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目的:研究CpG-ODN2216致敏的外周血单核细胞(PBMC)培养上清液对HBV相关性肝癌病人的树突状细胞(DC)成熟与功能的影响,寻求一种增强DC疫苗效果的方法。方法:从9例HBV相关性肝癌患者PBMC中诱导出未成熟的单核细胞来源的DC(MoDC),经HBV核心抗原(HBcAg)负载后,用CpG-ODN2216刺激的PBMC上清液、“细胞因子鸡尾酒(IL-1β、IL-6、TNF-α和PGE2)”以及两者的联合作用促进MoDC的进一步成熟,检测MoDC表型和功能;选择其中5例HLA-A2+病人,用成熟MoDC诱导自身T细胞产生HBV特异性CD8+的细胞毒性T淋巴细胞(CTL)。结果:用细胞因子鸡尾酒和CpG-ODN2216刺激的PBMC上清液联合作用可明显增强MoDC表面的CD80、CD83和CD1a表达,其对HBcAg负载的MoDC促成熟作用大于单独用细胞因子鸡尾酒或单独用CpG-ODN2216刺激PBMC的上清液。联合作用促进MoDC分泌IL-12和IL-10的能力明显强于单独应用PBMC上清液或细胞因子鸡尾酒,其刺激自体T细胞分泌IFN-γ、TNF-α、IL-6的能力也明显增高。联合作用促成熟的MoDC诱导HLA-A2+病人的自体T细胞产生HBVcore18-27特异性CD8+CTL的频率明显高于细胞因子鸡尾酒单独促成熟的MoDC。结论:CpG-ODN2216刺激PBMC的上清液和细胞因子鸡尾酒联合作用可以明显促进HBcAg负载的HBV相关性肝癌病人的MoDC成熟,增强MoDC分泌细胞因子、刺激自体特异性T淋巴细胞应答、诱导HBV特异性细胞毒性T细胞的能力。为提高HBV特异性树突状细胞疫苗的效果提供了一种可行方案。
OBJECTIVE: To investigate the effect of CpG-ODN2216-sensitized peripheral blood mononuclear cells (PBMCs) supernatant on the maturation and function of dendritic cells (DCs) in patients with HBV-related hepatocellular carcinoma (HCC) and to find a way to enhance the efficacy of DC vaccine . METHODS: Immature monocyte-derived DCs (MoDCs) were induced from PBMCs of 9 patients with HBV-associated HCC, and PBMC supernatants stimulated with CpG-ODN2216 after loading with HBV core antigen (HBcAg) Factor cocktail (IL-1β, IL-6, TNF-α and PGE2) and their combination to promote the further maturation of MoDC to detect the phenotype and function of MoDC. Five of the HLA-A2 + patients were selected and induced by mature MoDC Self T cells produce HBV-specific CD8 + cytotoxic T lymphocytes (CTLs). Results: The combined effect of cytokine cocktail and PBMC supernatant stimulated by CpG-ODN2216 significantly enhanced the expression of CD80, CD83 and CD1a on the surface of MoDC. The effect on the maturation of HBcAg-loaded MoDC was greater than that of cytokine cocktail or CpG alone ODN2216 stimulates PBMC supernatants. The combined effect of promoting the secretion of IL-12 and IL-10 by MoDC was significantly stronger than that of PBMC supernatant or cytokine alone, and its ability of stimulating autologous T cells to secrete IFN-γ, TNF-α and IL-6 was also significantly increased . The combined effect of mature MoDC induced autologous T cells from HLA-A2 + patients to produce HBVcore18-27-specific CD8 + CTL was significantly higher than that of cytokine cocktail alone to promote maturation of MoDC. Conclusion: CpG-ODN2216 stimulation of PBMC supernatant and cytokine cocktail can significantly promote the MoCB in patients with HBcAg-loaded HBV-related hepatocellular carcinoma, enhance the secretion of cytokines by MoDC, stimulate autologous specific T-lymphocyte response and induce HBV-specific Cytotoxic T cells. To improve the efficacy of HBV-specific dendritic cell vaccine provides a viable option.