Chemotactic activity mediated by Gcf.

<p>(A) <i>In vitro</i> chemotaxis was analyzed using transwell insert plate with 5 µm pore size. 10 µg of Gcf or GcfΔCys4 were added to the bottom chamber and 5×10⁵ THP-1 cells were added to the top chamber of the plate. Media alone or media with 10% FBS was added as negative or positive control, respectively. The assembled plates were incubated at 37°C for 5 h. Cells migrated to the bottom chamber were collected and counted at least 3 times. Data are expressed as mean ± SEM percentages of migrated cells over negative control (media alone) from at least three independent experiments. (B) <i>In vivo</i> chemotaxis was examined by intranasal administration of Gcf or GcfΔCys4 in the absence of adjuvant and flow cytometric analysis of BAL cells collected 48 h after injection. BAL cells were pooled from at least three mice for each group and the percentages of cells were calculated after gating viable cells on FSC/SSC.