人急性髓系白血病HL-60来源的树突样细胞诱导的CTL体内外作用研究

The in vivo and in vitro response of cytotoxic T lymphocyte induced by HL-60-derived dendritic-like cell (HL-60DC)

目的 观察人急性髓系白血病HL 6 0来源的树突样细胞 (HL 6 0DC)对细胞毒T淋巴细胞 (CTL)的免疫调节作用。方法 采用PKC激活剂佛波酯 (PMA)及细胞因子GM CSF、IL 4、TNF α在体外诱导培养HL 6 0细胞 ,获得具有树突状细胞形态、表型 (CD1a、CD80、CD86、RelB阳性表达 )的HL 6 0DC ,观察HL 6 0DC诱导的CTL体外抗瘤效应 ,并建立人白血病HL 6 0 SCID(严重联合免疫缺陷 )小鼠异种移植模型进行过继治疗。结果 联合GM CSF、TNF α及PMA处理 7～ 11d ,获得的HL 6 0DC体外激活的CTL在体外对HL 6 0细胞有显著的细胞毒活性。CTL以 5∶1效靶比多次腹腔回输 ,能够明显延长荷瘤鼠存活时间 ,降低瘤块重量 ,减轻肝、脾、骨髓受肿瘤浸润程度 ,但流式细胞检测发现部分受治疗的存活小鼠有微小病灶的存在。结论 人急性髓系白血病HL 6 0来源的树突样细胞诱导的CTL在体内外有一定的抗瘤效应。

Objective To demonstrate the immune effects of HL-60-derived dendritic-like cell (HL-60DC) on cytotoxic T lymphocyte (CTL). Methods After being treated by PMA, GM-CSF, TNF-α and IL-4, HL-60 cells were differentiated into HL-60DC with the phenotypic DC characteristics including morphologic feature, surface antigens CD1a +/CD86 +/CD80 + and RelB expression as well. The anti-tumor effects of CTL induced by HL-60DC were tested in vitro and in the adoptive therapy experiments of HL-60/SCID mice. Results The CTLs induced by HL-60DC which was treated by PMA and GM-CSF plus TNF-α for 7-11 days lysed HL-60 cell in vitro. Multiple intraperitoneal transfusions of CTL (effector∶target=5∶1) into HL-60/SCID mice improved survival of the treated animals and reduced the infiltrations of leukemic cells in liver, spleen, bone marrow and the weight of tumor. Flow cytometric analysis showed a minimal residual disease (MRD) was present in some of the treated mice. Conclusion It was confirmed that the anti-tumor effects of CTL induced by HL-60DC was present both in vitro and in HL-60/SCID mice xenograft model.