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 subcutaneously into mice every 4 weeks. These mice were replication-incompetent, T antigen-deleted simian virus-40 bled every 2 weeks and their sera assayed for antibody (SV40) is used as the transduction vehicle. We report here activity against HBsAg and SV40. Production of anti-HBs successful immunization using such an SV40-derived viral was measured by ELISA and confirmed by Western blot vector. Hepatitis B surface antigen (HBsAg) cDNA was analysis, both of which demonstrated significant levels of cloned downstream of two tandem SV40 early promoters anti-HBs after the second injection. We also tested proto yield a T antigen-deficient SV40 derivative, SV(HBS).duction of anti-SV40 antibodies by the ability of sera to Cultured TC7 cells were exposed to SV(HBS), and neutralize SV(HBS) infectivity. We found no evidence of expression of HBsAg was detected 24 h later by Northern neutralization of SV(HBS) infectivity even after eight inocublot and RT-PCR analysis. Immunochemistry and Western lations. Thus, replication-incompetent SV40 is itself not a blot analysis were also performed 24 h after infection to strong antigen. Our data suggest that SV40-based transdetect expression of HBsAg. Once it was ascertained that duction systems may be a useful vehicle for immunization we could express HBsAg in this way, we used SV(HBS) to and for other gene transfer applications when a need for elicit anti-HBs. SV(HBS) was injected intraperitoneally or multiple inoculations is anticipated.

SV40-derived vectors provide effective transgene expression and inhibition of HIV-1 using constitutive, conditional,and pol III promoters

“…We have shown that transduction using rSV40 vectors is equally effective in unstimulated and stimulated cells, both PBMC and CD34 + cells. 17,18 This observation was confirmed in the current study, and extended to demonstrate that SV(polyTAR) treatment of unstimulated primary human lymphoid cells can protect these cells from HIV-1 challenge. Taken together with the high transduction efficiency (Ͼ95%) 14,17 without selection, this finding suggests that rSV40, and in particular SV(polyTAR) may be an excellent candidate gene delivery vehicle for inhibiting HIV-1 in patients' blood and other lymphoid cells.…”

“…17,18 This observation was confirmed in the current study, and extended to demonstrate that SV(polyTAR) treatment of unstimulated primary human lymphoid cells can protect these cells from HIV-1 challenge. Taken together with the high transduction efficiency (Ͼ95%) 14,17 without selection, this finding suggests that rSV40, and in particular SV(polyTAR) may be an excellent candidate gene delivery vehicle for inhibiting HIV-1 in patients' blood and other lymphoid cells. Recent reports indicate that adoptive transfer of modified pbmc or T cells may be an important area of development for the control of viral infections and other diseases; 29,30 the currently reported data suggest that rSV40 gene delivery may be facilitate such therapies.…”

“…In this respect, they have delivered durable transgene expression 2,18,19 to high percentages of resting and dividing cells 1,17 without apparent immunogenicity. 17,19 Specifically, a variety of transgenes has been delivered in rSV40 vectors, directed at inhibition of HIV-1 in cultured T cell lines, 14 primary cells (D Strayer, in preparation), and in vivo in SCID-hu mice (H Goldstein et al, in preparation). The utility of these constructs reflects the combined effectiveness of potent anti-HIV-1 transgenes and efficient delivery.…”

“…SV(HBS), which we often use as a control vector in studies of cell lines, elicits immune responses in PBMC from hepatitis B-immune donors. 17 SV[CMV](polyTAR) was not used in these studies of primary cells since studies in cell lines showed that SV(polyTAR) was consistently as good as, and usually better than, SV[CMV](polyTAR) in protecting from HIV-1 challenge.…”

Exploiting hepatitis C virus activation of NFκB to deliver HCV-responsive expression of interferons α and γ

“…35 They also elicit no detectable immune response by normal animals and so can be used to deliver multiple transgenes over time and in sequence. [49][50][51] The ability of rSV40 vectors to deliver long-term transgene expression to mostly quiescent liver cells has been illustrated in murine models, in vivo: immunochemical staining carried out 7 weeks after administration of SV(BUGT) showed that X60% of hepatocytes continued to express the BUGT transgene. 6 Expression continued undiminished for X18 months.…”