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背景与目的:Fms样酪氨酸激酶3配体(fms-1ike tyrosine kinase 3 ligand,FLT3-L)、CpG虽可作为免疫调节剂发挥抗肿瘤作用,但单独用于治疗肿瘤效果不佳。本研究旨在探讨FLT3-L+肿瘤抗原+CpG联合应用对肝癌小鼠的治疗效果及机制,并探讨FLT3-L与GM-CSF在肿瘤治疗效果方面的差异。方法:选择小鼠H22肝癌模型为研究对象,分为FLT3-L+CpG组、GM-CSF+CpG组和PBS组。皮下注射给药后,用流式细胞术检测各组小鼠体内免疫细胞激活情况,免疫组化法检测肿瘤组织及局部细胞浸润情况,ELISA法检测小鼠体液免疫激活情况,同时观察各组小鼠生存期的差异,评价各组药物的抗肿瘤效果及可能的作用机制。结果:(1)FLT3-L+CpG组小鼠肿瘤生长明显被抑制,小鼠生存期显著延长,同GM-CSF+CpG组和PBS组相比差异具有统计学意义(P<0.05)。(2)脾脏淋巴细胞亚群分析显示,FLT3-L+CpG组CD3~+T细胞比例显著上升,CD4~+、CD8~+亚群比例较PBS组与GM-CSF+CpG组均明显上升,CD4~+CD25~+Treg细胞比例明显下降,差异有统计学意义(P<0.05)。(3)肿瘤组织形态学分析显示,FLT3-L+CpG组可见大片的组织坏死和显著的淋巴细胞浸润。(4)肿瘤局部浸润淋巴细胞表型分析显示,FLT3-L+CpG组肿瘤局部NK细胞、DC细胞、CD8~+T细胞明显增加,较各治疗组均有明显变化,与PBS相比差异有统计学意义(P<0.05)。(5)FLT3-L+CDG组与GM-CSF+CpG组小鼠体内细胞因子IFN-γ、TNF-α、IL-2、IL-12均明显升高(P<0.05)。结论:FLT3-L+肿瘤抗原+CpG联合应用可发挥明显的抗肿瘤作用。此方案可能通过打破肿瘤免疫耐受,激活和增加循环中T细胞,并趋化NK细胞、DC细胞、CD8~+T细胞到肿瘤局部,发挥杀伤肿瘤细胞的作用:同时可活化免疫因子网络,通过细胞免疫和体液免疫共同发挥抗肿瘤作用。
BACKGROUND & OBJECTIVE: Fms-1-kinase tyrosine kinase 3 ligand (FLT3-L) and CpG can exert anti-tumor effects as immunomodulators, but are not effective in the treatment of tumors alone. The purpose of this study was to investigate the therapeutic effect and mechanism of FLT3-L + tumor antigen combined with CpG on mice with hepatocellular carcinoma and to explore the differences between FLT3-L and GM-CSF in tumor therapy. Methods: The mouse H22 hepatocellular carcinoma model was selected as the study object and divided into FLT3-L + CpG group, GM-CSF + CpG group and PBS group. Subcutaneous injection after administration, the activation of immune cells in each group was detected by flow cytometry, the tumor tissue and local cell infiltration were detected by immunohistochemistry, and the humoral immunity activation was detected by ELISA. At the same time, The difference of survival time of rats was evaluated. The anti-tumor effect and possible mechanism of each drug were evaluated. Results: (1) The growth of mice in FLT3-L + CpG group was significantly inhibited and the survival time of mice was significantly prolonged. The difference was statistically significant compared with that of GM-CSF + CpG group and PBS group (P <0.05). (2) Spleen lymphocyte subsets analysis showed that the proportion of CD3 ~ + T cells in FLT3-L + CpG group increased significantly, and the proportion of CD4 ~ + and CD8 ~ + subsets in FLT3-L + CpG group increased significantly compared with PBS group and GM- The proportion of CD4 ~ + CD25 ~ + Treg cells was significantly decreased, the difference was statistically significant (P <0.05). (3) Morphological analysis of tumor showed that a large number of tissue necrosis and significant lymphocyte infiltration were observed in FLT3-L + CpG group. (4) Phenotypic analysis of local infiltrating lymphocytes showed that the number of local NK cells, DCs and CD8 + T cells in FLT3-L + CpG group were significantly increased compared with those in each treatment group Statistical significance (P <0.05). (5) The levels of cytokines IFN-γ, TNF-α, IL-2 and IL-12 in FLT3-L + CDG group and GM-CSF + CpG group were significantly increased (P <0.05). Conclusion: The combined application of FLT3-L + tumor antigen and CpG can exert obvious anti-tumor effect. This program may play a role in killing tumor cells by breaking the tumor immune tolerance, activating and increasing circulating T cells, and chemotaxis of NK cells, DC cells and CD8 + T cells to the tumor site. At the same time, it can activate the immune factor network, Antitumor effect by both cellular and humoral immunity.