Postoperative changes in blood monocyte subsets.

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Postoperative changes in blood monocyte subsets. Postoperative changes in blood monocyte subsets. (A) Serial flow cytometry plots from a single representative patient, illustrating the quantification strategy (as outlined in the Methods section). Mononuclear cells were obtained from whole blood subjected to dextran sedimentation and discontinuous percoll gradients. The monocyte population was identified by gating on characteristic forward scatter and side scatter appearances. Within this gate, HLA-DR expressing cells were identified, and in this population staining for CD14 and CD16 defined ‘classical’ CD14++CD16− monocytes (Q3, bottom right quadrant), ‘intermediate’ CD14++CD16+ monocytes (Q2, top right quadrant) and ‘non-classical’ CD14+CD16++ monocytes (Q1, top left quadrant). As an additional quality control, expression of CCR2 and CX3CR1 was assessed, to confirm that the expected expression pattern of these receptors within each monocyte subset was observed. (B) Serial monocyte subset counts in blood. Median values are shown with the IQR in brackets. Paired t tests were used to determine postoperative change, n=39 preoperatively, n=38 at 24 h and n=27 at 48 h. Richard O Jones et al. BMJ Open Resp Res 2014;1:e000045 ©2014 by British Thoracic Society