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目的:探讨腺病毒介导CD269基因修饰树突状细胞(DC)的生物学特性及体外治疗多发性骨髓瘤(MM)的效果。方法:通过基因重组技术改建一种携B细胞成熟抗原(BCMA)CD269基因的无毒性腺相关病毒,感染DC后刺激产生杀伤MM细胞的特异性细胞毒性T淋巴细胞(CTLs)。采用流式细胞仪分析CTLs的γ干扰素(IFN-γ)表达情况,采用MTS实验检测CTLs对MM原代细胞的杀伤率及效靶比。结果:CTLs IFN-γ的表达率为29.07%,明显高于对照组的13.95%(P<0.05)。MTS实验表明,培养出来的CTLs对MM细胞株U266的杀伤率达到(32.53±2.92)%,对MM病人原代细胞的杀伤率为(28.23±3.11)%,而对正常病人原代细胞的杀伤率仅为(22.11±2.54)%。随着效靶比的升高,杀伤率增高。结论:实验所构建的CD269抗原病毒能刺激培养出针对MM的特异性杀伤细胞,为临床细胞治疗奠定了有效的实验基础。
Objective: To investigate the biological characteristics of adenovirus-mediated CD269 gene-modified dendritic cells (DCs) and the therapeutic effect of multiple myeloma (MM) in vitro. Methods: A nontoxic adeno - associated virus carrying CD269 gene of B cell mature antigen (BCMA) was reconstructed by gene recombination technique. After cytotoxic DCs were induced, specific cytotoxic T lymphocytes (CTLs) that could kill MM cells were stimulated. The expression of IFN-γ in CTLs was analyzed by flow cytometry. The killing rate and target ratio of CTLs on MM primary cells were detected by MTS assay. Results: The expression of IFN-γ in CTLs was 29.07%, which was significantly higher than that in control group (13.95%, P <0.05). The results of MTS showed that the cytotoxicity of cultured CTLs to MM cell line U266 was (32.53 ± 2.92)%, and that of CTLs was (28.23 ± 3.11)%, while the killing of normal patient’s primary cells The rate was only (22.11 ± 2.54)%. With the target ratio increased, the killing rate increased. Conclusion: The CD269 antigen virus constructed in this study can stimulate the production of specific killer cells against MM, which has laid an effective experimental basis for clinical cell therapy.