Gene Products, tat ; genetics ; Gene Targeting ; Hepatitis B virus ; genetics ; growth & development ; Humans ; Recombinant Fusion Proteins ; genetics ; Ribonucleases ; genetics ; Transduction, Genetic ; Virus Replication ; drug effects

BACKGROUND/AIMS: The clinical outcome of patients with a partial virological response (PVR) to entecavir (ETV), in particular nucloes(t)ide analogue (NA)-experienced patients, has not been thoroughly investigated. The aim of the present study was to assess long-term outcomes in NA-naive and NA-experienced chronic hepatitis B patients with a PVR to ETV. METHODS: Chronic hepatitis B patients treated with ETV (0.5 mg/day) for at least 1 year were enrolled retrospectively. PVR was defined as a decrease in hepatitis B virus (HBV) DNA titer of more than 2 log10 IU/mL, yet with residual serum HBV DNA, as determined by real time-polymerase chain reaction, at week 48 of ETV therapy. RESULTS: A total of 202 patients (127 NA-naive and 75 NA-experienced, male 70.8%, antigen positive 53.2%, baseline serum HBV DNA 6.2 +/- 1.5 log10 IU/mL) were analyzed. Twenty-eight patients demonstrated a PVR. The PVR was associated with a high serum HBV DNA titer at baseline and at week 24. Virological response (< 60 IU/mL) was achieved in 46.2%, 61.5%, 77.6%, and 85% of patients with PVR at week 72, 96, 144, and 192, respectively. Resistance to antivirals developed in two NA-experienced patients. Failure of virological response (VR) in patients with PVR was associated with high levels of serum HBV DNA at week 48. CONCLUSIONS: Patients with PVR to ETV had favorable long-term virological outcomes. The low serum level of HBV DNA (< 200 IU/mL) at week 48 was associated with subsequent development of a VR in patients with PVR to ETV.
Adult ; Antiviral Agents/adverse effects/*therapeutic use ; Biomarkers/blood ; DNA, Viral/blood ; Drug Resistance, Viral ; Female ; Guanine/adverse effects/*analogs & derivatives/therapeutic use ; Hepatitis B virus/*drug effects/genetics/growth & development ; Hepatitis B, Chronic/diagnosis/*drug therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Time Factors ; Treatment Outcome ; Viral Load

Adult ; Cell Differentiation ; drug effects ; Cells, Cultured ; DNA, Viral ; analysis ; Female ; Fetus ; virology ; Hepatitis B virus ; genetics ; growth & development ; isolation & purification ; Hepatocytes ; cytology ; physiology ; virology ; Humans ; Nucleic Acid Hybridization ; methods ; Phenobarbital ; pharmacology ; Pregnancy ; Pregnancy Trimester, First

<b>OBJECTIVEb>To study the mechanism of signal transduction in anti-HBV action of IFN-alpha.

<b>METHODSb>The HBV DNA in HepG 2.2.15 cell line supernatant with/without IFNalpha-2b were monitored by fluorescence real-time quantitive PCR. STAT1, STAT2, ISGF3-gamma, PKR, 2'5'-OAS mRNA levels from HepG 2 and HepG 2.2.15 cell lines that were treated with/without IFNalpha-2b at different times were detected by semi-quantitive RT-PCR. And the ISGF3-gamma protein was detected by Western blot. Then, these items were detected again after inhibiting the JAK-STAT pathway with genistein.

<b>RESULTSb>The HBV DNA in 2215 supernatant that were treated with IFNalpha-2b for 8 hours decreased 0.72 log 10 copies/ml. But the basal levels of DNA in cells pretreated with genistein? followed by IFNalpha-2b did not decrease. The STAT1, STAT2, ISGF3-gamma, 2'5'-OAS, PKR mRNA levels were upregulated by IFNalpha-2b. The same phenomena were observed with STAT1, STAT2, ISGF3-gamma mRNA when pretreated with genistein then treated with IFNalpha-2b, but the levels of 2'5'-OAS, PKR mRNA were decreased in this situation. The expression of the protein of ISGF3-gamma was also augmented by IFNalpha-2b, and was blocked by genistein.

<b>CONCLUSIONb>The JAK-STAT pathway seems to be a critical pathway in IFNalpha-2b action against HBV? and ISGF3 is most probably a key factor of the route.

Antiviral Agents ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; DNA, Viral ; genetics ; isolation & purification ; Gene Expression Regulation, Neoplastic ; drug effects ; Hepatitis B virus ; drug effects ; genetics ; growth & development ; Humans ; Interferon-Stimulated Gene Factor 3 ; genetics ; metabolism ; Interferon-Stimulated Gene Factor 3, gamma Subunit ; genetics ; metabolism ; Interferon-alpha ; pharmacology ; Polymerase Chain Reaction ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; STAT1 Transcription Factor ; genetics ; metabolism ; STAT2 Transcription Factor ; genetics ; metabolism ; Signal Transduction