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目的研究尿路致病性大肠埃希菌Ⅰ型菌毛重组质粒pcDNA3.0-fimH产生的免疫反应及其对小鼠的免疫保护作用。方法选取BALB/c小鼠60只,随机分为3组(PBS对照组、空质粒对照组、疫苗免疫组),3组分别用构建成功的重组质粒pcDNA3.0-fimH(为疫苗免疫组),载体质粒(pcDNA3.0)(为空质粒对照组)和PBS液(为PBS对照组),经股四头肌注射免疫小鼠,于第21天和第35天加强免疫。于免疫第0、14、28及42天收集小鼠膀胱冲洗液,检测各组SIgA水平。于免疫第42天采血,检测各组特异性IgG、IgG1、IgG2a水平。结果免疫后,疫苗免疫组小鼠膀胱灌洗液中sIgA水平随时间延长呈上升趋势,且明显高于PBS对照组及空质粒组(F=10.08,P<0.05);疫苗免疫组小鼠血清特异性IgG、IgG1、IgG2a抗体均明显高于对照组及空质粒组(P<0.05);结论 pcDNA3.0-fimH基因疫苗可诱导BALB/c小鼠产生特异性体液免疫,对小鼠尿道具有一定的免疫保护作用。
Objective To study the immune response induced by recombinant plasmid pcDNA3.0-fimH of pathogenic Escherichia coli of type Ⅰ and its protective effect on mice. Methods Sixty BALB / c mice were randomly divided into three groups (PBS control group, empty plasmid control group and vaccine group). Three groups were constructed with recombinant plasmid pcDNA3.0-fimH (vaccinated group) , PcDNA3.0 (empty plasmid control group) and PBS solution (PBS control group). The mice were immunized with quadriceps femoris and boosted on the 21st and the 35th days. Mouse bladder washings were collected on days 0, 14, 28 and 42 of immunization, and the levels of SIgA in each group were measured. Blood was collected on the 42nd day of immunization, and the levels of IgG, IgG1 and IgG2a in each group were detected. Results After immunization, the level of sIgA in the bladder lavage fluid of vaccinated mice showed an upward trend with time prolonging, which was significantly higher than that of PBS control group and empty plasmid group (F = 10.08, P <0.05) The specific IgG, IgG1 and IgG2a antibodies were significantly higher than those in control group and empty plasmid group (P <0.05). Conclusion The pcDNA3.0-fimH gene vaccine can induce specific humoral immunity in BALB / c mice, A certain degree of immune protection.