Figure 1
Alveolar macrophages (AMΦs) from human asthmatics have reduced cell-surface expression of CD163. (a) Confocal immunofluorescence microscopy images of bronchoalveolar lavage fluid (BALF) cells from an asthmatic subject showing CD163 expression by CD68+ AMΦs. The scale bar indicates 5 μm. (b) Gating strategy for identification of human CD163+ alveolar macrophages in BALF. Cellular debris was excluded using a forward light scatter/side scatter plot and doublets were excluded using width parameter on forward light scatter (FSC) and side light scatter (SSC) properties. CD45+ cells, that co-expressed CD14 and CD68, were identified as alveolar macrophages using side light scatter and CD45 bivariate plots from which lymphocytes had been excluded. A microscopic image of sorted CD45+/CD14+/CD68+/CD163+ cells shows a cellular population possessing typical cellular characteristics of alveolar macrophages. (c) Mean fluorescence intensity (MFI) of cell-surface CD163 expression by CD45+/CD14+/CD68+ AMΦs in BALF from normal individuals and asthmatic subjects (n=7, *P<0.008, paired t test). (d) A representative histogram overlay comparing cell-surface CD163 expression by AMΦs from a normal individual and an asthmatic subject.
