Hepatitis B virus enhances transduction of human hepatocytes by SV40-based vectors.
OBJECTIVE
Chronic HBV infection, a world-wide epidemic, can lead to chronic hepatitis and eventually to cirrhosis and hepatocellular carcinoma. The liver poses obstacles for many available gene-transfer vectors. SV40-based vectors can transduce human hepatic and hematopoietic cells. We studied the effect of HBV on the transduction - efficiency of human hepatic cells by SV40 - based vectors.


METHODS
A SV40-vector carrying the luciferase gene, and wild-type SV40, were used to assess transduction efficiency of human HBV-positive and HBV-negative hepatic cells. Transduction efficiency was measured as luciferase activity or by T-antigen staining. To evaluate whether differences in transduction efficiency are due to cell recognition and/or nuclear transport, MHC-I receptors were measured by FACS analysis and SV40-DNA was extracted from the nuclei of transduced cells and quantified.


RESULTS
Two HBV-positive cell-lines, HepG2.2.2.15 and FLC4-A10II, were transduced significantly more efficiently than their parental HBV-negative cell-lines. Transient transfection of HuH-7 cells with the HBV genome also increased transduction efficiency. The level of MHC-I, the cellular receptor for SV40, was comparable in all the cell-lines studied. However, soon after infection with SV40, the nuclei of HepG2.2.2.15 contained >6-fold more SV40-DNA than HepG2.


CONCLUSIONS
HBV increases transduction by SV40-vectors. This is due to enhanced vector entry and/or transport into the nucleus. SV40-vectors appear to have a potential for gene therapy for the treatment of HBV infections.
Arad
Uri
U
Department of Hematology and Goldyne Savad Gene Therapy Institute, The Hebrew University-Hadassah Medical School and Hadassah University Hospital, Jerusalem, Israel.
Axelrod
Jonathan
J
Ben-nun-Shaul
Orly
O
Oppenheim
Ariella
A
Galun
Eithan
E
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D019281:Dimerization; D015870:Gene Expression; D005822:Genetic Vectors; D016679:Genome, Viral; D006515:Hepatitis B virus / Q000378:metabolism / Q000502:physiology*; D022781:Hepatocytes / Q000502:physiology*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D027601:Polyomavirus Infections / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011991:Receptors, Virus / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics*; D014161:Transduction, Genetic; D014162:Transfection; D014412:Tumor Virus Infections / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism
D004279:DNA, Viral; D015395:Histocompatibility Antigens Class I; D011991:Receptors, Virus; D014764:Viral Proteins