title: Modification of promoter spacer length in vaccinia virus as a strategy to control the antigen expression.
abstract: Vaccinia viruses (VACVs) with distinct early promoters have been developed to enhance antigen expression and improve antigen-specific CD8 T-cell responses. It has not been demonstrated how the length of the spacer between the coding region of the gene and its regulatory early promoter motif influences antigen expression, and whether the timing of gene expression can modify the antigen-specific CD4 T-cell response. We generated several recombinant VACVs based on the attenuated modified vaccinia Ankara (MVA) strain, which express GFP or the Leishmania LACK antigen under the control of an optimized promoter, using different spacer lengths. Longer spacer length increased GFP and LACK early expression, which correlated with an enhanced LACK-specific memory CD4 and CD8 T-cell response. These results show the importance of promoter spacer length for early antigen expression by VACV and provide alternative strategies for the design of poxvirus-based vaccines.
authors.affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
authors.forename: Mauro
authors.initials: M
authors.lastname: Di Pilato
authors.affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
authors.forename: Lucas
authors.initials: L
authors.lastname: Sánchez-Sampedro
authors.affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Mejías-Pérez
authors.affiliation: Biocomputing Unit, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
authors.forename: Carlos Oscar S
authors.initials: COS
authors.lastname: Sorzano
authors.affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
authors.forename: Mariano
authors.initials: M
authors.lastname: Esteban
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007114:Immunization; D018314:Leishmania infantum / Q000235:genetics / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000276:immunology* / Q000469:parasitology / Q000517:prevention & control; D008297:Male; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology
chemical_list: D000953:Antigens, Protozoan; D015800:Protozoan Proteins; C093139:LACK antigen, Leishmania