Fig. 1.

Characterization of Traspain as a chimeric immunogen. Schematic representation of Traspain. Arrows point at CD8⁺ T cell epitopes included in the design with its respective mouse MHC-I haplotype (a). Immunochemical identity by Western blot. Domain-specific polyclonal antibodies (pAb) were used as primary antibody. SDS-PAGE gels were loaded as follows, lines: 1-MWM, 2-Traspain, 3-Nt-Cz (left) or ASP2 (right), arrows point at Traspain band (b). Specific antibodies response. Titers were determined by ELISA in serum samples from mice vaccinated with either Traspain or antigen combination plus c-di-AMP at 15 days post vaccination. Plates were coated with Traspain, Nt-Cz, ASP2 (left) or GELRIIKSV α-linker-peptide (right). * p < 0.05, one-way ANOVA Kruskal-Wallis test + Dunn’s multiple comparisons test (c). Proliferative response in vaccinated mice 30 days after the last dose. Spleen cells were re-stimulated with 10 µg/ml of the indicated protein. Results are expressed as proliferation index (PI), n = 6 mice per group, **p < 0.01, 2-way-ANOVA + Tukey’s multiple comparisons test (d). Neutralization assay of pAb generated upon vaccination. n = 5–6 per group, **p < 0.01, one-way-ANOVA Kruskal-Wallis test + Dunn’s multiple comparisons test (e). Results are expressed as mean ± SEM and are representative of at least three independent experiments