Immunization with Human Papillomavirus 16 L1+E2 Chimeric Capsomers Elicits Cellular Immune Response and Antitumor Activity in a Mouse Model
Por:
Lopez-Toledo, Gabriela, Schaedlich, Lysann, Josabad Alonso-Castro, Angel, Monroy-Garcia, Alberto, Garcia-Rocha, Rosario, Guido, Miriam C., Gissmann, Lutz, Garcia-Carranca, Alejandro
Publicada:
1 jun 2016
Resumen:
Development of cervical cancer is associated with persistent infections
by high-risk human papillomavirus (HPV). Although current HPV L1-based
prophylactic vaccines prevent infection, they do not help to eliminate
prevalent infections or lesions. Our aims were (i) to generate a vaccine
combining prophylactic and therapeutic properties by producing chimeric
capsomers after fusion of the L1 protein to different fragments of E2
from HPV 16, and (ii) to evaluate their capacity to generate an
antitumoral cellular response, while conserving L1 neutralizing
epitopes. Chimeric proteins were produced in Escherichia coli and
purified by glutathione S-transferase (GST)-affinity chromatography.
Their structure was characterized using size exclusion chromatography,
sucrose gradient centrifugation, electron microscopy, and anti-L1
enzyme-linked immunosorbent assay. All chimeric proteins form capsomers
and heterogeneous aggregates. One, containing part of the
carboxy-terminal domain of E2 and its hinge region (L1 Delta+E2H/NC, aa
206-307), conserved the neutralizing epitope H16.V5. We then evaluated
the capacity of this chimeric protein to induce a cytotoxic T-cell
response against HPV 16 E2. In Cr-51 release cytotoxicity assays,
splenocytes from C57BL/6 immunized mice recognized and lysed TC-1/E2
cells, which express and present endogenously processed E2 peptides.
Moreover, this E2-specific cytotoxic response inhibited the growth of
tumors of TC-1/E2 cells in mice. Finally, we identified an epitope (aa
292-301) of E2 involved in this cytotoxic response. We conclude that the
L1 Delta+E2H/NC chimeric protein produced in bacteria can be an
effective and economically interesting candidate for a combined
prophylactic and therapeutic vaccine that could help eliminating
HPV16-positive low-grade cervical lesions and persistent viral
infections, thus preventing the development of lesions and, at the same
time, the establishment of new infections.
Filiaciones:
Lopez-Toledo, Gabriela:
Univ Nacl Autonoma Mexico, Dept Mol Biol & Biotechnol, Inst Invest Biomed, Mexico City 04510, DF, Mexico
Inst Nacl Cancerol SS, Basic Res Div, Lab Virus & Canc, Mexico City 14080, DF, Mexico
Schaedlich, Lysann:
German Canc Res Ctr, Div Genome Modificat & Carcinogenesis, Heidelberg, Germany
Josabad Alonso-Castro, Angel:
Inst Nacl Cancerol SS, Basic Res Div, Lab Virus & Canc, Mexico City 14080, DF, Mexico
Univ Guanajuato, Dept Pharm, Div Ciencias Nat & Exactas, Guanajuanto, Mexico
Monroy-Garcia, Alberto:
Univ Nacl Autonoma Mexico, Immunobiol Lab, Fac Estudios Super Zaragoza, Unidad Invest Diferenciac Celular & Canc,UMIEZ, Mexico City, DF, Mexico
IMSS, Lab Immunol & Canc, Unidad Invest Med Enfermedades Oncol, CMN SXXI, Mexico City, DF, Mexico
Garcia-Rocha, Rosario:
IMSS, Lab Immunol & Canc, Unidad Invest Med Enfermedades Oncol, CMN SXXI, Mexico City, DF, Mexico
IPN, Dept Immunol, Escuela Nacl Ciencias Biol, Mexico City 07738, DF, Mexico
Guido, Miriam C.:
Univ Nacl Autonoma Mexico, Dept Mol Biol & Biotechnol, Inst Invest Biomed, Mexico City 04510, DF, Mexico
Inst Nacl Cancerol SS, Basic Res Div, Lab Virus & Canc, Mexico City 14080, DF, Mexico
Gissmann, Lutz:
German Canc Res Ctr, Div Genome Modificat & Carcinogenesis, Heidelberg, Germany
Garcia-Carranca, Alejandro:
Univ Nacl Autonoma Mexico, Dept Mol Biol & Biotechnol, Inst Invest Biomed, Mexico City 04510, DF, Mexico
Inst Nacl Cancerol SS, Basic Res Div, Lab Virus & Canc, Mexico City 14080, DF, Mexico
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