Panel A shows barplots comparing predicted cell type abundances between control and 22q11DS groups. The x-axis lists different cell types, and the y-axis shows the percentage of total thymic cells. Control data is in grey, and 22q11DS data is in purple. Panel B presents barplots of predicted abundances of specific cell types (Fb 2, B cells, Tregs, mTEC(2), and mTEC(3)) in different anatomical regions. The x-axis lists regions, and the y-axis shows the percentage of total cells. Panels C, D, and E display immunofluorescence staining images of Tregs, B cells, and KRT14 positive mTECs in thymic tissue samples from control and 22q11DS groups. Cell nuclei are stained with DAPI (blue), Tregs with anti-FOXP3 antibody (red), B cells with anti-CD19 antibody (orange), and KRT14 positive mTECs with anti-KRT14 (green). Panels F, G, and H show boxplots of mean percentages of Tregs, B cells, and KRT14 positive mTECs in the cortex and medulla regions, with control data in grey and 22q11DS data in purple.
Altered cellular composition of the 22q11DS thymus. (A) Barplots showing predicted cell-type abundance. Cell-type abundance is shown as mean percentages of total thymic cells. Controls (n = 8) are shown in gray, and 22q11DS patients (n = 2) are shown in purple. Differences in cell-type abundance were assessed using two-sided Mann–Whitney U tests. (B) Predicted abundance of Fibroblast type 2 (Fb 2), B cells, regulatory T cells (Tregs), medullary epithelial cells, subtype II (mTEC(II)), and medullary epithelial cells, subtype III (IImTEC(III)) in 22q11DS patients (purple, n = 2) and controls (gray, n = 8) shown as mean percentages of total cells in each annotated anatomical region (* indicates P <0.05). (C–E) CODEX immunofluorescence staining of Tregs (C), B cells (D), and Keratin 14 positive medullary epithelial cells (KRT14+ mTECs) (E) in representative thymic tissue samples from 22q11DS patients and controls. Cell nuclei are stained using DAPI (blue). Tregs are stained using anti-FOXP3 antibody (red), B cells are stained using anti-CD19 antibody (orange), and KRT14+ mTECs are stained using anti-KRT14 (green). (F–H) Boxplot showing mean percentages of Tregs (F), B cells (G), and KRT14+ mTECs (H) in cortex and medulla in the CODEX data. Image processing and segmentation were performed in QuPath, followed by Gaussian mixture rescaling and cell phenotyping in Python using Scimap’s phenotype_cells function. Differences in cell abundance between 22q11DS patients (purple, n = 2) and controls (gray, n = 8 for Tregs, n = 5 for B cells, and n = 5 for KRT14+ mTECs) were assessed using two-sided Mann–Whitney U tests.