Bonardi M A, French R R, Amlot P, Gromo G, Modena D, Glennie M J
Lymphoma Research Unit, Tenovus Laboratory, General Hospital, Southampton, United Kingdom.
Cancer Res. 1993 Jul 1;53(13):3015-21.
A panel of bispecific F(ab')2 antibodies (BsAb) have been constructed for delivering the ribosome-inactivating protein saporin to human B cell lymphoma. Each derivative was prepared with specificity for saporin and CD19, CD22, CD37, or immunoglobulin. In vitro studies measuring inhibition of [3H]leucine uptake by cultured Daudi and Raji cells demonstrated that, despite all BsAb capturing saporin on the cell surface, BsAb targeting through CD22 were far more cytotoxic than those functioning via CD19, CD37, or surface immunoglobulin. This exceptional activity of the CD22-specific BsAb appears to derive from its ability to deliver and accumulate saporin inside the target cells. Further studies showed that four CD22-specific BsAb all performed with equal potency and were able to increase saporin toxicity (50% inhibitory concentration) up to 1000-fold, from 2 x 10(-7) M to 2 x 10(-10) M. Pairs of anti-CD22 BsAb which recognized different nonblocking epitopes on the saporin molecule were able to bind saporin more avidly to the target cell and, as a consequence, increased cytotoxicity by at least an additional 10-fold, resulting in 50% inhibitory concentration for protein synthesis of 2 x 10(-11) M. These results suggest that selected combinations of BsAb which bind cooperatively to a toxin and the cell surface may provide an efficient way of delivering toxins to unwanted cells in patients.
已构建了一组双特异性F(ab')2抗体(BsAb),用于将核糖体失活蛋白皂草素递送至人B细胞淋巴瘤。每种衍生物都针对皂草素和CD19、CD22、CD37或免疫球蛋白制备。通过测量培养的Daudi细胞和Raji细胞对[3H]亮氨酸摄取的抑制作用进行的体外研究表明,尽管所有BsAb都能在细胞表面捕获皂草素,但靶向CD22的BsAb比通过CD19、CD37或表面免疫球蛋白发挥作用的BsAb具有更强的细胞毒性。这种CD22特异性BsAb的特殊活性似乎源于其将皂草素递送至靶细胞内并在其中积累的能力。进一步的研究表明,四种CD22特异性BsAb的效力均相同,并且能够将皂草素毒性(50%抑制浓度)提高至1000倍,从2×10(-7)M降至2×10(-10)M。识别皂草素分子上不同非阻断表位的抗CD22 BsAb对能够更有效地将皂草素结合至靶细胞,结果使细胞毒性至少再增加10倍,导致蛋白质合成的50%抑制浓度达到2×10(-11)M。这些结果表明,与毒素和细胞表面协同结合的BsAb的特定组合可能为将毒素递送至患者体内不需要的细胞提供一种有效的方法。