FIGURE 2.

Dose-dependent efficacy of Flebogamma^® DIF against H4R3 ACPA-induced NETosis. (A) Exposure of neutrophils to H4R3 ACPA for 4 h resulted in significant increase in SYTOX^TM Green fluorescence compared to control rabbit serum at concentrations 100 ng/mL or greater. Rabbit serum alone had no significant effect. Data were fitted by non-linear fourth-parameter equation and analyzed using multiple unpaired t-tests with Benjamini, Krieger, and Yekutieli False Discovery Rate (FDR) assessment. *P < 0.05, **P < 0.01, ***P < 0.001 vs. rabbit serum at matched concentration. (B) Dose-dependency of Flebogamma^® DIF against H4R3 ACPA (100 ng/mL) induced NET formation and NETosis as quantified using SYTOX^TM Green fluorescence. Data were fitted using a non-linear fourth-parameter equation. (C) Area under the curve (AUC) of SYTOX^TM Green fluorescence over a 16 h experiment. Data were fitted using a non-linear fourth-parameter equation. Mean AUC of the Vehicle (402,513 ± 7,807) is indicated by the solid blue line, and standard deviation (S.D.) of the vehicle response is indicated by the dotted blue line.