Figure 4.

Polyfunctionality of CD4⁺ or CD8⁺ T cells responding to LppZ in immunized mice. Splenocytes from each mouse were collected and stimulated ex vivo with LppZ or PMA/Ionomycin. Frequencies of antigen-specific CD4⁺ (A) or CD8⁺ (C) T cells expressing single and multiple cytokines were analyzed by flow cytometry. Single or multiple cytokine production is shown on the x-axis of the bar chart (n = 3–6 mice per group). The percentages of CD4⁺ (B) and CD8⁺ (D) T cell expressing single (1+), double (2+) or triple (3+) cytokines are represented as pie charts. (E,F): frequencies of cytokine-producing CD4⁺ (E) or CD8⁺ (F) T cells upon PMA/Ionomycin stimulation *p < 0.05, **p < 0.01.