Fig 2: Circulating ILC Populations Decrease during the Course of Immune Maturation(A) Gating strategy including lineage markers (CD3, CD4, CD11c, CD14, CD19, CD34, CD303, TCR?d, TCRaß) to identify two dominant NK populations defined by CD56high(green) and CD16high (purple) and three ILC subsets: ILC1 (orange), ILC2 (red), and ILCP (light blue).(B) Principal component analysis (PCA) and heatmap shown for each replicate for each participant (see Table S1).(C) DEGs among ILC2, ILCP, CD56high (NKCD56), and CD16high (NKCD16) NK cell populations from four HIV-negative and six HIV-positive pediatric subjects.(D) Frequencies of total helper ILC subsets as defined in (A), comparing HIV-negative newborn (NB) (n = 39), pediatric (2–5 years, n = 12), pediatric (>5 years, n = 25), and adult (n = 62) individuals expressed as percentage of total CD45+ lymphocytes.(E) As in (C) but showing frequencies of total NK and subset-specific differences between pediatric and adult subjects.p values by Dunn’s multiple comparisons test.

Fig 3: Bar graph showing log fold change of CD34+ marker for each patient, identified by case number, computed at different time points with respect to three baselines: Induction, sternotomy and 0 hr.(A) Log fold change at on-pump 0hr with respect to induction. (B) Log fold change for on-pump 0hr time point with respect to sternotomy. (C) Shows 24hr log fold change with respect to sternotomy. Case numbers for patients in G1 (insignificant vasoplegia) are shown in green and those for patients in G2 (significant vasoplegia) are shown in red. The status of individual patients as diabetic (d), hypertensive (h) or having renal failure (r) is also indicated together with each bar. (D), (E), (F) show machine learning groupings for G1 and G2 using logistic regression and (G), (H), (I) show the same based on gradient boosting. Correctly classified patients are shown, by case number, in green boxes, and misclassified patients in red boxes. Corresponding p-values are shown.

Fig 4: Circulating counts of progenitor cells in individuals undergoing CPB at various intra-operative time points in G1 and G2 study individuals.Line graph showing fold change in cell counts for (A) CD34+ (B) HSC (CD34+CD133+) and (C) EPC (CD34+VEGFR2+) in vasoplegic groups namely G1 and G2. Data is plotted as mean ± SEM. *p = 0.05, ** p = 0.01 vs G2 at that point. Mann-Whitney’s test and Wilcoxon matched pair test were employed accordingly.

Fig 5: Identification of a CD19–IL-7R+ Progenitor(A) Fluorescence-activated cell sorting (FACS) analysis of CS17 FL showing CD19-IL-7R+ progenitor co-expressing CD34 and CD45RA. Adult BM (right) is shown for comparison. Cells were gated CD45+LIN-CD19–. Further gating as indicated. Mean percentage of total CD45+CD34+ cells.(B) Kinetics of IL-7R progenitor emergence during development as percentage of total CD45+CD34+ cells. Each dot represents one biological sample. Mean with range. Statistical comparison versus CB control.(C) B cell potential of IL-7R+KIT+ FL progenitors was tested by co-culture on MS-5 for 2 weeks. Percentage of CD19+ B cells of total CD45+ cells is shown (left) with representative FACS analysis (right). CD34+ cells were used as control. Mean ± SD, n = 2–3.(D) Agarose gel of IGH DJ rearrangements in IL-7R+ FL progenitor. Top: DH7JH recombination assay. Bottom: DH1-6JH recombination assay. Label for lanes 1–6 is shown to the right. Marker lanes (M). Predicted DH7JH recombination between 100 and 130 bp. Predicted DH1-6JH rearrangements between 110 and 290 bp, n = 1 (van Dongen et al., 2003).(E) GM and erythroid potential of IL-7R+KIT+ FL progenitor in semi-solid media. Data show distribution of colony types as percentage of plated cells versus CD34+ cells as controls. Mean ± SD, n = 2 per developmental stage.(F) Myeloid potential of IL-7R+KIT+ FL progenitor in liquid culture (20–40 cells/well). Graph shows percentage of wells expanded to >50 cells (left). Cytospin of macrophages generated from FL IL-7R+KIT+ progenitor (right). Mean ± SD, n = 3.See also Figure S2.