日本血吸虫鸡尾酒式DNA疫苗与蛋白疫苗联合应用增强免疫保护作用的研究  

Enhancing protective immunity effects of the vaccine against Schistosoma japonicum infection through priming with cocktail DNA vaccines and boosting with cocktail protein vaccines

作　　者：戴洋[1] 朱荫昌[1] 王晓婷[1] 唐建霞 鲁飞[1] 徐明[1] 许永良[1] 管晓虹[2] 

机构地区：[1]江苏省血吸虫病防治研究所、卫生部寄生虫病预防与控制技术重点实验室、江苏省寄生虫分子生物学重点实验室、江苏省寄生虫病学重点学科,无锡214064 [2]南京医科大学卫生部抗体技术重点实验室

出　　处：《中国血吸虫病防治杂志》2008年第1期1-7,共7页Chinese Journal of Schistosomiasis Control

基　　金：联合国开发署/世界银行/世界卫生组织热带病研究与培训特别规划署(TDR)(991051);江苏省卫生厅应用基础基金(H9918)

摘　　要：目的探讨日本血吸虫鸡尾酒式DNA疫苗与蛋白疫苗联合应用以增强免疫保护作用的效果。方法分别大量制备质粒DNA:pcDNA3.1-SjC23、pcDNA3.1-SjCTPI、pcDNA3.1-(CDR3)6和重组蛋白SjC23-HD、SjCTPI、NP30。pcDNA3.1-SjC23、pcDNA3.1-SjCTPI、pcDNA3.1-(CDR3)6等量混合后即为鸡尾酒式的混合DNA疫苗,重组蛋白SjC23-HD、SjCTPI、NP30等量混合后即为鸡尾酒式的混合蛋白疫苗。70只BALB/c小鼠随机分为A、B、C、D、E5组,每组14只。A组为自然感染组;B组(空质粒对照组)每只小鼠分别在第0、3、6周经股四头肌注射100μlpcDNA3.1;C组(空质粒+混合蛋白对照组)每只小鼠分别在第0、3、6周经股四头肌注射100μlpcDNA3.1,第9周每鼠经背部皮下多点注射100μl混合蛋白疫苗+100μl福氏完全佐剂(FCA);D组(混合DNA组)每只小鼠分别在第0、3、6周经股四头肌注射100μl混合DNA疫苗;E组(混合DNA+混合蛋白组)每只小鼠分别在第0、3、6周经股四头肌注射100μl混合DNA疫苗,第9周每鼠经背部皮下多点注射100μl混合蛋白疫苗+100μlFCA。DNA免疫组末次免疫后4周,蛋白加强组末次免疫后2周,所有小鼠同时经腹部皮肤感染(40±1)条尾蚴。攻击感染后42d剖杀小鼠,计数成虫及肝脏虫卵数。首次免疫前2d及感染前2d分别经尾静脉采血,分离血清检测IgG抗体水平、抗体亚类IgG1及IgG2a,并取小鼠脾脏制备单个脾细胞,检测细胞因子IL-2、IL-4、IFN-γ的水平。结果C、D组和E组的减虫率分别为17.70%、32.88%和45.35%,D组和E组的减虫率均显著高于C组(P均<0.01),且E组的减虫率显著高于D组(P<0.01);C、D组和E组的减卵率分别为9.39%、36.20%和48.54%,D组和E组的减卵率均显著高于C组(P均<0.01),且E组的减虫率也显著高于D组(P<0.05)。C、D、E3组小鼠血清都检测到特异性IgG抗体,抗体亚类IgG2a/IgG1比值分别为0.525、1.829、0.712。D、E两组小鼠IL-2、IFN-γ含量较空质粒对照组均有明显升高,IL-4则无明显差异Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice. Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml, and mixed with pcDNA3.1-SjC23, pcDNA3. 1-SjCTPI, pcDNA3.1-（CDR3）6 plasmid DNAs by equal volume to form the cocktail DNA vaccine, and also mixed with recombinant proteins SjC23-HD, SjCTPI, and NP30 by equal volume to form the cocktail protein vaccine. Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups （A, B, C, D, E）. In Group A （control group）, each mouse was immunized with 100 μl saline solution by intramuscular （i. m. ） ; in Group B （pcDNA3. 1 control group）, each mouse was immunized （i. m. ） with 100 μl pcDNA3.1 for three times at week 0, 3, 6; in Group C （pcDNA3. 1 and cocktail protein group）, each mouse was immunized （i. m. ） with 100 μl pcDNA3.1 for three times at week 0, 3, 6 and immunized with 100 μl mixed protein vaccines plus 100 μl FCA by subcutaneous at week 9; in Group D （cocktail DNA vaccines group）, each mouse was immunized （i. m. ） with 100 μl mixed DNA vaccines for three times at week 0, 3, 6; in Group E （cocktail DNA vaccines plus cocktail proteins）, each mouse was immunized （i. m. ） with 100 μl mixed DNA vaccines for three times at week 0, 3, 6 and immunized with 100 μl mixed protein vaccines plus 100 μl FCA by subcutaneous at week 9. Four weeks after the last DNA immunization or two weeks after protein boosting, all the mice were challenged with （40±1） cercariae of Schistosoma japonicum by abdominal skin penetration at the same time. Forty-two days post-challenge, the mice were sacrificed and perfused, and the numbers of recovered worms and eggs in liver were counted. The blood was collected from the tail veins of all the mice two days before the first immunization and

关 键 词：日本血吸虫 混合DNA疫苗 混合蛋白疫苗 联合免疫 

分 类 号：R383.24[医药卫生—医学寄生虫学]