DNA vaccines expressing B and T cell epitopes can protect mice from FMDV infection in the absence of specific humoral responses.
Despite foot-and-mouth disease virus (FMDV) being responsible for one of the most devastating animal diseases, little is known about the cellular immune mechanisms involved in protection against this virus. In this work we have studied the potential of DNA vaccines based on viral minigenes corresponding to three major B and T-cell FMDV epitopes (isolate C-S8c1) originally identified in natural hosts. The BTT epitopes [VP1 (133-156)-3A (11-40)-VP4 (20-34)] were cloned into the plasmid pCMV, either alone or fused to ubiquitin, the lysosomal targeting signal from LIMPII, a soluble version of CTLA4 or a signal peptide from the human prion protein, to analyze the effect of processing through different antigenic presentation pathways on the immunogenicity of the FMDV epitopes. As a first step in the analysis of modulation exerted by these target signals, a FMDV infection inhibition assay in Swiss outbred mice was developed and used to analyze the protection conferred by the different BTT-expressing plasmids. Only one of the 37 mice immunized with minigene-bearing plasmids developed specific neutralizing antibodies prior to FMDV challenge. As expected, this single mouse that had been immunized with the BTT tandem epitopes fused to a signal peptide (pCMV-spBTT) was protected against FMDV infection. Interestingly, nine more of the animals immunized with BTT-expressing plasmids did not show viremia at 48 h post-infection (pi), even in the absence of anti-FMDV antibodies prior to challenge. The highest protection (50%, six out of 12 mice) was observed with the plasmid expressing BTT alone, indicating that the targeting strategies used did not result in an improvement of the protection conferred by BTT epitopes. Interestingly, peptide specific CD4+ T-cells were detected for some of the BTT-protected mice. Thus, a DNA vaccine based on single FMDV B and T cell epitopes can protect mice, in the absence of specific antibodies at the time of challenge. Further work must be done to elucidate the mechanisms involved in protection and to determine the protective potential of these vaccines in natural FMDV hosts.
CISA-INIA, Valdeolmos, 28130 Madrid, Spain.
Belen
B
Borrego
Paloma
P
Fernandez-Pacheco
Llilianne
L
Ganges
Nieves
N
Domenech
Natalia
N
Fernandez-Borges
Francisco
F
Sobrino
Fernando
F
Rodríguez
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000235:genetics; D018955:CD36 Antigens / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D060908:CTLA-4 Antigen; D004195:Disease Models, Animal; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D005536:Foot-and-Mouth Disease / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000276:immunology*; D005822:Genetic Vectors; D007111:Immunity, Cellular; D051907:Lysosomal Membrane Proteins / Q000235:genetics; D051379:Mice; D009500:Neutralization Tests; D021381:Protein Transport / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D025801:Ubiquitin / Q000235:genetics; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*; D014766:Viremia
D000914:Antibodies, Viral; D015703:Antigens, CD; D000943:Antigens, Differentiation; D018955:CD36 Antigens; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C556707:Ctla4 protein, mouse; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D051907:Lysosomal Membrane Proteins; D011993:Recombinant Fusion Proteins; C489452:Scarb2 protein, mouse; D025801:Ubiquitin; D019444:Vaccines, DNA; D014765:Viral Vaccines