Immune Unresponsiveness to Cardiopulmonary Bypass Is Associated With Unfavorable Outcome in Infants Requiring Cardiac Surgery

Figure 1. Phenotypic and Functional Alterations in Circulating Monocytes After CPB (A) Fresh peripheral blood mononuclear cells (PBMCs) were isolated from 8 infants (≤3 months of age) before (pre) and after (post) cardiopulmonary bypass (CPB). (B) Absolute counts of total leukocytes and different white blood cell types before and after CPB. (C) Absolute counts of B cells, NK cells, CD4+ T cells, and CD8+ T cells before and after CPB were determined using flow cytometry. (D) Monocytes’ subsets were characterized using flow cytometry into classical (CD14+ CD16-), nonclassical (CD14dim CD16+), and intermediate (CD14+ CD16+) monocytes. (E) Monocyte CD86 and HLA-DR expression were measured by mean fluorescence intensity (MFI) before and after CPB. (F) Representative plots gated of tumor necrosis factor (TNF)-α production by unstimulated (Unstim) and lipopolysaccharide-stimulated (LPS) monocytes before and after CPB. (G) TNF-α and interleukin (IL)-1β production by monocytes in unstimulated and stimulated monocytes before and after CPB. P values were calculated using Wilcoxon matched-pairs signed rank test. (H) Spearman’s correlation between the percentage of TNF-α and IL-1β positive monocytes…

Figure 2. Identification of a CPB-Specific Signature by Whole Blood Transcriptomic Analysis (A and B) Principal component analysis (PCA) plots of the whole blood transcriptome from 25 CPB patients (red dots) and 18 no-CPB patients (blue dots) before surgery (pre) and 6 hours after pediatric intensive care unit (PICU) admission (H6). The principal component (PC) values are presented with boxplots and were compared between CPB and no-CPB patients using Wilcoxon rank-sum tests. (C) Top genes contributing to the PC2 of the PCA performed at H6. (D and E) Comparisons of the whole blood transcriptomic profiles before and after surgery in infants from the non-CPB group and from the CPB group. Grey dots indicate genes with no significant difference in expression at H6 compared to preoperative time (adjusted P > 0.05), red and blue dots indicate genes with significantly increased and decreased expression at H6, respectively (adjusted P < 0.05). (F) UpSet plot of the number of differentially expressed genes (DEGs) at H6 compared to preoperative time, in CPB and no-CPB patients (horizontal bars), and the number of DEGs exclusively observed in CPB, no-CPB patients or shared by the whole cohort…

Figure 3. The Preoperative Inflammatory Status Affects the Outcome in CPB Patients (A) Comparison of the whole blood transcriptomic profiles before CPB, between uncomplicated and complicated patients. Grey dots indicate genes with no significant difference in expression between patient groups (adjusted P > 0.05), dark red and pink dots indicate genes with significantly increased and decreased expression in complicated CPB patients, respectively (adjusted P < 0.05). (B) Gene Set Enrichment Analysis showing the biological processes significantly depleted or enriched before surgery between the complicated and uncomplicated CPB patients (adjusted P < 0.05). Only the 40 most significant pathways are represented. (C and D) Comparisons of the plasma proteomic data before and after CPB in uncomplicated and complicated CPB patients. Grey dots indicate proteins with no significant difference in abundance (adjusted P > 0.05), red and blue dots indicate proteins with significantly increased and decreased abundance at H6, respectively (adjusted P < 0.05). (E) Unsupervised hierarchical clustering of the CPB patients based on the scaled (z-score) preoperative abundance of tumor necrosis factor…