Gating process for detecting CD14brightCD163+CD204+ cells in CD14+hPBMCs. (a) Histogram representation of CD14+ cells in peripheral blood mononuclear cells (PBMCs) by three-color staining. Upper panels show the percentage of CD14+ cells in PBMCs from (i) healthy controls and (ii) systemic sclerosis (SSc) patients. Lower panels show enlarged histogram representation of the CD14+ population in PBMCs from (iii) healthy controls and (iv) SSc patients. Values over the horizontal bar are the percentage of CD14bright cells in CD14+ population. (b) Dot plot representation of CD163+ cells in CD14+ population of PBMCs from healthy controls (left panel) and SSc patients (right panel). CD163+ cells belonged to the CD14bright subpopulation, and the percentage of CD14bright CD163+ subpopulation in SSc patients was increased compared with that in healthy controls. (c) Dot plot representation of CD163+ and CD204+ cells in CD14brightCD163+ population of PBMCs from healthy controls (left panel) and SSc patients (right panel). Further extended dot plot representation of CD14brightCD163+subpopulation (an upper right quadrant of Figure 4b) revealed that these cells co-express CD204. Data presented in a to c are representative of 16 healthy controls and 15 SSc patients. #Cells = actual number of the cells. (d) The graphic representation of quantitative result of CD14brightCD163+CD204+cells in PBMCs of SSc patients and healthy controls. Percentage of CD14brightCD163+CD204+ cells determined by flow cytometry is shown on the ordinate. P < 0.05.