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目的探讨日本血吸虫重组信号蛋白(rSj14-3-3)疫苗对小鼠的抗病免疫效应。方法将含有Sj14-3-3编码基因的E.coliBL21/pET28a涂布于LB/卡那霉素/IPTG/X-gal平板,培养、收集细菌,超声碎菌,分离、纯化,分别取5μl用SDS-PAGE法观察纯化结果,并采用BCA法测定重组蛋白的浓度。45只BALB/c小鼠随机分为3组:A组(感染对照组)于第0周每鼠经背部皮下多点注射50μl PBS及等体积完全弗氏佐剂(CFA),第2、4周每鼠皮下多点加强注射50μl PBS加等体积不完全弗氏佐剂(IFA)。B组(rSj14-3-3组)免疫方法同A组,仅将PBS改为50μg(50μl)rSj14-3-3。上述两组分别于末次免疫2周后每鼠经腹部皮肤感染日本血吸虫尾蚴(30±2)条。C组(正常对照组)未作任何处理。攻击感染45 d后剖杀小鼠,测量肝脏单个虫卵肉芽肿面积大小,并测定血清透明质酸及层黏连蛋白含量。结果 rSj14-3-3疫苗组小鼠虫卵肉芽肿直径为(208.5±26.3)μm,显著小于感染对照组的(267.7±28.6)μm(P<0.01)。rSj14-3-3疫苗组透明质酸和层黏连蛋白水平显著低于感染对照组(P<0.01),但两者均高于正常对照组。结论 rSj14-3-3疫苗具有一定的抗虫卵肉芽肿及抗肝纤维化作用。
Objective To investigate the anti-disease and immunological effects of Schistosoma japonicum recombinant signal protein (rSj14-3-3) vaccine in mice. Methods E. coli BL21 / pET28a containing Sj14-3-3 gene was coated on LB / kanamycin / IPTG / X-gal plates and cultured. The bacteria and the sonicated bacteria were separated and purified. Five microliters The purification results were observed by SDS-PAGE and the concentration of recombinant protein was determined by BCA method. Forty five BALB / c mice were randomly divided into three groups: group A (infection control group) at week 0, mice were injected subcutaneously with 50μl of PBS and an equal volume of complete Freund’s adjuvant (CFA) The rats were subcutaneously injected with 50μl PBS and an equal volume of incomplete Freund’s adjuvant (IFA). Group B (rSj14-3-3 group) immunization method with Group A, only PBS was changed to 50μg (50μl) rSj14-3-3. The above two groups were infected with cercariae of Schistosoma japonicum (30 ± 2) via abdomen skin 2 weeks after the last immunization. Group C (normal control group) did not make any treatment. Mice were sacrificed 45 days after challenge and the size of single granuloma in the liver was measured. The levels of hyaluronic acid and laminin in serum were measured. Results The diameter of the ova granuloma in mice vaccinated with rSj14-3-3 vaccine was (208.5 ± 26.3) μm, which was significantly lower than that in the infected control group (267.7 ± 28.6 μm) (P <0.01). The levels of hyaluronic acid and laminin in rSj14-3-3 vaccine group were significantly lower than those in infected control group (P <0.01), but both of them were higher than those in normal control group. Conclusion rSj14-3-3 vaccine has certain resistance to egg granuloma and anti-hepatic fibrosis.