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了解编码SAG1的重组质粒和IL 2基因佐剂在小鼠体内的免疫反应 ,评价该疫苗对弓形虫的保护作用。方法 编码SAG1的质粒和鼠源性IL 2表达载体以 1 0 0 μg的剂量免疫小鼠 ,3w后两次以相同的剂量加强免疫 ,分别以PBS和空质粒pcDNA3感染。采用ELISA法测定抗体水平、亚型、IFN γ和IL 4的含量 ,PCR及原位杂交检测感染鼠中DNA的整合及降解。所有鼠均由强毒力RH株弓形虫感染。结果 SAG1表达质粒 3次免疫后鼠体内特异IgG水平明显增高 ,IL 2表达质粒的联合使用导致IgG2a水平升高和IFN γ的产生。混合质闰注射的小鼠抗弓形虫感染的存活时间延长。结论 由SAG1DNA诱导的免疫应答因IL 2表达质粒的共同注射而增强 ,DNA疫苗和适当细胞因子的共同注射对抵抗弓形虫感染有较好的效果
To understand the immune response of recombinant plasmid encoding SAG1 and IL 2 gene adjuvant in mice and to evaluate the protective effect of the vaccine on Toxoplasma gondii. Methods SAG1 plasmid and murine IL2 expression vector were immunized at a dose of 100 μg. After 3 weeks, mice were boosted at the same dose twice and infected with PBS and empty plasmid pcDNA3 respectively. Antibody levels, subtypes, IFNγ and IL-4 levels were determined by ELISA. DNA integration and degradation in infected mice were detected by PCR and in situ hybridization. All mice were infected with the virulent RH strain Toxoplasma gondii. Results SAG1 expression plasmid was significantly increased in mice after three immunizations, and the combined use of IL 2 expression plasmid resulted in the increase of IgG2a level and IFNγ production. Mice injected with mixed substances extended their anti-toxoplasmosis survival. Conclusions The immune response induced by SAG1 DNA is enhanced by the co-injection of IL-2 expression plasmid. The co-injection of DNA vaccine and appropriate cytokines has a good effect against T. gondii infection