Peng Hailin, Song Kai, Huang Chengyu, Ye Sai, Song Huaiguang, Hu Wei, Han Zeguang, McManus Donald P, Zhao Guoping, Zhang Qinghua
Shanghai Institute of Hematology, Ruijin Hospital, State Key Laboratory of Medical Genomics, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Exp Parasitol. 2008 May;119(1):117-24. doi: 10.1016/j.exppara.2008.01.017. Epub 2008 Feb 8.
The cDNA of a Schistosoma japonicum myophilin-like protein was cloned, sequenced, and expressed in Escherichia coli as a recombined protein (rSj myophilin-like protein), and the protein was purified by affinity chromatography. The deduced amino acid sequences of the Sj myophilin-like protein showed significant homology to myophilin, calponin, Np22 and Mp20. Northern blot and RT-PCR analyzes revealed expression of the Sj myophilin-like protein mRNA in eggs, sporocysts, cercariae, hepatic schistosomula and adult worms. Confocal fluorescence microscopy localized the native protein to the muscle of the adult worm. In schistosome-infected rabbits, the rSj myophilin-like protein antibody level, assessed by ELISA, was elevated after infection but was reduced after praziquantel treatment. In humans, the myophilin-like protein antibody level was evaluated by ELISA in sera from 33 non-infected humans and 61 schistosomiasis patients; the results showed a highly significant difference between the two groups with a sensitivity of 57.4%. Taken together, the myophilin-like protein may prove useful for monitoring the therapeutic effect of praziquantel rather than in serodiagnosis of schistosomiasis.
克隆了日本血吸虫肌动蛋白样蛋白的cDNA，进行测序，并在大肠杆菌中表达为重组蛋白（rSj肌动蛋白样蛋白），然后通过亲和层析对该蛋白进行纯化。推导的日本血吸虫肌动蛋白样蛋白氨基酸序列与肌动蛋白、钙调蛋白、Np22和Mp20具有显著同源性。Northern印迹和RT-PCR分析显示，日本血吸虫肌动蛋白样蛋白mRNA在虫卵、子孢蚴、尾蚴、肝期童虫和成虫中均有表达。共聚焦荧光显微镜检查将天然蛋白定位在成虫的肌肉中。在感染血吸虫的兔子中，通过ELISA评估，rSj肌动蛋白样蛋白抗体水平在感染后升高，但在吡喹酮治疗后降低。在人类中，通过ELISA对33名未感染人群和61名血吸虫病患者的血清中的肌动蛋白样蛋白抗体水平进行了评估；结果显示两组之间存在高度显著差异，敏感性为57.4%。综上所述，肌动蛋白样蛋白可能对监测吡喹酮的治疗效果有用，而不是用于血吸虫病的血清学诊断。
