Journal: Cell Reports Medicine
Article Title: High-dimensional profiling of pediatric immune responses to solid organ transplantation
doi: 10.1016/j.xcrm.2023.101147
Figure Lengend Snippet: Experimental approach and sample processing workflow (A) Patient samples were obtained from a biobank constructed as a part of the Clinical Trials in Organ Transplantation-06 (CTOTC-06) study. Patients were classified as “stable” if they had no evidence of rejection for 12 months before and 12 months after sample collection; patients were classified as “rejection” if they developed biopsy-proven rejection 30 days or less after sample collection. (B) PBMCs from donors were thawed, “barcoded” with palladium isotopes, stained with a combination of intracellular and extracellular markers, and analyzed on a mass cytometer. Normalized and debarcoded mass cytometer data were analyzed as described in . (C) For each patient sample, live singlets were identified based on DNA content, event length, and live/dead staining. Twenty-nine subpopulations were identified using previously published combinations of surface and intracellular markers. (D) The events in the terminally differentiated branches of the tree outlined in (C) from all clinical samples were pooled and used to construct a single-cell UMAP (clustering markers CCR7, CD8, CD45RA, CD25, CD3, CD5, CD4, FOXP3, CD56, CD38, GzmB, CD16, CD19, CD20, CD14, TCRγδ, CD11c, CD25, LAG3) to show the phenotypic relationship between the terminally differentiated populations. Cells are colored by their manually gated population. (E) Hierarchically clustered heatmap visualizes the relationships between populations and correlations between marker expression. The median expression of each marker used for gating was then assessed for each of the populations identified in (C).
Article Snippet: CD14 , 156-Gd , HCD14 , Standard Biotools , 3156019B.
Techniques: Construct, Clinical Proteomics, Transplantation Assay, Staining, Cytometry, Marker, Expressing