Optimization of Cell Infection Model for Hepatitis Delta Virus (HDV) in hNTCP-overexpressing Hepatoma Cell Lines and Analysis of HDV Genotypes

• Main Authors: Yun-Hua Lin, 林昀樺
• Other Authors: Pei-Jer Chen
• Format: Others
• Language: en_US
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/j4q5vp

碩士 === 國立臺灣大學 === 微生物學研究所 === 106 === There are 15-20 million people worldwide chronically coinfected with Hepatitis delta virus (HDV) and Hepatitis B virus (HBV). Simultaneous infection of HBV and HDV may progress easier into the most severe form of hepatitis, fulminant hepatitis, than infection of HBV only. Fulminant hepatitis causes rapid death of hepatocytes and liver failure. HDV is a satellite virus of HBV and requires HBV surface proteins (HBs Ag) to serve as viral envelopes. Since HDV is the smallest virus in animal and encodes only one protein, delta antigen, which does not possess the activity to replicate HDV genome, it must utilize host enzymes or other host factors to fulfill its life cycle. To study the role of host factors in HDV life cycle, researchers use primary human hepatocytes (PHH) and HepaRG cells in cell infection model. However, limited supply and difficult management of these cells render experiments hard to be manipulated and reproduced. The discovery of sodium-taurocholate cotransporting polypeptide (NTCP) as a HBV or HDV receptor opens a gate for researchers. Some studies indicate that overexpression of hNTCP makes cells susceptible to HBV or HDV infection. However, HDV infection rate is less than 1% even in hNTCP-overexpressing hepatoma cell line. In order to optimize the infection model in our laboratory, I used suspension infection, treatment of DMSO and PEG8000 and different clones of HDV. In our work, HDV infection rate increases from 0.1% to about 30% in the infection model. I also examine the expression of HDV genotype I, II and IV in the infection model. Since this infection model is established, the relationship between host and HDV can be further examined and eventually, find an effective antiviral therapy.