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目的:日本血吸虫rSj26GST疫苗抑制小鼠肝脏虫卵肉芽肿形成的效果观察。方法:将含有rSj26GST编码基因的菌株E.coliBL21/p ET28a涂布LB/Kana/IPTG/X-gal平板,培养、收集细菌、超声碎菌、分离、纯化,分别取5μL进行SDS-PAGE,观察纯化结果,采用BCA法测定重组蛋白的浓度。45只BALB/c小鼠随机分为2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及等体积福氏完全佐剂(CFA),第2、第4周每鼠皮下多点加强注射50μLPBS加等体积福氏不完全佐剂(IFA);B组(rSj26GST组),同A组,唯将PBS改为50μg(50μL)rSj26GST。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剖杀小鼠,测量肝脏单个虫卵肉芽肿的大小,采用ELISA测定血清透明质酸及层黏连蛋白含量。结果:rSj26GST疫苗组小鼠虫卵肉芽肿的直径为(191.6±26.3)μm,显著小于感染对照组的(267.7±28.6)μm(P<0.05)。透明质酸和层黏连蛋白水平,rSj26GST疫苗组明显低于感染对照组(P<0.05),两者均高于正常对照组。结论:rSj26GST疫苗具有一定的抗虫卵肉芽肿及降低免疫病理损伤作用。
Objective: To observe the effect of Schistosoma japonicum rSj26GST vaccine on inhibiting the formation of egg granuloma in mice liver. Methods: LB / Kana / IPTG / X-gal plates were coated with strains of E. coli BL21 / p ET28a containing rSj26GST encoding gene. Bacteria and sonicated bacteria were isolated and purified. SDS- Purification results, the concentration of recombinant protein was determined by BCA method. Forty five BALB / c mice were randomly divided into two groups: group A (infection control group): at week 0, 50μL PBS and equal volume of complete Freunds adjuvant (CFA) In the fourth week, the mice were subcutaneously injected with 50 μL PBS and IFA, respectively. Group B (rSj26GST), same as group A, PBS was changed to 50 μg (50 μL) rSj26GST. 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): without any treatment. Mice were sacrificed on day 45 for challenge infection to measure the size of a single granuloma in the liver. Serum hyaluronic acid and laminin levels were measured by ELISA. Results: The diameter of ova granuloma in mice vaccinated with rSj26GST vaccine was (191.6 ± 26.3) μm, which was significantly lower than that of infected mice (267.7 ± 28.6 μm) (P <0.05). Hyaluronic acid and laminin levels, rSj26GST vaccine group was significantly lower than the infected control group (P <0.05), both were higher than the normal control group. Conclusion: The rSj26GST vaccine has certain resistance to egg granuloma and reduces the immunopathological damage.