Expression, Polyubiquitination, and Therapeutic Potential of Recombinant E6E7 from HPV16 Antigens Fused to Ubiquitin

Author(s)

Fernandes de Oliveira, Liliane M. ; Morale, Mirian G. ; Chaves, Agtha A. M. ; Demasi, Marilene ; Ho, Paulo L.

• Document type: Journal article
• Source: MOLECULAR BIOTECHNOLOGY; v. 59, n. 1, p. 46-56, JAN 2017.
• Web of Science Citations: 0

Abstract

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. (AU)

• FAPESP's process: 10/04490-4 - Evaluation of an E6 and E7 multiepitope fused or not to ubiquitin against HPV16 derived cervical cancer
• Grantee: Liliane Maria Fernandes de Oliveira
• Support Opportunities: Scholarships in Brazil - Master