American Society of Hematology

Figure 2.

$EBV-infected HSCs were detected in CAEBV patients with multiple assays. (A) PrimeFlow data showing EBV-infected cells were identified in BM Lineage–CD34+ HSC cells, from the patients with CAEBV (subjects 1 and 2). (B) Uniform manifold approximation and projection (UMAP) plot showing the two-dimensional (2D) projection of the BM cells derived from the 3 patients with CAEBV (subjects 3-5) based on the single-cell transcriptomics data. According to marker genes (refer to supplemental Figure 4C) identified in each cell cluster (indicated by different colors), cell types were annotated and their names were displayed in their respective locations. (C) UMAP showing a universal EBV infection in the BM hematopoietic system of the patients with CAEBV (subjects 3-5), with the inset demonstrating the infected cells in the HSC compartment. The EBV-infected cells were depicted in red dots. Notably, the red dots (ie, infected cells) were plotted on the top of the gray ones (ie, uninfected cells), so the exact infection percentages (shown in panel D) were not reflected in this plot. (D) Pie charts showing the percentages of EBV-infected cells in each hematopoietic cell population of the patients with CAEBV (subjects 3-5), and the tree-like structure shows the proportion evolution along the differentiation pathways. (E) Significant proportions of EBV-infected cells in HSC, MPP, NK cell populations of each patient with CAEBV (subjects 3-5), compared with none or nonsignificant proportions of the infected cells of the patient with EBV-HLH (subject 6) and healthy controls (HCs; subjects 7 and 8). The error bars indicate the percentage standard deviation (SD) based on bootstrapping. Please note that in some cell populations of individual samples, there were too few cells measured to make a robust estimation on the infection percentage. For example, there were only 11 measured HSCs from subject 8 (HC-8), and therefore the SD was large (9.6%). Asterisks (∗) above the bars indicate that statistically significant fractions of cells were EBV infected, whereas bars without asterisks show that none or nonsignificant fractions of cells were infected. Refer to supplemental Figure 5E-F for other cell populations from BM and cell populations from PB. (F) The EBV copy numbers quantified by RT-qPCR in the CD34+ compartment derived from patients with CAEBV (subjects 3 and 4) and HCs (subjects 7 and 8). B, B cells; Baso, basophils; CMP, common myeloid progenitors; CLP, common lymphoid progenitors; DC, dendritic cells; Ery, erythrocytes; EryP, erythroid progenitors; GMP, granulocyte-monocyte progenitors; MEP, megakaryocyte/erythroid progenitors; Mk, megakaryocytes; Mono, monocytes; Myelo, myelocytes; Neutro, neutrophils; T, T cells.$

EBV-infected HSCs were detected in CAEBV patients with multiple assays. (A) PrimeFlow data showing EBV-infected cells were identified in BM Lineage^–CD34⁺ HSC cells, from the patients with CAEBV (subjects 1 and 2). (B) Uniform manifold approximation and projection (UMAP) plot showing the two-dimensional (2D) projection of the BM cells derived from the 3 patients with CAEBV (subjects 3-5) based on the single-cell transcriptomics data. According to marker genes (refer to supplemental Figure 4C) identified in each cell cluster (indicated by different colors), cell types were annotated and their names were displayed in their respective locations. (C) UMAP showing a universal EBV infection in the BM hematopoietic system of the patients with CAEBV (subjects 3-5), with the inset demonstrating the infected cells in the HSC compartment. The EBV-infected cells were depicted in red dots. Notably, the red dots (ie, infected cells) were plotted on the top of the gray ones (ie, uninfected cells), so the exact infection percentages (shown in panel D) were not reflected in this plot. (D) Pie charts showing the percentages of EBV-infected cells in each hematopoietic cell population of the patients with CAEBV (subjects 3-5), and the tree-like structure shows the proportion evolution along the differentiation pathways. (E) Significant proportions of EBV-infected cells in HSC, MPP, NK cell populations of each patient with CAEBV (subjects 3-5), compared with none or nonsignificant proportions of the infected cells of the patient with EBV-HLH (subject 6) and healthy controls (HCs; subjects 7 and 8). The error bars indicate the percentage standard deviation (SD) based on bootstrapping. Please note that in some cell populations of individual samples, there were too few cells measured to make a robust estimation on the infection percentage. For example, there were only 11 measured HSCs from subject 8 (HC-8), and therefore the SD was large (9.6%). Asterisks (∗) above the bars indicate that statistically significant fractions of cells were EBV infected, whereas bars without asterisks show that none or nonsignificant fractions of cells were infected. Refer to supplemental Figure 5E-F for other cell populations from BM and cell populations from PB. (F) The EBV copy numbers quantified by RT-qPCR in the CD34+ compartment derived from patients with CAEBV (subjects 3 and 4) and HCs (subjects 7 and 8). B, B cells; Baso, basophils; CMP, common myeloid progenitors; CLP, common lymphoid progenitors; DC, dendritic cells; Ery, erythrocytes; EryP, erythroid progenitors; GMP, granulocyte-monocyte progenitors; MEP, megakaryocyte/erythroid progenitors; Mk, megakaryocytes; Mono, monocytes; Myelo, myelocytes; Neutro, neutrophils; T, T cells.

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