Effects of G6PD Status on the Replication of Hepatitis B Virus

• Main Authors: Po Ching Chiang, 江柏慶
• Other Authors: T. Y. Chiu
• Format: Others
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/74780056328834171341

碩士 === 長庚大學 === 醫學生物技術暨檢驗學系 === 100 === Glucose-6-phosphate dehydrogenase (G6PD) plays an important role in regulating cellular redox status. G6PD deficiency is the most common emzymopathy with redox-imbalance. A preponderance of evidence show that G6PD deficiency enhances the susceptibility of cells to RNA virus infection, such as enterovirus 71, coronavirus 229E and dengue virus serotype 2. In order to investigate whether such phenomenon is also observed in DNA virus, we have used hepatitis B virus (HBV) as virus model to correlate the relationship between cellular G6PD status and HBV replication. Toward this goal, we constructed a plasmid containing 1.3-fold HBV genome called pAAV HBV1.3 which can produce HBV antigen and viral RNA. Unfortunately, a key index of viral replication-cccDNA could not be detected in the transfected cells. Hence, we choosed another plasmid named pHBV3.6 which can produce cccDNA. Once the transfection was completed, viral RNA expression was analyzed by northern blot and quantitative PCR. We found that there was no significant difference in HBV replication between control and G6PD-knockdown cells under basal condition. To further confirm our data, we measured the activity of HBV core promoter which is responsible for viral pgRNA synthesis by luciferase assay and found that G6PD status did not affect core promoter activity. In addition, using Southern blot to analyze HBV replicative intermediates, we found similar results as core promoter activity. Furthermore, there is not a significant difference in HBsAg and HBeAg content between control and G6PD-knockdown cells after pHBV3.6 transfection. In brief, we have established a system for investigating the relationship between G6PD status and HBV replication. By using this model, we found that in vitro HBV replication is not influenced by cellular G6PD activity.