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目的初步检测抗空肠弯曲菌感染外膜黏附蛋白(PEB1)核酸疫苗与蛋白疫苗联合免疫诱导小鼠免疫应答水平。方法通过灌胃免疫的方法,将制备的核酸疫苗pcDNA3.1(-)-PEB1及蛋白疫苗PEB1采用核酸初次免疫-蛋白加强免疫及分别单独免疫的方法免疫雌性BALB/c小鼠,以PBS及空质粒pcDNA3.1(-)对照,检测各组小鼠体液免疫应答及细胞免疫应答水平。末次免疫后第28天,对各组小鼠灌胃攻击空肠弯曲菌,评价疫苗保护率。结果免疫后第56天,核酸蛋白联合免疫组小鼠脾淋巴细胞刺激指数(SI)为2.625±0.275,脾细胞培养上清中IFN-γ含量为(258.92±13.472)pg/mL。血清IgG抗体滴度为(2.507±0.124)μg/mL,胃和小肠黏膜冲洗液sIgA抗体滴度为(80.351±5.769)ng/mL,脾细胞培养上清中IL-4(377.47±14.560)pg/mL水平均明显高于各对照组(P<0.05)。小鼠攻击试验显示:核酸蛋白免疫组的疫苗保护率为91.53%。结论PEB1核酸蛋白联合免疫组诱导的免疫应答水平和疫苗保护率高于单独核酸疫苗免疫组和单独蛋白疫苗免疫组。
Objective To detect the level of immune response induced by co-immunization of Campylobacter jejuni infected with outer membrane adhesion protein (PEB1) and protein vaccine. Methods Female BALB / c mice were immunized by gavage immunization with the recombinant plasmid pcDNA3.1 (-) - PEB1 and protein vaccine PEB1 by primary immunization - protein boosting and immunization respectively. Empty plasmid pcDNA3.1 (-) control, detection of humoral immune response and cellular immune response in each group of mice. On the 28th day after the last immunization, Campylobacter jejuni was orally administered to mice in each group, and the vaccine protection rate was evaluated. Results On the 56th day after immunization, the spleen lymphocyte stimulation index (SI) of the nucleic acid protein combined immunization group was 2.625 ± 0.275, and the IFN-γ content of the spleen cell culture supernatant was (258.92 ± 13.472) pg / mL. The titer of serum IgG was (2.507 ± 0.124) μg / mL, the sIgA antibody titer of stomach and small intestinal mucosa was (80.351 ± 5.769) ng / mL and that of splenocyte culture supernatant was 377.47 ± 14.560 pg / mL levels were significantly higher than the control group (P <0.05). The mouse challenge test showed that the vaccine protection rate of the nucleic acid protein immunization group was 91.53%. Conclusion The level of immune response induced by PEB1 combined immunization group and vaccine protection rate were higher than those of immunized group and single protein vaccine alone.