利用昆蟲桿狀病毒系統表現B型肝炎核心缺損變異基因抑制B型肝炎病毒繁殖之功效(3/3)

A handy in vitro viral replication system is mandatory for hepatitis B virus (HBV) study. A recombinant baculovirus with 1.3XHBV DNA construct was previously designed to infect HepG2 cells. We adapted this system and set up another one using 1.5XHBV DNA construct to generate our recombinant baculovirus, and we use Huh7 cells instead of HepG2 cells to establish this system. HBV genome was inserted into the baculovirus by recombination and the novel HBV recombinant baculovirus was identified by enzyme digestion. The viral stock was purified and its titre was determined. We then use the HBV recombinant baculovirus to infect Huh7 cell culture and demonstrate its ability to produce HBsAg. The production of HBsAg was first detected in the media three days after infection. Its production was in proportion to the loading amount of HBV recombinant baculovirus. A sustained HBsAg production could be achieved by superinfection of this recombinant virus to the already infected Huh7 cell culture. This system can be applied to the basic and clinical studies of HBV. We then introduced the recombinant baculovirus into the portal vein of the mice to test the in vivo model. The histologic examination of the mice showed the necroinflammatory changes. However, the typical groundglass pattern of HBV infection found in human pathology was not demonstrated in the mice model. A microarray study was done to compare the RNA expression in the mouse liver infected with HBV-recombinant baculovirus and with baculovirus vector only for the future study