Fig. 3

Effect of Anv on fMLF- or PMA-induced degranulation by human neutrophils. Human neutrophils were incubated in the absence or the presence of different concentrations of Anv extract and then stimulated with either fMLF (a) for 5 min or PMA (b) for 15 min. Cells were pelleted (4 °C) at 8000 g for 30 s and the extracellμar medium was centrifuged at 10,000 g during 12 min before analysis by SDS-PAGE and Western blot using anti-MPO antibody. The western blots from different experiments were scanned and the intensity of MPO was quantified by densitometry analysis. The histogram shows the percentage of MPO density as compared to control values with either fMLF or PMA only. In figure c, human neutrophils were similarly incubated as above either without or with Anv extract and then stimμated with fMLF for 5 min. CD11b expression at neutrophil surface was evaluated using a PE-conjugated anti-human CD11b monoclonal antibody and flow cytometric analysis as described in Methods. Histogram shows the percentage of CD11b expression relative to fMLF response. All results are expressed as means ± SEM of three or more separate experiments. ** P < 0.01, *** P < 0.0001, compared to control