【摘要】 背景与目的:大多数多发性骨髓瘤(multiplemyeloma,MM)无法通过大剂量化疗和造血干细胞移植治愈,应用树突细胞(DCs)瘤苗清除MM患者化疗后残留的骨髓瘤细胞,是近年来骨髓瘤免疫疗法的新策略。本研究旨在探讨负载Id的DC独特型瘤苗对自体MM细胞的体外杀伤作用。方法:从MM患者外周血中分离获取DCs前体细胞,用GM鄄CSF与IL鄄4诱导分化,培养第5天加入从患者血清中提取的IgG的F(ab’)2片段(Id),第7天加TNF鄄α促成熟,将Id冲击致敏的DCs与自体T淋巴细胞共培养3天,获得肿瘤特异性细胞毒性T淋巴细胞(CTLs)。MTT法检测致敏DCs促自体T淋巴细胞增殖能力,以及患者CTLs对自体MM细胞的特异性细胞毒杀伤作用。结果:GM鄄CSF、IL鄄4和TNF鄄α联合可以有效地从MM患者外周血单核细胞中诱导出大量成熟的功能性DCs。MM患者自体血清Id冲击致敏的成熟DCs能够显著提高T细胞增殖能力,且与DC∶T的比值呈正相关;同时在1∶10时刺激细胞为负载了Id的成熟DC组刺激指数(SI)值(39.1±6.0)%,明显高于未经Id刺激的成熟DC组、经Id刺激的未成熟DC组以及未经Id刺激的未成熟DC组[(19.3±7.7)%、(15.9±6.1)%和(11.4±4.9)%]。负载了Id的成熟DC能够使幼稚T细胞活化成为肿瘤独特型CTLs,各个剂量的CTLs均能诱导出针对自体MM细胞的抑制性杀伤反应,并且?更多还原

【Abstract】 BACKGROUND & OBJECTIVE: Most multiple myeloma (MM) patients could not be cured by high-dose chemotherapy and bone marrow transplantation. This study was designed to investigate in vitro killing effect of tumor-specific cytotoxic T lymphocytes (CTLs) stimulated by idiotype protein (Id)-pulsed dendritic cells (DCs) on autologous MM cells. METHODS: DCs were generated from peripheral blood monocytes of 6 MM patients using interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF). After cultured for 5 days, immature DCs were pulsed with Id; tumor necrosis factor-α (TNF-α) was added at the 7th day. Id-pulsed DCs were cocultured with autologous T cells for 3 days to induce tumor-specific CTLs. MTT assay was used to detect proliferation of autologous T cells, and evaluate killing effect of CTLs on autologous MM cells. RESULTS: Mature DCs were successfully induced. Id-pulsed DCs markedly increased proliferation of autologous T cells in a dose-dependent manner; stimulation index (SI) of Id-pulsed DCs was the highest [(39.1±6.0)%] when the radio of DCs to T cells was 10∶1, which was significantly higher than those of unpulsed mature DCs [(19.3±7.7)%], Id-pulsed immature DCs [(15.9±6.1)%], and unpulsed immature DCs [(11.4±4.9)%] (P < 0.01). Id-pulsed DCs induced anti-MM activity of CTLs in a dose-dependent manner. Unpulsed mature DCs also induced cytotoxicity of CTLs against autologous MM cells; however, when DC∶T was 30∶1, killing rate of MM cells was significantly higher in Id-pulsed mature DCs group than in unpulsed mature DCs group [(70.1±7.9)% vs. (40.8±7.8)%,P < 0.05]. KRN7000-pulsed mature DCs stimulated proliferation of allogeneic T cells in a dose-dependent manner; when DC∶T was 1∶10, SI was significantly higher in KRN7000-pulsed mature DCs group than in unpulsed mature DCs group [(38.5±5.7)% vs. (20.2±5.7)%, P < 0.05]. CONCLUSIONS: Mature DCs could be induced and Id with biological activity could be extracted from peripheral blood of MM patients. Id-pulsed DCs could induce antitumor immune response. KRN7000 could improve the immune function of in vitro cultured DCs.更多还原