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目的 :探讨肝脏Kupffer细胞 (KC)在脾内途径的肝脏靶向性基因治疗中的抗原提呈作用及可能机制。方法 :以结肠癌肝转移小鼠为模型 ,以脾内移植GM CSF基因修饰的胎肝细胞 (FLC GM)与低剂量IL 2、5 Fu腹腔注射作为基因治疗与化学免疫治疗联合应用方案。结果 :荷瘤小鼠治疗后 2w ,整个肝小叶KC数目均扩增 ;KC对99mTc 亚锡植酸钠吞噬功能明显增强 ;与KC抗原提呈功能和活化T细胞功能密切相关的共刺激分子CD40虽在对照组KC中有基础性表达 ,但在基因治疗联合化学免疫治疗组中 ,其表达水平皆有不同程度的上调 ;此外 ,联合治疗后KC体外能有效诱导同种T细胞增殖反应 ,细胞因子IL 12和IL 12的分泌水平亦升高 ,并且这些反应能被抗CD40L抗体所封闭。结论 :脾内途径的肝脏靶向性基因治疗能有效提高KC的抗原提呈功能 ,CD40 /CD40L分子在其中发挥了重要的共刺激作用
Objective : To investigate the antigen presenting effect and possible mechanism of hepatic Kupffer cells (KC) in hepatic targeting gene therapy. METHODS: In mice with liver metastasis of colon cancer as a model, intraperitoneal transplantation of GM CSF gene-modified fetal liver cells (FLC GM) and low-dose IL 2 and 5 Fu intraperitoneal injection were used as a combination therapy of gene therapy and chemotherapy immunotherapy. RESULTS: After 2 weeks of treatment, the number of KCs in the entire hepatic lobules of the tumor-bearing mice was expanded; the phagocytosis of sodium phytate in the 99mTc stannous was significantly enhanced by KC; and the co-stimulatory molecule CD40, which was closely related to the function of KC antigen presentation and the function of activated T cells. Although there is a basic expression in the control group KC, in the gene therapy combined chemotherapy immunotherapy group, the expression levels are all up-regulated to varying degrees; in addition, KC can effectively induce the same kind of T cell proliferative response in vitro in combination with cells. The secretion levels of factors IL 12 and IL 12 are also elevated, and these responses can be blocked by anti-CD40L antibodies. Conclusion : Liver-targeted gene therapy in the spleen pathway can effectively improve the antigen presentation function of KC, and CD40/CD40L molecules play an important role in co-stimulation.