Lu Fei, Zhu Yin-Chang, Dai Yang, Wang Xiao-Ting, Tang Jian-Xia, Zhang Chun
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on Molecular Biology of Parasites, Jiangsu Provincial Key Subject on Parasitic Diseases, Wuxi 214064, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2009 Jun;27(3):189-94.
To study the protective effect of codon optimized TPI DNA vaccine against Schistosoma japonicum infection.
Sixty female BALB/c mice were randomly divided into 5 groups. The mice were injected through musculus quadriceps femoris with 100 microg 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-gamma, and TNF by flow cytometry.
ELISA showed that the mice in groups B, C, D, and E produced specific IgG and IgG1, IgG2a antibody isotypes, and the ratio of IgG2a/IgG1 was 1.73, 2.06, 2.44, and 3.09, respectively. The levels of IL-2, IFN-gamma 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) .
The codon optimized TPI DNA vaccine induces higher level of protective effect and Th1-biased cellular immune response than those of non-optimized TPI DNA vaccine.
研究密码子优化的TPI DNA疫苗对日本血吸虫感染的保护作用。
将60只雌性BALB/c小鼠随机分为5组。分别通过股四头肌注射100μg pcDNA 3.1对照（A组）、pcDNA3.1-TPI（B组）、pcDNA 3.1-TPI-mHSP70（C组）、pcDNA3.1-TPI.opt（D组）和pcDNA3.1-TPI.opt-mHSP70（E组）。所有小鼠均免疫3次，间隔2周。末次免疫后4周，每只小鼠经腹部皮肤感染（40±1）条日本血吸虫尾蚴，感染后42天处死，计数虫体或肝内虫卵数量。分别在免疫前2天和感染前采集血液检测IgG、IgG1和IgG2a。每组取2只小鼠的脾细胞进行培养，用ConA和rSjCTPI肽刺激，收集上清液，通过流式细胞术检测IL-2、IL-4、IL-5、IFN-γ和TNF。
ELISA显示，B、C、D、E组小鼠产生了特异性IgG、IgG1、IgG2a抗体亚型，IgG2a/IgG1比值分别为1.73、2.06、2.44和3.09。D组和E组的IL-2、IFN-γ和TNF水平高于B组和C组。D组（36.03% , 41.7%）和E组（39.03% , 46.85%）的减虫率和肝减卵率高于B组（26.28% , 28.35%）和C组（28.38% , 31.39%）（P<0.01）。
与未优化的TPI DNA疫苗相比，密码子优化的TPI DNA疫苗诱导产生更高水平的保护作用和以Th1为主的细胞免疫反应。
