Study on apoptosis induced by Japanese encephalitis virus proteins

• Main Authors: Su-Lian Shiu, 許素連
• Other Authors: Cheng-Wen Lin
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/73507737950894540802

碩士 === 中國醫藥大學 === 醫學檢驗生物技術學系碩士班 === 95 === Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, causes severe neurological diseases with a high fatality rate. In this study, the goal of this study is to characterize JEV-induced apoptosis, to identify the dominant viral protein involved in apoptosis, and to investigate the molecular pathways of JEV protein-induced apoptosis. Flow cytometric analysis revealed apoptosis of HL-CZ and TE-671 cells infected with JEV at a MOI of 1 after 2-day infection. Propidium iodide staining, Annexin-V staining and fluorimetric assay of caspase-3 indicated that JEV proteins E, NS2B, NS3, and NS2b-NS3 protease induced apoptosis of HL-CZ and TE-671 cells, respectively. Western blotting showed that single expression of NS2B, NS3-180, and NS2BNS3 protease activated caspases 3 and 8 in HL-CZ cell. Active form of caspases 3 and 9 were detected in JEV E, NS3-180, and NS2BNS3 protease -expressing TE-671 cells, while active form of caspase 9 was only found in NS2b-NS3 protease-expression HL-CZ cell. In addition, the increase of ROS was measured in NS2B-, and NS2b-NS3 protease –expressing TE-671 cells. The decrease of MMP was measured in E –expressing HL-CZ cells and NS2b-NS3 protease-expression TE-671 cells. The increase of Ca2+. was measured in NS2b-NS3 protease –expressing HL-CZ cells. This study provides more information on the mechanism of the JEV induced apoptosis.