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Dendrimeric peptide constructs based on a lysine core that comprises both B- and T-cell epitopes of foot-and-mouth disease virus (FMDV) have proven a successful strategy for the development of FMD vaccines. Specifically, B 2 T dendrimers displaying two copies of the major type O FMDV antigenic B-cell epitope located on the virus capsid [VP1 (140–158)], covalently linked to a heterotypic T-cell epitope from either non-structural protein 3A [3A (21–35)] or 3D [3D (56–70)], named B 2 T-3A and B 2 T-3D, respectively, elicit high levels of neutralizing antibodies (nAbs) and IFN-γ-producing cells in pigs. To assess whether the inclusion and orientation of T-3A and T-3D T-cell epitopes in a single molecule could modulate immunogenicity, dendrimers with T epitopes juxtaposed in both possible orientations, i.e., constructs B 2 TT-3A3D and B 2 TT-3D3A, were made and tested in pigs. Both dendrimers elicited high nAbs titers that broadly neutralized type O FMDVs, although B 2 TT-3D3A did not respond to boosting, and induced lower IgGs titers, in particular IgG2, than B 2 TT-3A3D. Pigs immunized with B 2,  a control dendrimer displaying two B-cell epitope copies and no T-cell epitope, gave no nABs, confirming T-3A and T-3D as T helper epitopes. The T-3D peptide was found to be an immunodominant, as it produced more IFN-γ expressing cells than T-3A in the  in vitro  recall assay. Besides, in pigs immunized with the different dendrimeric peptides, CD4 +  T-cells were the major subset contributing to IFN-γ expression upon  in vitro  recall, and depletion of CD4 +  cells from PBMCs abolished the production of this cytokine. Most CD4 + IFN-γ +  cells showed a memory (CD4 + 2E3 − ) and a multifunctional phenotype, as they expressed both IFN-γ and TNF-α, suggesting that the peptides induced a potent Th1 pro-inflammatory response. Furthermore, not only the presence, but also the orientation of T-cell epitopes influenced the T-cell response, as B 2 TT-3D3A and B 2  groups had fewer cells expressing both cytokines. These results help understand how B 2 T-type dendrimers triggers T-cell populations, highlighting their potential as next-generation FMD vaccines.

Swine T-Cells and Specific Antibodies Evoked by Peptide Dendrimers Displaying Different FMDV T-Cell Epitopes