Effect of VP2 protein of enterovirus 71 on cell injury
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摘要: 目的 研究VP2蛋白在肠道病毒 71 型(EV71)致病机制中的作用。方法 采集山东省临沂市人民医院轻症和重症手足口病(HFMD)患儿的粪便标本(SDLY 11和SDLY 107),应用MEGA 4.0软件对SDLY 11、SDLY 107 VP2 区氨基酸序列进行比对分析;参考SDLY 11、SDLY 107全序列测序结果设计 VP2 区扩增引物,构建表达载体VP2pBluescriptSK(+),通过体外转染Vero细胞表达 VP2 蛋白;乳酸脱氢酶实验检测不同临床症状病毒株的 VP2 蛋白对细胞损伤的影响。结果 序列比对分析结果显示,SDLY 11、SDLY 107 2个病毒株的 VP2 蛋白在第 199 位氨基酸处存在差异(V199I);SDLY 11、SDLY 107的 VP2 蛋白可在 Vero 细胞中表达;乳酸脱氢酶实验结果显示,2 组蛋白对细胞损伤情况差异无统计学意义(P>0.05)。结论 EV71的 VP2 蛋白可能对细胞损伤的影响不大,其在致病机制中的意义尚需进一步研究。
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关键词:
- 肠道病毒 71 型(EV71) /
- 手足口病(HFMD) /
- VP2 蛋白 /
- 细胞损伤
Abstract: Objective To study pathogenic effect of enterovirus 71(EV71)VP2 protein on cell injury.Methods SDLY 11 strain was isolated from a child patient with mild symptom of hand,foot and mouth disease(HFMD) and SDLY 107 strain was isolated from a child patient with severe symptom of HFMD.Amino acid sequences of VP2 protein were analyzed by MEGA 4 software.On the basis of genome sequences of SDLY 11 and SDLY 107,we designed the primers for the region of VP2.We built the expression vectors VP2-pBluescriptSK(+) and expressed VP2 protein in Vero cells by transfection in vitro.Lactate dehydrogenase(LDH) test was used to detect the cell injury caused by VP2 protein of diffenent EV71 strains.Results One different amino acid in VP2 protein(V199I) was found between SDLY 11 and SDLY 107.VP2 protein from SDLY 11 and SDLY 107 could be expressed in Vero cells;the results of LDH experiment showed no difference in cell injury caused by VP2 proteins of the two EV71 strains.Conclusion VP2 protein of the EV71 strains from patients with different clinical symptom may have no difference in the effect on cell injury.The result provides an important basis for the study of EV71 pathogenesis mechanism.-
Key words:
- enterovirus 71 /
- hand,foot and mouth disease /
- VP2 protein /
- cell injury
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