Phenotypical analysis of macrophages harboring different numbers of parasites.

<p>Macrophages were cultured in 96-well plates in the presence of <i>L</i>. <i>major</i> parasites (parasite/macrophage ratio 5:1). After 72 hours the cells were stained with F4/80^Cy5, CD11b^biotinylated + Streptavidin^AF546. Macrophage nuclei and parasite DNA rich areas were stained with DAPI. (A) The TissueQuest-based analysis of cells and intracellular parasites are presented. A histogram plot depicts the number of parasites within the macrophages (x-axis) and the number of macrophages harboring the indicated (see x-axis) number of <i>Leishmania</i> parasites. Cells within the highlighted gates were further analyzed regarding the expression of CD11b and F4/80. (B) The mean intensities of CD11b or F4/80 are plotted representing macrophages harboring no (#0), one (#1), six (#6) or ten (#10) parasites. Statistical analyses were performed with GraphPad Prism using the nonparametic Mann-Whitney test (p**<0,01, p*<0,05; red horizontal line represents the median). Every single dot represents one individual analyzed nucleated cell. One representative set of data out of two experiments is shown.