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目的进一步了解具天然抗性东方田鼠血清识别的日本血吸虫(Schistosoma japonicum,Sj)不同发育期虫体蛋白组分特性,为保护性抗原筛选提供实验依据。方法用SDS-PAGE分离Sj不同发育期虫体蛋白,经PAS染色及考马斯亮兰染色,用Western blot法观察东方田鼠血清识别Sj不同发育期虫体的蛋白组分,根据标准分子量迁移率测算各蛋白组分分子量,比较被识别蛋白组分的特性。结果东方田鼠血清对日本血吸虫4个不同发育期虫体蛋白识别的组分数量和分子量范围不尽相同,其中对子胞蚴蛋白(SSP)识别的仅有110 kDa组分,对肺期童虫蛋白(SLWP)识别的有110kDa和45～50 kDa组分,对肝期童虫蛋白(SHWP)识别的有150、110、22和14 kDa组分,与成虫蛋白(SAWP)识别的有110、80/82、90/92、45、22和14 kDa组分。在反应强度上,以识别SLWP和SHWP中的组分为最明显。这些组分与相应蛋白电泳区带与PAS染色结果比较,4个不同发育期虫体中的110 kDa组分及SAWP中45、22和14 kDa组分为糖蛋白。结论东方田鼠血清中存在天然抗Sj多种蛋白组分的抗体,识别的主要组分存在于肺期和肝期的童虫蛋白中,并具糖蛋白特性。
Objective To further understand the characteristics of protein components of Schistosoma japonicum (Sj) at different developmental stages identified by the serum of natural voles in Microtus fortis, providing experimental evidence for the screening of protective antigens. Methods The proteins of different developmental stages of Sj were separated by SDS-PAGE and stained with PAS and Coomassie brilliant blue. Western blotting was used to observe the protein components of S. japonicus sera from different developmental stages. According to the standard molecular weight mobility The molecular weight of the protein component is compared to the identity of the identified protein component. Results The quantity and molecular weight range of the protein of Microtus ignaldis sera from four different developmental stages of Schistosoma japonicum were different. Among them, only 110 kDa was identified by SSP, 110kDa and 45-50kDa components identified by SLWP, 150,110,22 and 14 kDa components identified by SHWP and 110 identified by adult worm protein (SAWP) 80/82, 90/92, 45, 22 and 14 kDa components. In terms of reaction intensity, it is most obvious to identify the components in SLWP and SHWP. Compared with the electrophoretic zone and PAS staining, the 110 kDa components in 4 different developmental stages and the 45, 22 and 14 kDa components in SAWP were glycoproteins. Conclusion There are many antibodies against Sj protein in serum of Microtus fortis. The major components of Microtus fortis exist in the glycoproteins of lung and liver, and have the characteristics of glycoprotein.