Figure 4
Anti-LL37 T-cells drive production of pathogenic anti-native LL37/cit-LL37 antibodies an. (A) SLE/PSO/HD PBMCs were stimulated for 7 days with the indicated antigens, and anti-native LL37(left)/anti-cit-LL37(right)-antibody reactivity tested by ELISA in supernatants. Sample size indicated. The results are presented as antibody fold increase of OD values with respect to untreated cells. (B) SLE PBMCs were stimulated twice with LL37/REV and anti-DNA antibodies analyzed by ELISA. (C) Immortalized B-LCLs from SLE/CLE bulk-cultures, obtained with native-LL37/cit-LL37, were tested for anti-LL37/anti-cit-LL37/anti-DNA antibody reactivity by ELISA. (D) SLE neutrophils were treated as indicated and stained for DNA (DAPI, blue), CD15 (red) (upper panels) or for LL37 (green) (middle panels, insets in the lower panels). Results from one representative experiment of four performed with different SLE neutrophils preparations. (E) SLE neutrophils were untreated (nt), or treated with the indicated stimuli; and released DNA was evaluated by PicoGreen assay (9). Results as relative fluorescence units (RFU) plus/minus standard error of the mean from five-to-ten independent experiments with different donors. RFU values of DNA/PicoGreen alone (Pico) reported. In all panels: Horizontal bars = means, vertical bars = standard errors of the mean, P values by Student’s t-test for paired samples (intragroup comparison)/unpaired (SLE vs HD or psoriasis cultures) samples.
