日本血吸虫rSj14-3-3疫苗对小鼠抗病免疫效应的研究

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(P0.01)。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(P0.01),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。

Anti-schistosomiasis effect induced by rSj14-3-3 vaccine in BALB/c mice

Objective To investigate the anti-schistosomiasis effect in mice immunized with rSj14-3-3 vaccine.MethodsThe E.coli BL21/pET28a of rSj14-3-3 gene was coated LB/Kana/IPTG/X-gal pan,cultured,collecting bacteria,bacteria broken,separation,purification.5 μl purified protein were carried out from SDS-PAGE analysis,then to observe the results of purified protein.Each mouse in infection control group was boosted with 50 μl PBS and CFA on 0 week,and 50 μl PBS and IFA on week 2 and 4 later.While each mouce in rSj14-3-3 vaccine group was boosted with rSj14-3-3 vaccine.Two weeks after last immunization,each mouse was challenged with 30±2 cercaria of S.japonicum Chinese strain.The control group didn＇t receive the same treatment.The mice were sacrificed at the 45th day post-infection.The size of single egg granuloma in livers was measured with micrometer.The levels of hyaluronic acid（HA） and laminin（LN） in sera of the mice were determined by ELISA.ResultsThe mean hepatic egg granuloma diameters of the rSj14-3-3 vaccine group （208.5±26.3） μm ]was significantly smaller than that in infection control group （267.7±28.6） μm]（P0.01）.The levels of HA and LN in sera of rSj14-3-3 vaccine vaccinated mice were markedly lower than those in infection control group（P0.01）.ConclusionrSj14-3-3 vaccine may act as an effective inhibitor against formation of egg granuloma and reduce immunopathological damage caused by S.japonicum.