多发性骨髓瘤患者树突状细胞介导的特异性抗瘤活性的体外诱导

Induction of specific cytotoxic T lymphocytes mediated by dendritic cells in multiple myeloma patients in vitro

目的 :观察U2 6 6 细胞及其可溶性抗原激发的多发性骨髓瘤 (MM)患者树突状细胞 (DC)体外诱导U2 6 6 特异性CTL的作用。方法 :将MM患者外周血来源的单核细胞在rhGM CSF 80 0U ml与IFNα 6 0 0U ml条件下利用无血清技术培养生成DC ,应用丝裂霉素C处理的U2 6 6 细胞及用U2 6 6 细胞制备的可溶性抗原预刺激DC ,然后与自体淋巴细胞共同孵育 5～ 7d以诱生特异性CTL ,采用MTT法检测对U2 6 6 细胞的特异性杀伤效果。结果 :MM患者外周血单核细胞在GM CSF IFNα条件下培养 8d后生成具有典型特征的DC ,高度表达CD86、CD5 4及MHCII类分子HLA DR。应用MTT法检测U2 6 6 细胞及其可溶性抗原激发的DC诱导特异性CTL对靶细胞U2 6 6 的杀伤率分别为 2 1 2 %± 5 4 %和 2 8 0 %± 7 6 % ,对照组未用抗原刺激组都为 11 7%±4 3%。而以抗原直接刺激自体淋巴细胞组为 15 6 %± 4 8%和 13 1%± 5 5 % (P <0 0 1)。结论 :U2 6 6 细胞及其可溶性抗原激发的DC与自体淋巴细胞孵育能诱导抗U2 6 6 特异性CTL。

Objective:To induce myeloma specific cytotoxic T cells response in vitro mediated by autologous dendritic cells generating from multiple myeloma(MM) patients.Methods:Monocytes isolated from the peripheral blood of MM patients were cultured in serum free medium with 800 U/ml GM CSF and 600 U/ml IFNα for 8 days to generate DCs.These DCs were pulsed by U 266 cells treated with mitomycin C and their lysates,and then incubated with autologus T lymphocytes for 5～7 days to induce antigen specific CTL.MTT assay was performed to examine the U 266 cell specific lysis.Results:DCs derived from MM patients carried a typical dendritic like morphology,they highly expressed CD86,CD54 and class II MHC molecule(HLA DR) on cell surface.Specific killing of U 266 cells mediated by DCs were tested with MTT assay.At an effector/target ratio of 20/1,specific cytotoxic activities against U 266 cells(28.0%±7.6% and 21.2%±5.4%,respectively),were observed,but without antigen stimulation,the killing rate was 11.7%±4.3%,without DC mediation,the killing rate was 15.6%±4.8% and 13.1%±5.5%(P<0.01).Conclusion:U 266 specific CTL can be induced in vitro by dendritic cells pulsed with U 266 cells and their lysates.