Antitumor activity and immunogenicity of recombinant vaccinia virus expressing HPV 16 E7 protein SigE7LAMP is enhanced by high-level coexpression of IGFBP-3
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
24556712
DOI
10.1038/cgt.2014.6
PII: cgt20146
Knihovny.cz E-resources
- MeSH
- Antigens, Viral immunology MeSH
- Insulin-Like Growth Factor Binding Protein 3 genetics immunology MeSH
- Immunization methods MeSH
- Human papillomavirus 16 genetics immunology MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Papillomavirus E7 Proteins genetics immunology MeSH
- Promoter Regions, Genetic MeSH
- Virus Replication immunology MeSH
- T-Lymphocytes immunology MeSH
- Antibody Formation MeSH
- Vaccination methods MeSH
- Viral Vaccines immunology pharmacology MeSH
- Vaccinia virus genetics immunology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Antigens, Viral MeSH
- Insulin-Like Growth Factor Binding Protein 3 MeSH
- oncogene protein E7, Human papillomavirus type 16 MeSH Browser
- Papillomavirus E7 Proteins MeSH
- Viral Vaccines MeSH
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.
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