日本血吸虫多价DNA疫苗免疫保护作用的研究  

Protective immunity effect of Schistosoma japonicum multivalent DNA vaccine in infected BALB/c mice

作　　者：王晓婷[1] 朱荫昌[1] 管晓虹[2] D.A.Harn 司进[1] 赵松[1] 徐明[1] 殷旭仁[1] 梁幼生[1] 何伟[1] 

机构地区：[1]江苏省血吸虫病防治研究所,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]南京医科大学分子生物学研究所 [3]美国哈佛大学公共卫生学院

出　　处：《中国血吸虫病防治杂志》2006年第1期1-5,共5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);国家高技术研究发展计划(863计划)(2002AA214181)~~

摘　　要：目的研究日本血吸虫多价DNA疫苗诱导BALB/c小鼠的免疫保护作用。方法65只BALB/c雌性小鼠随机分为5组,各组小鼠分别肌肉注射纯化的质粒DNApcDNA3.1(对照组)、pcDNA3.1-TPI(TPI)、pcDNA3.1-(CDR3)6[(CDR3)6]、pcDNA3.1-TPI-linker-(CDR3)6(TLC)和pcDNA3.1-(CDR3)6-linker-TPI(CLT),每鼠100μg。每隔2周加强免疫1次,共3次。末次免疫后4周每鼠经腹部皮肤攻击感染(45±1)条日本血吸虫尾蚴,45d后剖杀,计数成虫及肝脏虫卵数。首次免疫前2天及感染前2天经尾静脉采血检测IgG抗体水平、抗体亚类IgG1、IgG2a。末次免疫后2周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TPI、TLC组和CLT组小鼠血清都检测到特异性IgG抗体,抗体亚类IgG2a/IgG1的比值分别为1.71、2.71和4.70,而其他两组则未检测到特异性IgG抗体;TPI、TLC组和CLT组小鼠的IL-2、IFN-γ较对照组均有不同程度的升高,IL-4则无明显差异。多价DNA疫苗TLC组和CLT组减虫率分别达到37.30%、39.09%,减卵率分别为41.61%、49.54%,均显著高于TPI组和(CDR3)6组(P均<0.05)。结论多价DNA疫苗相对于单价DNA疫苗能诱导小鼠产生较高的免疫保护作用,且诱导宿主产生较高的以Th1为主的免疫应答。Objective To test the protective effect of multivalent DNA vaccine against Schistosoma japonicum infection. Methods Sixty-five female BALB/c mice of 5-6 weeks old were randomly divided into 5 groups： pcDNA3.1 group （control group）, pcDNA3.1-TPI group （TPI）, pcDNA3.1 （CDR3）6 group [（CDR3）6], pcDNA3.1-TPI-linker-（CDR3）6 group （TLC） and pcDNA3.1-（CDR3）6 linker-TPI group （CLT）. Each mouse was immunized with 100μg of pcDNA3.1 , pcDNA3.1 TPI, pcDNA3.1-（CDR3）6, pcDNA3.1-TPI-linker-（CDR3）6 or pcDNA 3.1-（CDR3） 6-1inker-TPI plasmid DNA by intramuscular injection, respectively. All animals were boosted at week 2 and week 4 with the same dosage and same method. Four weeks after the 3rd immunization, all mice were challenged with （45±1） cercariae of Schistosoma japonicum by abdominal skin penetration. Forty-five days post-challenge, the mice were sacrificed and perfused, and the numbers of recovered worms and hepatic eggs were counted. The blood was collected from the tail veins of all mice 2 days before the 1st immunization and challenge, respectively. Serum was prepared for detection of IgG, IgG1 and IgG2a. Two weeks after the 3rd immunization, the spleen cells of 2 mice from each group were euhured and stimulated with ConA and rSjCTPI peptide, and the supernatant was collected for detection of IL-2, IL-4 and IFN-γ. Results The worm reduction rates in TPI group, （CDR3）6 group, TLC group and CLT group were 26.25%, 24.10%, 37.30% and 39.09%, the hepatic egg reduction rates were 32.64%, 32.27%, 41.61% and 49.54% respectively compared with the control group. The levels of protection in TLC group and CLT group both were higher than those in TPI group and （CDR3）6 group（P〈0. 05）. ELISA results showed that the mice immunized with pcDNA3.1-TLC and pcDNA3.1-CLT produced specific IgG to rSjCTPI, while mice immunized with pcDNA3.1 and pcDNA3.1-（CDR3）6 did not. Mice in TLC group and CLT group also produced IgG1 and IgG2a antibody isotypes, with the ratios of IgG2a/Ig

关 键 词：日本血吸虫 多价DNA疫苗 免疫保护作用 

分 类 号：R383.24[医药卫生—医学寄生虫学]