Figure 3.
pPMNs are sensitive to conventional methods of purification. (A) Human blood and oral PMNs were fixed prior to, or after, processing and labeling for flow cytometry. Density gradient centrifugation was performed and high-density (HD) PMNs were recovered and labeled for flow cytometry. Histogram plots for representative CD markers on gated PMNs are shown. Arrows indicate the pPMN population. FMO controls are shown. (B) Human blood was fixed prior to, or after, RBC lysis. A representative contour plot of CD66a × CD63 expression on gated PMNs is shown. The pPMN gate is indicated. (C) Mouse blood was fixed prior to, or after, processing and labeling for flow cytometry. Representative histograms are shown. Arrows indicate pPMN populations. FMO controls are shown. (D) Human blood was separated by density centrifugation. CD66a × CD63 scatterplots of gated PMNs are shown from a representative experiment. Representative scatterplots of whole blood PMNs, HD PMNs, low-density (LD) PMNs, and tissue PMNs (oral) PMNs are shown. (E) PMNs were enriched from whole human blood by EasySep magnetic bead negative selection. Representative contour plots are shown for whole blood (WB), bead-negative, and bead-positive populations. (F) DiffQuick images of bead-negative and bead-positive fractions indicate the presence of PMNs in both fractions. Scale bar, 10 µm.