolubilized MOLT-4 antigen or cell membrane preparation. The binding of SN2 (Ab1) to 4EA2 and 4DC6 was also inhibited by semipurified preparation of gp37 antigen. These results demonstrate that at least two of the anti-idiotope antibodies are binding either at or near the binding site idiotope of SN2. Next, the purified Ab2 was used to immunize syngeneic mice to induce antibody binding to MOLT-4 cells or gp37. Sera from mice immunized with 4EA2 and 4DC6 coupled to keyhole limpet hemocyanin contained antibodies which bind to semipurified gp37 antigen and MOLT-4 cells. Immune sera inhibited the binding of iodinated Ab2 and Ab1 indicating that an anti-anti-idiotopic antibody (Ab3) in mice shares idiotopes with Ab1 (SN2). Also, the binding of iodinated Ab2 to Ab1 was inhibited by rabbit antisera specific for gp37. Collectively, these data suggest that anti-idiotype antibodies 4EA2 and 4DC6 may be useful in the generation of idiotype vaccines against human T cell leukemia.
一种名为SN2（Ab1）的鼠单克隆抗肿瘤抗体，同种型为IgG1-κ，它定义了一种独特的与人类T细胞白血病相关的细胞表面糖蛋白gp37（分子量37,000），被用于在同基因BALB/c小鼠中产生单克隆抗独特型抗体（Ab2）。Ab2根据它们与SN2的F(ab')2片段结合而不与正常BALB/c小鼠血清IgG1的F(ab')2或同一种同种型的无关BALB/c单克隆抗体结合的特性进行筛选。从两次融合中获得的15种Ab2对SN2独特型具有特异性，而不针对同种型或异型决定簇。为了确定这些Ab2是否针对SN2的互补位，在这些抗独特型抗体存在的情况下，研究了放射性标记的SN2与含有gp37作为表面成分的白血病MOLT-4和JM细胞的结合。克隆4EA2在浓度为50 ng时100%抑制结合，4DC6在浓度为250 ng时90%抑制结合。第三个克隆4DD6产生约50%的抑制。SN2与不溶性MOLT-4抗原或细胞膜制剂的结合抑制情况类似。gp37抗原的半纯化制剂也抑制了SN(Ab1)与4EA2和4DC6的结合。这些结果表明，至少两种抗独特型抗体在SN2独特型结合位点或其附近结合。接下来，将纯化的Ab2用于免疫同基因小鼠，以诱导抗体与MOLT-4细胞或gp37结合。用与钥孔血蓝蛋白偶联的4EA2和4DC6免疫的小鼠血清中含有与半纯化gp37抗原和MOLT-4细胞结合的抗体。免疫血清抑制了碘化Ab2和Ab1的结合，表明小鼠中的抗抗独特型抗体（Ab3）与Ab1（SN2）共享独特型。此外，gp37特异性兔抗血清抑制了碘化Ab2与Ab1的结合。总体而言，这些数据表明抗独特型抗体4EA2和4DC6可能有助于生成针对人类T细胞白血病的独特型疫苗。
