Fig. 2

CD4⁺ T cell frequencies following immunization. Isolated PBMCs were stimulated with either medium alone, the vaccine antigen GMZ2, or Staphylococcal enterotoxin B (SEB) as positive control. Thereafter, intracellular cytokine staining was performed, and the cells measured by flow cytometry. Data are expressed after subtraction of unstimulated cell frequencies from that of stimulated with the positive control (SEB), and with GMZ2, and normalization with the average of positive control values. The comparison of the pro-inflammatory cytokine producing CD4⁺ T cells (Fig. 2a), and the anti-inflammatory cytokine producing CD4⁺ T cells (Fig. 2b) between D0 and D84 was performed in those receiving the control vaccine and in those vaccinated with GMZ2 (including those vaccinated with 100 µg GMZ2-Alhydrogel (opened dots), 30 µg GMZ2-CAF01 (grey dots), 100 µg GMZ2-CAF01 (black dots) using Wilcoxon test with Bonferroni correction for multiple comparisons. p value less than 0.05 is considered as statistically significant. All time points per volunteer were measured in a single experiment after several optimization tests, and individual volunteers were measured in separate experiments. Symbols represent individual samples. Red lines represent the median values with interquartile range.