Fig. 6.

Assessment of specific CD8⁺ T-cell responses elicited after i.m. immunization with PyCSP-derived peptide mixed with 7DW8-5 and MPLA. Splenocytes were obtained from BALB/c mice after three doses of i.m. immunization with a PyCSP peptide alone, or with a PyCSP peptide mixed with 7DW8-5 and/or MPLA, and the tetramer assay was performed. Splenocytes from a group of non-immunized mice (n = 5) were used as a negative control. In (A), flow cytometric plots of CD8 versus SYVPSAEQI-loaded tetramer are shown for all immunization schemes as well as non-immunized controls after selection of naïve (CD44⁻CD62L^+/−), central (CD44⁺CD62L⁺) and effector (CD44⁺CD62L⁻) subsets as described. In (B), the percentage of SYVPSAEQI-loaded tetramer⁺ T-cells among naïve (CD44⁻CD62L^+/−), central (CD44⁺CD62L⁺) and effector (CD44⁺CD62L⁻) CD8⁺ T-cells subsets, respectively was measured by FACS. The cell subsets were indicated by the colors (naïve –white; central – gray; effector – black). In this figure, experiments were repeated twice and in duplicates. Statistical significance was displayed as ***, **, or *, if the p value is <0.001, <0.01, or <0.05, respectively. The abbreviation - n.s. - stands for “not significant” and was assigned if p ≥ 0.05.