Phagocytosis capacity of CD11c⁺ cells after neuropeptide treatment.

<p>(A) Representative confocal images of <i>in vitro</i>-cultured BMDC after different treatments as indicated. Upper panel shows a merge of red fluorescence of phagocytosed <i>E. coli</i> pHrodo particles and transmitted light channel to visualize all cells. Lower panel shows only the red fluorescence according to images of the upper panel. Lower panel images were used to determine the total area of red fluorescence in each image with ImageJ software. Scale: 40 µm. (B) Graph of phagocytosis indices (PI) of BMDC and magnetically sorted total lung CD11c⁺ cells. Cells were neuropeptide or control treated for five minutes. <i>E. coli</i> pHrodo particles were then applied for 2 h. Beforehand a fraction of cells was triggered over night with LPS (three independent experiments). To inhibit phagocytosis, another fraction of cells was treated with Cytochalasin D (one experiment for BMDC, two experiments for whole lung CD11c⁺ cells). Neuropeptide-treated groups (0.1 nM VIP, 1 µM SP, 1 nM CGRP) were compared to a water-treated group (ctrl) (four independent experiments for BMDC and two independent experiments for whole lung CD11c⁺ cells). Each dot represents PI of one image. Black horizontal bars indicate the means. Per well two images were acquired. Unpaired statistical <i>t</i>-test; ns not significant; * <i>p</i><0.05; ** <i>p</i><0.01; *** <i>p</i><0.001.