日本血吸虫TPI DNA疫苗基因密码子优化增强免疫保护作用的研究(英文)  

Enhancing Protective Immunity Effects of TPI DNA Vaccine against Schistosoma japonicum through Codon Optimization

作　　者：鲁飞[1] 朱荫昌[1] 戴洋[1] 王晓婷[1] 唐建霞 张纯[1] 

机构地区：[1]江苏省血吸虫病寄生虫病防治研究所、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、江苏省寄生虫病重点学科,无锡214064

出　　处：《中国寄生虫学与寄生虫病杂志》2009年第3期189-194,共6页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：Supported by UNDP/World Bank/WHO Special Program for Research and Training in Tropical Diseases (No. 991051);the Applied Basic Research Program of Health Department Fund of Jiangsu Province (No. H9918)~~

摘　　要：目的研究日本血吸虫中国大陆株磷酸丙糖异构酶基因(TPI基因)密码子优化后的DNA疫苗增强免疫保护作用的效果。方法60只BALB/c雌性小鼠随机均分为A(pcDNA3.1空质粒对照组)、B(pcDNA3.1-TPI组)、C(pcDNA3.1-TPI-mHSP70组)、D(pcDNA3.1-TPI.opt组)、E(pcDNA3.1-TPI.opt-mHSP70组)等5组。每鼠肌肉注射相应的纯化质粒DNA100μg,每隔3周免疫1次,共3次。末次免疫后4周,每鼠经腹部皮肤攻击感染日本血吸虫尾蚴(40±1)条,42d后剖杀,计数成虫及肝脏虫卵数。首次免疫前2d及感染前2d经尾静脉采血,检测IgG及IgG1、IgG2a的水平。攻击感染前2d取脾脏,制备单个脾细胞,用流式细胞仪检测白细胞介素2(IL-2)、IL-4、IL-5、γ干扰素(IFN-γ)及肿瘤坏死因子(TNF)的水平。结果B、C、D、E组小鼠血清均检测到特异性IgG及IgG2a与IgG1抗体,IgG2a/IgG1的比值分别为1.73、2.06、2.44、3.09。D、E组的IL-2、IFN-γ、TNF含量较B、C组均有不同程度地升高。D、E组减虫率分别为36.03%、39.03%,减卵率分别为41.71%、46.85%,均显著高于B、C组(P<0.01)。结论TPI基因密码子优化后的DNA疫苗相对于未优化TPIDNA疫苗能诱导小鼠产生较高的免疫保护作用,且诱导宿主产生较强的,及以Th1为主的免疫应答。Objective To study the protective effect of codon optimized TPI DNA vaccine against Schistosoma japonicum infection. Methods Sixty female BALB/c mice were randomly divided into 5 groups. The mice were injected through musculus quadriceps fexoris with 100ug pcDNA 3.1 control （Group A）, pcDNA3.1-TPI （Group B）, pcDNA 3.1-TPI-mHSP70 （Group C）, pcDNA3.1-TPI.opt （Group D）, and pcDNA3.1-TPI.opt-mHSP70 （Group E） respectively. All mice were immunized for three times with an interval of two weeks. The mice were challenged with （40±1） cercariae of S. japonicum per mouse by abdominal skin penetration 4 weeks after the last immunization, and sacrificed at 42 days post-challenge, the number of worms or hepatic eggs was counted. Blood was taken for the detection of IgG, IgG1, and IgG2a 2 days before immunization and before challenge, respectively. Spleen cells of 2 mice from each group were cultured and stimulated with ConA and rSjCTPI peptide, and the supernatant was collected for detection of IL-2, IL-4, IL-5, IFN-γ, and TNF by flow cytometry. Results ELISA showed that the mice in groups B, C, D, and E produced specific IgG and IgG1, IgG2 antibody isotypes, and the ratio of IgG2/IgG1 was 1.73, 2.06, 2.44, and 3.09, respectively. The levels of IL-2, IFN-γ and TNF in groups D and E were higher than that of groups B and C. The worm reduction rate and hepatic egg reduction rate in groups D （36.03%, 41.7%） and E （39.03%, 46.85%） were higher than those of groups B （26.28%, 28.35%） and C （28.38%, 31.39%） （P〈0.01） . Conclusions The codon optimized TPI DNA vaccine induces higher level of protective effect and Thl-biased cellular immune response than those of non-optimized TPI DNA vaccine.

关 键 词：日本血吸虫 密码子优化 免疫保护 

分 类 号：R532.21[医药卫生—内科学]