We constructed recombinant vaccinia viruses (VACVs) coexpressing the insulin-like growth factor-binding protein-3 (IGFBP-3) gene and the fusion gene encoding the SigE7Lamp antigen. The expression of the IGFBP-3 transgene was regulated either by the early H5 promoter or by the synthetic early/late (E/L) promoter. We have shown that IGFBP-3 expression regulated by the H5 promoter yielded higher amount of IGFBP-3 protein when compared with the E/L promoter. The immunization with P13-SigE7Lamp-H5-IGFBP-3 virus was more effective in inhibiting the growth of TC-1 tumors in mice and elicited higher T-cell response against VACV-encoded antigen than the P13-SigE7Lamp-TK(-) control virus. We found that high-level production of IGFBP-3 enhanced virus replication both in vitro and in vivo, resulting in more profound antigen stimulation. Production of IGFBP-3 was associated with a higher adsorption rate of P13-SigE7Lamp-H5-IGFBP-3 to CV-1 cells when compared with P13-SigE7Lamp-TK(-). Intracellular mature virions (IMVs) of the IGFBP-3-expressing virus P13-SigE7Lamp-H5-IGFBP-3 have two structural differences: they incorporate the IGFBP-3 protein and they have elevated phosphatidylserine (PS) exposure on outer membrane that could result in increased uptake of IMVs by macropinocytosis. The IMV PS content was measured by flow cytometry using microbeads covered with immobilized purified VACV virions.
- MeSH
- antigeny virové imunologie MeSH
- IGFBP-3 genetika imunologie MeSH
- imunizace metody MeSH
- lidský papilomavirus 16 genetika imunologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- Papillomavirus E7 - proteiny genetika imunologie MeSH
- promotorové oblasti (genetika) MeSH
- replikace viru imunologie MeSH
- T-lymfocyty imunologie MeSH
- tvorba protilátek MeSH
- vakcinace metody MeSH
- virové vakcíny imunologie farmakologie MeSH
- virus vakcinie genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Therapeutic immunization with double recombinants of vaccinia virus (VACV) co-expressing sTβRII increased rejection of established TC-1 tumors in C57BL/6 mice in comparison with single recombinant expressing SigE7LAMP. Recombinant VACV derived from vaccination strain Praha expressed either the sTβRII (ectodomain) or chimeric protein fused to immunoglobulin Fc fragment (sTβRII-Fc-Jun) under control of two different promotors together with the immunogenic tumor associated antigen HPV16 E7 oncoprotein in a form of SigE7LAMP fusion molecule. The ability of soluble receptors to bind TGF-β in vitro was proved. Immunization of mice with double recombinant viruses and virus expressing SigE7LAMP only led to eliciting similar response of E7 specific CD8+ T cells as detected by IFN-γ ELISPOT.
- MeSH
- experimentální nádory terapie MeSH
- imunizace MeSH
- lidský papilomavirus 16 imunologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- Papillomavirus E7 - proteiny imunologie MeSH
- protein-serin-threoninkinasy genetika MeSH
- receptory transformujícího růstového faktoru beta genetika MeSH
- syntetické vakcíny imunologie MeSH
- T-lymfocyty imunologie MeSH
- vakcíny proti papilomavirům imunologie MeSH
- virus vakcinie genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
