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目的:观察不同佐剂对不同剂量HCVDNA疫苗效果的影响.方法:雌性Balb/c小鼠分别用脂质体DDAB/EPC和氢氧化铝为佐剂的HCVDNA疫苗(由HCV结构基因、NS3和NS5b各50μg或100μg等量混合)免疫3次,ELISPOT法观察脾淋巴细胞受HCV核心、E2、E1/E2、NS3和NS5b蛋白刺激后细胞因子的产生.结果:DDAB/EPC纽产生IFN-γ和IL-4较多,用核心、E2或E1/E2刺激,显著高于裸DNA和氢氧化铝组(P<0.05).以氢氧化铝为佐剂用每种DNA100μg剂量免疫的小鼠,在受抗原刺激后,除E2外,其脾淋巴细胞产生IL-4的能力显著高于相应裸DNA组(P<0.05).在多数情况下,接受每种DNA10μg剂量的小鼠产生IFN-γ和IL-4的能力显著高于接种每种50μg者(P<0.05).与裸DNA组及DDAB/EPC组比较,氢氧化铝组小鼠淋巴细胞产生的IL-4多于IFN—γ.结论:DDAB/EPC对HCVDNA疫苗有很强的免疫佐剂效应,氢氧化铝是Th2佐剂,可将DNA疫苗Th1为主的免疫特性转换为以Th2为主,50μg的HCVDNA疫苗似不能诱导小鼠产生有效的免疫反应.
OBJECTIVE: To observe the effects of different adjuvants on the efficacy of different doses of HCVDNA vaccine.Methods: The HCVDNA vaccines adjuvanted with liposome DDAB / EPC and aluminum hydroxide were used respectively in female Balb / c mice, which consisted of HCV structural genes, NS3 and NS5b Each 50μg or 100μg equal dose) were immunized three times, ELISPOT method to observe the spleen lymphocytes by HCV core, E2, E1 / E2, NS3 and NS5b protein stimulated cytokine production.Results: DDAB / EPC New Zealand produced IFN-γ and More IL-4, stimulated with core, E2 or E1 / E2, was significantly higher than that of naked DNA and aluminum hydroxide groups (P <0.05) .Mice immunized with 100μg dose of DNA per DNA with aluminum hydroxide as adjuvant, In spite of E2 stimulation, splenic lymphocytes produced IL-4 more significantly than the corresponding naked DNA group (P <0.05), and in most cases, mice receiving doses of 10 μg per DNA produced IFN-γ and IL-4 was significantly higher than that of inoculation of 50μg each (P <0.05) .Compared with naked DNA group and DDAB / EPC group, the ALA group produced more IL-4 than IFN-γin lymphocytes of mice. : DDAB / EPC has a strong immune adjuvant effect on HCVDNA vaccine. Aluminum hydroxide is a Th2 adjuvant that can convert the immunological characteristics of DNA vaccine Th1 into predominantly Th2, 50 μg of HCVDN A vaccine does not seem to induce mice to produce an effective immune response.