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. 2017 Apr 13;8:439. doi: 10.3389/fimmu.2017.00439

Figure 4.

Figure 4

Neutrophil extracellular traps (NETs) are a substrate to discriminate sera from microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) patients. (A–C) Naive neutrophils and (D–F) NETs form by phorbol 12-myristate 13-acetate-activated neutrophils were incubated with sera from patients with MPA (n = 8), patients with GPA (n = 8), and sera from healthy controls (Ctrl; n = 8). Left panel shows human IgG staining (green), and right panel shows overlay of human IgG (green) and DNA staining (white). Scale bars represent 50 µm. (A) Sera from MPA patients and (B) GPA patients show a cytoplasmic staining pattern in naïve neutrophils. (C) Quantification of the IgG-staining area per field of view (FOV) of naïve neutrophils. (D) Sera from MPA patients show a web-like staining pattern indicating the staining of NETs. (E) Sera from GPA patients show reactivity with the cell bodies of netting neutrophils. (F) Quantification of the IgG-staining area per FOV of NETs. p-Values in panels (C,F) were calculated using one-way-ANOVA followed by Tukey’s multiple comparison test.