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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing <t>flow-cytometry</t> data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.
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Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing flow-cytometry data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.

Journal: bioRxiv

Article Title: A 3-genes interferon signature predicts sustained complete remission in pediatric AML patients

doi: 10.1101/2025.07.17.664572

Figure Lengend Snippet: Lymphocyte profiling of the L-TME in pediatric AML patients. (a) Logical gating strategy showing the T cell composition of L-TME in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. The data are displayed cumulatively in the dot plots and individually in the graphs (c and e). The same cumulative and individual representation is depicted in all figures showing flow-cytometry data. (b) I) Cumulative plots displaying CD45 + /CD34 − /CD19 − and CD3 − myeloid cells in AML patients with (top row, n=4 ) and without (bottom row, n=5 ) sustained-CR. II-III) Individual values of samples in (a) and (b) are displayed in a graph; Q value is the result of FDR-corrected multiple t-tests between sustained (red) and non-sustained (black) patients’ samples. Only significant differences are shown. IV) The % of blasts scored in the patients’ diagnosis for all the cohort is displayed in the graph for patients with and without sustained-CR. The blast score was calculated by a combination of microscope and clinical flow-cytometry analyses and provided as clinical report for patients in the Pathology Laboratory of Sidra. (c) I) Phenotype of the CD4 dim cell population: the majority of the cells are IFNγ + (shown in the colored Z-axis) CD4 + and CD8 + doublets, engaging in close networking interactions. A full panel of expression markers for this population is shown in Supplementary Figure 1b. II) Individual values of samples in (b) show a trend of enrichment in CD4 dim doublets (considering lower singlets) in patients without long-term remission.

Article Snippet: Statistical analyses of the flow cytometry data were performed using GraphPad Prism (v. 10.3.1).

Techniques: Flow Cytometry, Biomarker Discovery, Microscopy, Expressing