Intracellular Antibody Fragment Against Hepatitis B Virus X Protein Does Not Inhibit Viral Replication

다국어 초록 (Multilingual Abstract)

Replication of the hepatitis B virus is suppressed by deficiency of the X protein. Although several molecules that block cellular targets of X protein reduce the production of hepatitis B virus progeny, the effect of a specific inhibitor of X protein on viral replication has not been investigated. To block X protein specifically, we adopted an intracellular expression approach using H7 single chain variable fragment (H7scFv), an antibody fragment against X protein. We previously demonstrated that cytoplasmic expression of H7scFv inhibits X protein-induced tumorigenicity and transactivation. In this study, intracellular H7scFv expression inhibits reporter gene transactivation but not viral replication determined by endogenous hepatitis B virus polymerase activity assay and real-time PCR. Our findings imply that intracellular expression of antibody fragment against X protein may not be an alternative therapeutic modality for inhibition of hepatitis B virus replication.

참고문헌 (Reference)

1 "activation of NF-kappa B selective inhibition by antioxidants and integral membrane localization" 2991-3001, emboj1992

2 "Woodchuck hepatitis virus X protein is required for viral infection in vivo" 2026-30, jvirol1994

3 "Virus replication and virion export in X-deficient hepatitis B virus transgenic mice" 83 : 991-996, 2002

4 "The hepatitis B virus MHBst 167 protein is a pleiotropic transactivator mediating its effect via ubiquitous cellular transcription factors" 78 : 1487-1495, 1997

5 "The hepatitis B virus HBx protein is a dual specificity cytoplasmic activator of Ras and nuclear activator of transcription factors" 14 : 4747-4757, 1995

6 "The enigmatic X gene of hepatitis B virus" 78 : 12725-12734, 2004

7 "The X gene of hepatitis B virus induced growth stimulation and tumorigenic transformation of mouse NIH3T3 cells" 617-21,

8 "Real-time PCR for detection and quantitation of hepatitis B virus DNA" 65 : 250-256, 2001

9 "Oligomer synthesis by priming deficient polymerase in hepatitis B virus core particle" 322 : 22-30, 2004

10 "Intracellular single-chain antibody against hepatitis B virus core protein inhibits the replication of hepatitis B virus in cultured cells" 30 : 300-307, 1999

11 "Intracellular localization of the hepatitis B virus HBx protein" 82 : 871-882, 2001

12 "Intracellular expression of a cloned antibody fragment interferes with hepatitis B virus surface antigen secretion" 255 : 785-791, 1999

13 "Intracellular antibodies as a new class of therapeutic molecules for gene therapy" 595-601,

14 "Inhibition of cellular proteasome activities enhances hepadnavirus replication in an HBX-dependent manner" 78 : 4566-4572, 2004

15 "Identification and characterization of a structural protein of hepatitis B virus: a polymerase and surface fusion protein encoded by a spliced RNA" 275 : 398-410, 2000

16 "Hepatitis B virus X protein stimulates viral genome replication via a DDB1-dependent pathway distinct from that leading to cell death" 79 : 4238-4245, 2005

17 "Hepatitis B virus X protein is not central to the viral life cycle in vitro" 1223-7, jvirol1992

18 "Hepatitis B virus X protein colocalizes to mitochondria with a human voltage-dependent anion channel, HVDAC3, and alters its transmembrane potential" 74 : 2840-2846, 2000

19 "Hepatitis B virus HBx protein induces transcription factor AP-1 by activation of extracellular signal-regulated and c-Jun N-terminal mitogen-activated protein kinases" 70 : 4978-4985, 1996

20 "Hepatitis B virus" cancer 1988 : 61-,

21 "HBx gene of hepatitis B virus induces liver cancer in transgenic mice" 317-20, nature1991

22 "Functional neutralization of HIV-1 Vif protein by intracellular immunization inhibits reverse transcription and viral replication" 277 : 32036-32045, 2002

23 "Enhancement of hepatitis B virus replication by its X protein in transgenic mice" 76 : 2579-2584, 2002

24 "Cytosol is the prime compartment of hepatitis B virus X protein where it colocalizes with the proteasome" 16 : 2051-2063, 1998

25 "Cloning and characterization of a novel hepatitis B virus x binding protein that inhibits viral replication" 72 : 1737-1743, 1998

26 "Characterization of novel human hepatoma cell lines with stable hepatitis B virus secretion for evaluating new compounds against lamivudine- and penciclovir-resistant virus" 44 : 3402-3407, 2000

27 "Calcium signaling by HBx protein in hepatitis B virus DNA replication" 294 : 2376-2378, 2001

28 "An intracellular antibody can suppress tumorigenicity in Hepatitis B virus X-expressing cells" 55 : 569-578, 2006