Expression, Polyubiquitination, and Therapeutic Potential of Recombinant E6E7 from HPV16 Antigens Fused to Ubiquitin
| Autor: | Liliane Maria Fernandes de Oliveira, Mirian Galliote Morale, Paulo Lee Ho, Agtha de Alencar Muniz Chaves, Marilene Demasi |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Papillomavirus E7 Proteins Recombinant Fusion Proteins Bioengineering CD8-Positive T-Lymphocytes Applied Microbiology and Biotechnology Biochemistry law.invention 03 medical and health sciences Interferon-gamma Mice Immune system Affinity chromatography Antigen Ubiquitin law Cytotoxic T cell Animals Humans Papillomavirus Vaccines Molecular Biology Human papillomavirus 16 biology Papillomavirus Infections Oncogene Proteins Viral Molecular biology Cell biology Ubiquitin ligase Mice Inbred C57BL Repressor Proteins 030104 developmental biology Gene Expression Regulation biology.protein Recombinant DNA CD8 Biotechnology |
| Zdroj: | Molecular biotechnology. 59(1) |
| ISSN: | 1559-0305 |
| Popis: | Ubiquitin-proteasome system plays an essential role in the immune response due to its involvement in the antigen generation and presentation to CD8+ T cells. Hereby, ubiquitin fused to antigens has been explored as an immunotherapeutic strategy that requires the activation of cytotoxic T lymphocytes. Here we propose to apply this ubiquitin fusion approach to a recombinant vaccine against human papillomavirus 16-infected cells. E6E7 multi-epitope antigen was fused genetically at its N- or C-terminal end to ubiquitin and expressed in Escherichia coli as inclusion bodies. The antigens were solubilized using urea and purified by nickel affinity chromatography in denatured condition. Fusion of ubiquitin to E6E7 resulted in marked polyubiquitination in vitro mainly when fused to the E6E7 N-terminal. When tested in a therapeutic scenario, the fusion of ubiquitin to E6E7 reinforced the anti-tumor protection and increased the E6/E7-specific cellular immune responses. Present results encourage the investigation of the adjuvant potential of the ubiquitin fusion to recombinant vaccines requiring CD8+ T cells. |
| Databáze: | OpenAIRE |
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