树突细胞介导的独特型瘤苗的体外抗骨髓瘤作用

目的研究树突细胞（DC）介导的独特型瘤苗诱导骨髓瘤抗原特异性细胞毒性T淋巴细胞（CTL）的抗肿瘤免疫反应。方法从多发性骨髓瘤（MM）患者外周血中分离获取DC前体细胞，使用GM-CSF、IL-4与TNF-α诱导分化促成熟。加入MM患者的M蛋白F（ab′），片段（Id），诱导MM肿瘤特异性CTL。光学显微镜下观察其培养过程中形态特征变化，电镜观察其超微结构，间接免疫荧光观察其表型特征，采用MTT法检测致敏DC促自体T淋巴细胞增殖的能力以及患者CTL对自体骨髓瘤细胞的特异性细胞毒杀伤作用。结果 GM-CSF、IL-4和TNF-α配伍可有效地从MM患者外周血单陔细胞中诱导出大量成熟的功能性DC。MM患者自体血清Id冲击致敏的成熟DC能够显著地提高T细胞的增殖能力，并且使幼稚T细胞活化成为肿瘤独特型CTL，各个剂量的CTLs均能够产牛针对自体MM细胞的抑制性杀伤反应。结论 在MM患者外周血中可获得典型的DC，使用负载了Id的DC疫苗能够诱导出有效的抗肿瘤免疫应答反应，以DC为基础的疫苗可能在MM免疫治疗中发挥重要的作用。

In vitro anti-myeloma effects induced by myeloma idiotype-protein pulsed dendritic cell vaccine

OBJECTIVE: To investigate the specific antitumor immune response induced by idiotype protein (Id)-pulsed dendritic cells (DC) in vitro. METHODS: DC was generated from peripheral blood monocytes of the multiple myeloma (MM) patients using GM-CSF, IL-4, and TNF-alpha. The DCs were pulsed with idiotypic fragment, the F(ab')2 fragment of M protein from MM patient at the immature stage. The morphologic characteristics of the cells were observed with light and electron microscopes. The phenotypic features were analyzed with FACS, MTT assay was employed to evaluate the proliferation of autologous T cells and the inhibition rate of MM cells. RESULTS: DC precursors in peripheral blood could be induced to typical mature DC in medium containing GM-CSF, IL-4 and TNF-alpha. Mature DC with Id could increase the proliferation of the autologous T cells and activate naive T cells to become tumor specialized cytotoxic T lymphocytes (CTL). The CTL at different doses showed significant inhibition on or killing ability to autologous MM cells in vitro. CONCLUSIONS: In a suitable cytokine environment, the DC precursors from peripheral blood of MM patients could be induced to functional DC, and vaccination of Id-pulsed DC could induce active antitumor immune response.