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Fig. 4 | BMC Immunology

Fig. 4

From: Macrophage migration inhibitory factor is involved in antineutrophil cytoplasmic antibody-mediated activation of C5a-primed neutrophils

Fig. 4

A macrophage migration inhibitory factor (MIF) antagonist inhibits antineutrophil cytoplasmic antibody (ANCA)-induced respiratory burst and degranulation in C5a-primed neutrophils. a Neutrophil respiratory burst was reflected by dihydrorhodamine-123 conversion into rhodamine-123. Neutrophils obtained from five healthy donors were pre-incubated with various concentrations of (S,R)3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1) for different time periods, primed with MIF and stimulated with ANCA. Blank, untreated neutrophils; C5a + ANCA, neutrophils primed with C5a and stimulated with ANCA; 5–5 min, neutrophils pre-incubated with 5 μM ISO-1 for 5 min and then treated with C5a and ANCA; 10–5 min, neutrophils pre-incubated with 10 μM ISO-1 for 5 min and then treated with C5a and ANCA; 10–15 min, neutrophils pre-incubated with 10 μM ISO-1 for 15 min and then treated with C5a and ANCA. MFI, mean fluorescence intensity. b Respiratory burst in neutrophils (obtained from five healthy donors) under various experimental conditions, reflected by dihydrorhodamine-123 conversion into rhodamine-123. DMSO, dimethyl sulfoxide (control); IgG, normal immunoglobulin G; ISO-1, (S,R)3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (MIF antagonist); MFI, mean fluorescence intensity; MPO-ANCA-positive IgG, patient-derived myeloperoxidase-ANCA-positive immunoglobulin G; PR3-ANCA-positive IgG, patient-derived membrane proteinase-3-ANCA-positive immunoglobulin G. c Degranulation of neutrophils (obtained from five healthy donors) under various experimental conditions, determined from measurements of lactoferrin amounts in cell supernatants. DMSO, dimethyl sulfoxide (control); IgG, normal immunoglobulin G; ISO-1, (S,R)3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (MIF antagonist); MPO-ANCA-positive IgG, patient-derived myeloperoxidase-ANCA-positive immunoglobulin G; PR3-ANCA-positive IgG, patient-derived membrane proteinase-3-ANCA-positive immunoglobulin G. Bars represent mean ± standard deviation of measurements made in 5 independent experiments

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