title: Use of SV40 to immunize against hepatitis B surface antigen: implications for the use of SV40 for gene transduction and its use as an immunizing agent.
abstract: We have described a novel gene transfer system, in which replication-incompetent, T antigen-deleted simian virus-40 (SV40) is used as the transduction vehicle. We report here successful immunization using such an SV40-derived viral vector. Hepatitis B surface antigen (HBsAg) cDNA was cloned downstream of two tandem SV40 early promoters to yield a T antigen-deficient SV40 derivative, SV(HBS). Cultured TC7 cells were exposed to SV(HBS), and expression of HBsAg was detected 24 h later by Northern blot and RT-PCR analysis. Immunochemistry and Western blot analysis were also performed 24 h after infection to detect expression of HBsAg. Once it was ascertained that we could express HBsAg in this way, we used SV(HBS) to elicit anti-HBs. SV(HBS) was injected intraperitoneally or subcutaneously into mice every 4 weeks. These mice were bled every 2 weeks and their sera assayed for antibody activity against HBsAg and SV40. Production of anti-HBs was measured by ELISA and confirmed by Western blot analysis, both of which demonstrated significant levels of anti-HBs after the second injection. We also tested production of anti-SV40 antibodies by the ability of sera to neutralize SV(HBS) infectivity. We found no evidence of neutralization of SV(HBS) infectivity even after eight inoculations. Thus, replication-incompetent SV40 is itself not a strong antigen. Our data suggest that SV40-based transduction systems may be a useful vehicle for immunization and for other gene transfer applications when a need for multiple inoculations is anticipated.
authors.affiliation: Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Philadelphia, PA 19107, USA.
authors.forename: R
authors.initials: R
authors.lastname: Kondo
authors.forename: M A
authors.initials: MA
authors.lastname: Feitelson
authors.forename: D S
authors.initials: DS
authors.lastname: Strayer
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D015153:Blotting, Western; D002478:Cells, Cultured; D004279:DNA, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006509:Hepatitis B / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D006510:Hepatitis B Antibodies / Q000032:analysis; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology*; D017325:Hepatitis B Vaccines / Q000008:administration & dosage* / Q000235:genetics; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D014161:Transduction, Genetic; D019444:Vaccines, DNA
chemical_list: D000914:Antibodies, Viral; D004279:DNA, Viral; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D019444:Vaccines, DNA