FIGURE 1.

Immunophenotyping of monocytes. A, Gating strategy used for monocyte phenotyping. PBMCs were labelled with CD45 to show all leucocytes, then monocytes were gated from a FSC × SSC plot. Lin⁻ cells (lineage: CD3, CD19, CD20, CD56, CD66b, CD235a) were used to show the distribution of monocyte subsets by CD14 and CD16 labelling. Singlet and live/dead gating strategy plot not shown. B, The differences in septic shock severity, represented by SOFA score of D5+ survivors and early deceased patients. C, Differences in monocyte subsets depending on survival of initial phases of septic shock. There were significant differences in the frequency of classical and intermediate monocytes between D5+ survivors and early deceased patients. D and E, Differences in surface expression of HLA‐DR and CD86 in monocyte subsets. CD86 expression was significantly decreased in the classical monocytes of early deceased patients compared with survivors. F, The presence of HLA‐DR^low,neg CD86^low,neg cells in monocyte subsets is significantly different in D5+ survivors and early deceased patients. G‐I, Correlations between HLA‐DR and CD86 and sepsis severity (SOFA score) for each monocyte subset. We found the significant correlation between HLA‐DR^low,neg CD86^low,neg in classical monocyte subset with SOFA score. Data were tested by Mann‐Whitney test, error bars show SD. Correlation statistics (Spearman analysis) were performed using Graphpad software. *(P < 0.05), **(P < 0.01), ***(P < 0.001). FSC, forward scatter; HLA‐DR, human leucocyte antigen‐DR isotype; PBMCs, peripheral blood mononuclear cells; SOFA, sequential organ failure assessment; SSC, side scatter