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[Preprint]. 2024 Feb 18:2023.11.26.568771. [Version 2] doi: 10.1101/2023.11.26.568771

Figure 1. Unbiased multi-focal single-cell sequencing identifies expanded pathogenic CD8+ Tconv in affected skin and blister fluid during Stevens-Johnson syndrome and toxic epidermal necrolysis.

Figure 1.

(A) (i) scRNA-defined uniform manifold approximation and projection (UMAP) of 119,784 live cells from normal skin, burn blister fluid, and unaffected skin, affected skin and/or blister fluid from 17 patients with acute SJS/TEN. (ii) Expression distribution for key cell types. The majority UMAP location of each subset is labeled with the percentage of each subset within that location indicated. Figure created using VGAS. (B) Integrated scCITE-seq expression of subset-defining cell surface proteins for T-cells (CD3, CD4, CD8), NK cells (CD56), DC (CD1c), monocytes and macrophages (CD14, CD11c), and B cells (CD19). (i) Heatmap expression (high to low, red to blue) for each surface protein is shown on a representative UMAP of 17,629 cells obtained within a single 10x run, and (ii) average box plot expression is shown between scRNA-defined subsets. Figure created using VGAS. (C) Individual scRNA-defined UMAPs for tissue-relevant controls from unrelated donors (burn blister fluid, normal skin) and skin and blister fluid from patients with SJS/TEN. The average expression is shown for patients with multiple samples from the same time point, indicated by an asterisk. Figure created using VGAS. (D) scRNA-defined cell representation of (i) all subsets in individual samples and (ii) rare, unconventional, and innate-like lymphoid cell (ILC) populations as a proportion of total CD8 T-cells, CD4 T-cells, or NK cells in each sample. (E) (i) Seurat-defined Tconv clusters 1–8 with shared naming according to differential gene expression signatures and (ii) proportional representation in control and disease sample phenotypes. (iii) Pathway analysis for the significant differentially expressed gene signature of CD8 Tconv cluster 3 using Enrichr and gene ontology (GO) cellular and biological terms ranked by p-value. MSC, Mesenchymal stromal cell; HSC, Hematopoietic stem cell; NK, Natural killer; DC, Dendritic cell; Tconv, T conventional cell; unconv., unconventional; ILC, innate-like lymphoid cell; Unaff, Unaffected; Aff, Affected.