Fig. 6: T_REG and T_FH cells as important factors for vaccine-induced immune responses.

Cryopreserved PBMCs isolated from vaccine triple responders and non-responders were stained for surface antigens and intranuclear expression of FOXP3 ex vivo and analyzed by flow cytometry. A Frequencies of T_REG cells (CD4⁺CD127^lowCD25⁺FOXP3⁺) assessed by flow cytometry (1st study, n = 5 and n = 7 non-responders and responders, respectively, biologically independent samples, single missing values, see Source Data file, day 0 triple non-responders vs. responders p = 0.0390, day 6/7 triple non-responders vs. responders p = 0.0030). Plots show representative tSNE analyses of triple non-responders and responders with the expression density of FOXP3 highlighted (blue = low expression, red = high expression). B Frequencies of T_REG cells assessed by TSDR analysis and correlation with T_REG cells assessed by flow cytometry before, 3 and 70 days post-vaccination (1st study, n = 5 and n = 7 non-responders and responders, respectively, biologically independent samples, Spearman correlation). C Plot (left) depicting the tSNE analysis of 4 triple non-responders and 4 triple responders (gray) with manually gated T_REG cells highlighted and gated accordingly (CD3⁺CD4⁺FOXP3⁺ = purple). tSNE plots of non-responders and responders depicted in pseudo colors indicating low (blue) and high (red) cell densities 3 days post-vaccination. D Mean values of T_REG and T_FH cell frequencies at the indicated time points post-vaccination normalized to day 0 (1st and 2nd study, n = 10 non responders and n = 15 responders, biologically independent samples). Violin plots show the mean and the quartiles as dashed and dotted lines, respectively. The shape indicated the data distribution. Asterisks denote significant nominal p values as calculated by two-way ANOVA or Mixed-effects analysis for data sets with missing single values (uncorrected Fisher’s LSD, see Source Data file) comparing triple vaccine responders and non-responders at a given time point Source data are provided as a Source Data file.