[Significance of novel HBV expression vectors in selecting antiviral drugs in clinical therapy].
OBJECTIVE
To establish a new method for rapidly selecting anti-hepatitis B virus drugs in clinical therapy.


METHODS
The full-length hepatitis B virus (HBV) genomes from 8 patients with chronic hepatitis B (CHB) were generated by polymerase chain reaction (PCR). All patients were resistant to lamivudine therapy. Their HBV DNA fragments were inserted into Sap I site of pHY106 eukaryotic expression vector separately. The recombinant plasmids containing 1.1 copies of HBV genome were transfected into Huh7 cell line; the levels of HBsAg, HBeAg and HBV DNA in supernatants of Huh7 cells were measured by ELISA and real-time quantitative PCR, and intracellular HBV replicative intermediates were detected by Southern blot. Antiviral effects of lamivudine and adefovir were evaluated in this vitro system.


RESULTS
The 8 recombinant plasmids containing a full-length genome of clinical HBV isolates could replicate and be expressed in Huh 7 cells. There were 6 isolates with polymerase YVDD mutations and 2 isolates with polymerase YIDD mutations. Adefovir, but not lamivudine, inhibited the HBV replication and gene expression in vitro. Furthermore, adefovir inhibited HBV replication in these CHB patients.


CONCLUSIONS
The method described here enables a rapid selection of anti-HBV drugs in clinical therapy and is very useful in antiviral therapy for CHB patients.
Lu
Yin-ping
YP
Department of Virology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. yinpinglu@163.com
Dong
Ji-hua
JH
Liu
Zhao
Z
Guan
Shi-he
SH
Lu
Meng-ji
MJ
Yang
Dong-liang
DL
D000328:Adult; D000998:Antiviral Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D024882:Drug Resistance, Viral; D005260:Female; D006509:Hepatitis B / Q000821:virology*; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D022701:Virosomes; D055815:Young Adult
D000998:Antiviral Agents; D022701:Virosomes