急性髓系白血病患者树突状细胞诱导抗自身白血病T细胞免疫的实验研究  

T-cell immunity against autologous leukemic cell mediated by in vitro bone marrow-derived dendritic cell from patients with acute myeloid leukemia in complete remission

作　　者：何学鹏[1] 尤胜国[1] 卞寿庚 葛薇[1] 李牧[1] 马双[1] 赵春华[1] 

机构地区：[1]中国医学科学院,中国协和医科大学血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中华血液学杂志》2001年第12期629-632,共4页Chinese Journal of Hematology

基　　金：卫生部重点项目科研基金资助项目 (WK3 2 0 0 0 1 3 4 )

摘　　要：目的　探讨自体白血病细胞裂解物 (ACL)冲击的完全缓解期的急性髓系白血病 (AML CR)患者骨髓细胞衍生的树突状细胞 (DC)体外能否刺激自体T细胞产生特异性抗AML细胞的细胞毒活性。方法　应用羊红细胞玫瑰花结程序从AML CR患者的骨髓中分离出去T细胞的骨髓单个核细胞 (TD BMNC) ,并培养在含联合细胞因子 (GM CSF、IL 4、SCF或TNF α)的条件下以产生DC ,并在培养的第 5天用ACL进行冲击。培养 7d后收获细胞 ,用流式细胞仪测定其成熟DC表型。同时 ,这些细胞与经抗CD3抗体激活过的自体T细胞在低浓度IL 2条件下共培养 7d ,以产生细胞毒性T淋巴细胞(CTL)。用乳酸脱氢酶释放实验测定溶细胞活性。结果　 1 2例AML CR患者培养的骨髓单个核细胞均分化为成熟DC。其中 6例完成CTL活性实验。当效∶靶 =2 0∶1时 ,ACL冲击的DC致敏的自体T细胞与单纯IL 2或IL 2加无抗原冲击的DC组比较对自体AML细胞有明显的杀伤活性 ,而对K562细胞均无明显影响 (P <0 .0 1 )。结论　用AML CR患者白细胞裂解物冲击的骨髓细胞衍生的DC体外致敏自体T细胞可以产生AML细胞相关抗原特异性的CTL。Objective To explore if a specific cytolytic T lymphocyte (CTL) response could be in vitro generated by using autogenous leukemic cells lysates (ACL) pulsed dendritic cells ( DCs ) from AML CR patients. Methods T cell depleted bone marrow mononuclear cells (TD BMNC) were isolated from AML CR patients with E rosetting procedure, and then were cultured in vitro in the presence of combined cytokines ( GM CSF, IL 4, SCF/TNF α) for generation of mature DCs, and pulsed with ACL on day 5. After 7 day culture, the cells were harvested and the expression of mature DC marker was determined by FACS. For generation of a specific CTL to the respective AML cells, the cells were co cultured with pre activated auto T lymphocytes by McAb anti CD3 in the presence of low concentration of IL 2 for 7 days. Cytotoxicity was determined with LDH release. Results Cultured TD BMNCs from 12 AML CR patients developed morphologic and phenotypic characteristics of mature DCs. CTL assay was performed in 6 out of the 12 samples. At effector/target ratio of 20:1, auto T lymphocytes primed with ACL pulsed DC exhibited significant killing activity to auto AML cells but not to K562 cells as compared with that stimulated by IL 2 alone or primed by non pulsed DC (P=0.001 ). Conclusion AML cell associated antigen specific CTL responses can be in vitro generated by priming auto T lymphocytes with ACL pulsed DCs. These findings might prove useful for immunotherapy of AML.

关 键 词：急性髓系白血病 树突状细胞 抗自身白血病 T细胞免疫 实验研究 

分 类 号：R733.7[医药卫生—肿瘤]