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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 the T cell epitopes of Schistosoma japonicum paramyosin. Methods The T cell epitopes of paramyosin of Schistosoma japonicum (Chinese strain) were predicted by SYFPEITHI software. The candidate epitopes were named P20, P21, P22, P23 and P24 respectively. The coding nucleotide of the candidate epitope was designed and synthesized, and cloned into the fusion expression vector pET-32c (+). The recombinant clone was identified by restriction enzyme digestion and sequencing. The positive clones were induced by IPTG and the expressed products were purified by Ni2 + -NTA column affinity chromatography and dialysis. The purified Trx fusion protein stimulated inguinal lymph node or spleen mononuclear cells in C3H / HeJ and C57BL / 6 mice in vitro, and its proliferation was detected by 3H-TdR incorporation. Results P20, P21, P22 and P23 in the candidate epitopes could effectively stimulate the proliferation of sensitized lymphocytes in C3H mice. P20 and P22 could stimulate the proliferation of sensitized lymphocytes in C57 mice. Conclusion P20 and P22 may be the universal T cell epitopes of Schistosoma japonicum paramyosin.