Figure 5

LL37-specific T-cells are T_FH-like cells. (A) CXCR5 expression (see Fig. S4A–C) by BrdU⁺CD4⁺T-cells responding to the indicated stimuli. Results are shown as medians of percent of expression measured by flow cytometry, with Interquartile range (IQR). P values calculated by two-tailed Wilcoxon signed-rank test for intra-group comparison, and by Mann-Whitney test for inter group comparison. (B) IL-21 (pg/mL/1 × 10⁶ cells) measured by ELISA in SLE/CLE/PSO-PBMC-cultures stimulated with LL37/cit-LL37/control antigens (sample size indicated). P values by two-tailed Wilcoxon signed-rank test. (C) Cumulative data of Bcl-6/Ror-γt expression by flow cytometry of 48 hours stimulated PBMC (see Fig. S6D). Horizontal bars = means, vertical bars = standard errors of the mean, P-values by two-tailed Wilcoxon signed-rank test. (D) SLE/CLE CD4⁺ T-cell clones were cultured for three days with irradiated B-LCLs (expressing given HLA-DR) pulsed with native-LL37/cit-LL37-derived peptides (0.1-to-1 μg/mL). Percent of BrdU⁺ T-cells by flow cytometry, shown as histograms (plus/minus standard error of the mean of triplicate cultures) from one of three experiments for each T-cell clone. (E) LL37-specific T-cell clones, stimulated with PMA + Iono, were tested for cytokine expression by flow cytometry (shown dot plots of gated CD3⁺CD4⁺ cells). (F) LL37-specific T-cell clones stained for surface markers or Bcl-6/Ror-γt (intracellular staining). Expression markers (gate on CD3⁺CD4⁺ cells, black) over isotype antibody control staining (white) are shown as flow cytometry histograms. Data are from one of three representative experiments for each clone.