Xiong Yanian, Ai Dezhou, Meng Peipei, Wei Meimei, Hong Yang, Zhang Min, Huang Lini, Fu Zhiqiang, Shi Yaojun, Lin Jiaojiao
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, PR China.
Parasitol Int. 2013 Dec;62(6):522-9. doi: 10.1016/j.parint.2013.07.008. Epub 2013 Jul 25.
The schistosomal tegument is a dynamic host-interactive layer. Proteins exposed to the host on the tegumental surface are important for completion of the parasitic lifecycle. Dysferlin is a member of the ferlin family and is involved in plasma membrane repair. Based on the results of a proteomics study of tegument surface proteins of Schistosoma japonicum in our laboratory, dysferlin was identified as a tegumental protein of S. japonicum. The gene encoding S. japonicum dysferlin (SjDF), which codes for several Ca(2+) binding sites, was cloned, expressed in Escherichia coli, and characterized. Western blot analysis revealed that recombinant SjDF had good immunogenicity. Real-time RT-PCR analysis showed that SjDF was upregulated mainly in adult worms and the transcription level in 42-day-old female worms was significantly higher than that in males. Immunofluorescence analysis revealed that SjDF was mainly distributed in the tegument at various developmental stages. Experimental mice were treated with praziquantel and at 35days post-infection, we noted that damage to the tegument and subtegument worsened and did not recover at 36h post-treatment in the high-dose group and was accompanied by downregulation of SjDF mRNA, while the damage was less severe and recovered by this time in the low-dose group, and accompanied by upregulation of SjDF. Our results suggested that SjDF is a tegumental protein that may be important in schistosomal development and may participate in the repair process in muscle and tegument, and could present a viable vaccine candidate for schistosomiasis.
血吸虫的体表是一个动态的宿主交互层。体表表面暴露于宿主的蛋白质对于寄生虫生命周期的完成至关重要。肌膜蛋白是ferlin家族的成员,参与质膜修复。基于我们实验室对日本血吸虫体表表面蛋白质的蛋白质组学研究结果,肌膜蛋白被鉴定为日本血吸虫的一种体表蛋白。编码日本血吸虫肌膜蛋白(SjDF)的基因,该基因编码多个Ca(2+)结合位点,被克隆、在大肠杆菌中表达并进行了表征。蛋白质印迹分析表明重组SjDF具有良好的免疫原性。实时RT-PCR分析表明SjDF主要在成虫中上调,42日龄雌虫中的转录水平显著高于雄虫。免疫荧光分析表明SjDF主要分布在各个发育阶段的体表。用吡喹酮治疗实验小鼠,在感染后35天,我们注意到高剂量组在治疗后36小时体表和皮下组织的损伤恶化且未恢复,同时伴有SjDF mRNA的下调,而低剂量组此时损伤较轻且已恢复,并伴有SjDF的上调。我们的结果表明SjDF是一种体表蛋白,可能在血吸虫发育中起重要作用,可能参与肌肉和体表的修复过程,并且可能是血吸虫病的一个可行的疫苗候选物。