Ramakrishnan S, Uckun F M, Houston L L
J Immunol. 1985 Nov;135(5):3616-22.
The ex vivo anti-leukemic efficacy and stem cell toxicity of two different T cell directed immunotoxins containing pokeweed antiviral protein (PAP) were studied by clonal assays. 5E9-11-PAP, an immunotoxin directed against human transferrin receptors, elicited a maximum leukemic cell kill of 3.9 logs. However, it was also toxic against normal pluripotent stem cells, and therefore is not a clinically useful purgative reagent. PAP conjugated to 3-A1, a monoclonal antibody directed against CD7 (T, p41), was more effective against leukemic T cells than 5E9-11-PAP and eliminated a maximum of 4.8 log of cells. 3A1-PAP was only slightly toxic to pluripotent stem cells: 13% of CFU-GEMM were lost after treatment with 3000 ng of 3A1-PAP/ml, a concentration that eliminated 99.96% of contaminating leukemic T cells from a 200-fold excess of normal bone marrow. Cryopreservation of treated cells by conventional methods did not affect the extreme selectivity and potency of 3A1-PAP. Incubation of 3A1-PAP with peripheral blood mononuclear cells resulted in the complete inhibition of phytohemagglutinin-induced mitogenic response, illustrating the possibility of using this immunotoxin as a potent anti-T cell reagent for prophylaxis against graft vs host disease in allogeneic BMT as well.
通过克隆分析研究了两种不同的含商陆抗病毒蛋白(PAP)的T细胞导向免疫毒素的体外抗白血病疗效和干细胞毒性。5E9-11-PAP是一种针对人转铁蛋白受体的免疫毒素,其最大白血病细胞杀伤率为3.9对数级。然而,它对正常多能干细胞也有毒性,因此不是一种临床上有用的净化试剂。与3-A1(一种针对CD7(T,p41)的单克隆抗体)偶联的PAP对白血病T细胞比5E9-11-PAP更有效,最多可清除4.8对数级的细胞。3A1-PAP对多能干细胞只有轻微毒性:用3000 ng/ml的3A1-PAP处理后,13%的粒-红-巨核-巨噬细胞集落形成单位(CFU-GEMM)丧失,该浓度可从200倍过量的正常骨髓中清除99.96%的污染白血病T细胞。用常规方法对处理后的细胞进行冷冻保存不会影响3A1-PAP的极端选择性和效力。3A1-PAP与外周血单个核细胞孵育导致植物血凝素诱导的有丝分裂反应完全受到抑制,这说明也有可能将这种免疫毒素用作预防异基因骨髓移植中移植物抗宿主病的强效抗T细胞试剂。