日本血吸虫副肌球蛋白T细胞表位的预测和鉴定  

Identification of T Cell Epitopes on Paramyosin of Schistosoma japonicum

作　　者：王新军[1] 张兆松[1] 王勇[1] 吴海玮[1] 张蕾[1] 李光富[1] 季旻珺[1] 朱翔[1] 蔡晓萍[1] 刘丰[1] 苏川[1] 吴观陵[1] 

机构地区：[1]南京医科大学病原生物学系

出　　处：《热带医学杂志》2004年第6期653-656,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金资助项目(No.30271166)

摘　　要：目的鉴定日本血吸虫副肌球蛋白的T细胞表位。方法用SYFPEITHI软件预测日本血吸虫(中国大陆株)副肌球蛋白的T细胞表位,候选表位分别命名为P20、P21、P22、P23、P24。设计并合成候选表位的编码核苷酸,定向克隆入融合表达载体pET-32c(+),经酶切及测序鉴定出重组克隆。阳性克隆经IPTG诱导表达,表达产物以Ni2+-NTA柱亲和层析及透析纯化。纯化后的硫氧还蛋白(Trx)融合蛋白体外刺激C3H/HeJ及C57BL/6小鼠腹股沟淋巴结或脾脏单个核细胞,3H-TdR掺入法检测其增殖。结果候选表位中P20、P21、P22、P23能有效刺激C3H鼠致敏淋巴细胞细胞增殖,P20、P22能刺激C57鼠致敏淋巴细胞增殖。结论P20和P22可能是日本血吸虫副肌球蛋白的通用性T细胞表位。Objective To identify T cell epitopes on 97 000u paramyosin of Schistosoma japonicum(Sj97). Methods The primary structure of Sj97 was analysed using T cell epitope predictive algorithms SYFPEITHI and five epitope candidates(P20, P21, P22, P23 and P24)were obtained. The oligonucleotides of the candidate epitopes were designed, synthesized and inserted into the multiple cloning site of plasmid pET 32c(+). Followed with plasmids transformation into E.coli BL21, the positive clones containing the recombinant plasmids could express specific fusion proteins by IPTG introduction. The fusion protein was purified using Ni2+ NTA resin. The purified thioredoxin fusion proteins (epitope Trx) were incubated with primed lymph node cells or splenocytes of both C3H/HeJ and C57BL/6 mice, respectively, and the cell proliferation was determined by 3H TdR incorporation assay. Results The Trx fusion proteins of candidate epitopes were successfully prepared. P20, P21, P22 and P23 could stimulate the proliferation of primed lymphocytes from C3H mice, but only P20 and P22 could stimulate lymphocytes from C57BL/6 mice. Conclusion P20 and P22 should be components for an multi epitope vaccine againt Schistosoma japonicum.

关 键 词：日本血吸虫 副肌球蛋白 表位 

分 类 号：R383.24[医药卫生—医学寄生虫学]