CD3AK细胞对耐药白血病细胞的体外净化作用

目的 :观察CD3 AK细胞对耐药的白血病细胞系及慢性髓细胞白血病 (chronicmyelogenousleukemia ,CML)急变患者原代肿瘤细胞的体外净化作用。方法 :采用固化的抗CD3单克隆抗体联合小剂量IL 2诱导CD3 AK细胞 ;MTT法观察CD3 AK细胞对K5 6 2、HL6 0及其耐药株的细胞毒活性 ;肿瘤细胞集落培养 (tumorcolonyassay ,TCA)观察CD3 AK细胞对K5 6 2、HL6 0及其耐药株集落形成的抑制作用 ;流式细胞仪 (flowcytometry ,FCM)检测耐药的CML急变患者原代细胞经CD3 AK细胞净化后Pgp阳性细胞的比例变化。结果 :MT法显示CD3 AK细胞在体外对K5 6 2细胞、HL6 0细胞及其耐药株有相似的杀伤作用 ;集落培养观察CD3 AK细胞对HL6 0细胞株及其耐药株的集落形成均有较强的抑制作用 ;FCM结果显示耐药CML原代细胞经CD3 AK细胞净化后Pgp阳性细胞比例下降 2 3 2 0 %。结论 :CD3 AK细胞在体外对耐药白血病细胞株及耐药白血病原代细胞均有较强的净化作用。

Using of anti-CD3 monoclonal antibody activated killer cells(CD3AK) in purging of multi-drug resistant leukemic cell lines and leukemic cells from chronic myelogenous leukemia patients.

Objective:To investigate the probability of using anti CD3 monoclonal antibody activated killer cells (CD 3AK) as a method to purge multi drug resistant leukemic cell lines and leukemic cells from chronic myelogenous leukemia patients. Methods:The immobilized anti CD3 monoclonal antibody and low dosed IL 2 were used to active CD 3AK. The ability of CD 3AK to purge K562, HL60 and their multi drug resistant subclones in vitro was evaluated by MTT (Thiazolyl blue) method. The percentages of Pgp positive cells was analysed by FCM. Tumor Colony Assay (TCA) was used to assay the influence of CD 3AK on colony forming capability of HL60 cell line and its multi drug resistant subclone, HL60/ADM.Results:CD 3AK possessed similar cytotoxic activity to K562 cell line, HL60 cell line and their multi drug resistant subclones as determined by MTT. After leukemic cells from chronic myelogenous leukemiablast crisis (CML BC) patients were cocultured with allogeneic CD 3AK for six hours, the percentages of Pgp positive cells decreased 23 20% assayed by FCM. TCA also demonstrated that CD 3AK has similar capability to inhibite colony forming of HL60 cell line and HL60/ADM. Conclusion:CD 3AK has significant cytotoxic activity on multi drug resistant leukemic cell lines and leukemic cells from CML BC patients.