Kreitman R J, Chaudhary V K, Waldmann T, Willingham M C, FitzGerald D J, Pastan I
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1990 Nov;87(21):8291-5. doi: 10.1073/pnas.87.21.8291.
Anti-Tac(Fv)-PE40 is a recombinant single-chain immunotoxin containing the heavy and light variable regions of the anti-Tac monoclonal antibody fused to a mutant form of Pseudomonas exotoxin (PE). Anti-Tac binds to the p55 subunit of the human interleukin 2 (IL-2) receptor, and anti-Tac(Fv)-PE40 kills human or monkey cell lines that contain either the intact IL-2 receptor or its p55 subunit alone. To assess the usefulness of anti-Tac(Fv)-PE40 in treatment of IL-2 receptor-positive leukemia, we tested peripheral blood mononuclear cells from six patients with adult T-cell leukemia. In each of the six patients, anti-Tac(Fv)-PE40 was extremely cytotoxic to the malignant cells. Metabolic activity and sensitivity of the fresh cells improved when a small amount of IL-2 (10 units per ml) was present during incubation. The toxin concentration necessary to inhibit protein synthesis by 50% after 16-hr incubation of cells with immunotoxin varied from 1.6 to 16 ng/ml (2.5-25 x 10(-11) M). In every case, binding was by means of the Tac antigen because anti-Tac(Fv)-PE40 cytotoxicity was prevented by adding excess anti-Tac antibody. Moreover, anti-Tac alone or an inactive mutant of anti-Tac(Fv)-PE40 without ADP-ribosylation activity had very little cytotoxic activity. Peripheral blood mononuclear cells from normal controls, from a patient with Tac-negative leukemia, and from adult T-cell leukemia patients without significant peripheral blood involvement were not sensitive to anti-Tac(Fv)-PE40. These results indicate that anti-Tac(Fv)-PE40 is a potent cytotoxin against adult T-cell leukemia cells in vitro and warrants clinical testing.
抗 Tac(Fv)-PE40 是一种重组单链免疫毒素,它包含抗 Tac 单克隆抗体的重链和轻链可变区,并与铜绿假单胞菌外毒素(PE)的突变形式融合。抗 Tac 可与人白细胞介素 2(IL-2)受体的 p55 亚基结合,抗 Tac(Fv)-PE40 可杀死含有完整 IL-2 受体或仅其 p55 亚基的人或猴细胞系。为了评估抗 Tac(Fv)-PE40 在治疗 IL-2 受体阳性白血病中的效用,我们检测了 6 例成人 T 细胞白血病患者的外周血单个核细胞。在这 6 例患者中,抗 Tac(Fv)-PE40 对恶性细胞具有极强的细胞毒性。当在孵育过程中存在少量 IL-2(每毫升 10 单位)时,新鲜细胞的代谢活性和敏感性有所提高。细胞与免疫毒素孵育 16 小时后,抑制蛋白质合成 50%所需的毒素浓度在 1.6 至 16 纳克/毫升(2.5 - 25×10⁻¹¹ 摩尔/升)之间。在每种情况下,结合都是通过 Tac 抗原进行的,因为加入过量的抗 Tac 抗体可阻止抗 Tac(Fv)-PE40 的细胞毒性。此外,单独的抗 Tac 或没有 ADP - 核糖基化活性的抗 Tac(Fv)-PE40 无活性突变体几乎没有细胞毒性活性。来自正常对照、Tac 阴性白血病患者以及无明显外周血受累的成人 T 细胞白血病患者的外周血单个核细胞对抗 Tac(Fv)-PE40 不敏感。这些结果表明,抗 Tac(Fv)-PE40 在体外是一种针对成人 T 细胞白血病细胞的强效细胞毒素,值得进行临床试验。