Abstract
We aimed to construct a multi-omic single-cell atlas of Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) by examining gene, protein and T-cell receptor (TCR) expression in blister fluid and skin. We performed 5’ scRNA-TCR-cellular indexing of transcriptome and epitopes (CITE)-sequencing on unaffected and affected skin and/or blister fluid from SJS/TEN patients. Following single-cell sequencing, we performed bioinformatic normalization (CellRanger v6.1.2), transcriptome-based clustering (Seurat v4.1.1), and visual and statistical analyses (scCODA, VGAS). The single-cell atlas includes 176,524 cells, which span 18 scRNA-defined immune- and tissue-relevant subtypes, confirmed by scCITE-seq. CD8+ T-cells were significantly enriched in SJS/TEN blister fluid (scCODA p<0.05). Differential gene expression analyses identified shared signatures of cells in SJS-affected skin and blister fluid. Monocytes/macrophages shared an M1-like inflammatory signature (STAT1, HLA, CXCL10), and CD8+ T-cells a signature aligned with cytotoxicity (GNLY, GZMB), interferon-induced signaling (IFITM1, STAT1), and proteasomal processing (PSMB9, PBMB10). Integrated scTCR-seq identified the same expanded clonotypes in both affected skin and blister fluid; expressed on the same T-cell sub-populations absent from or unexpanded in unaffected skin. This SJS/TEN single-cell atlas acts as an unbiased multi-omic tool for patients clustered by drug and reaction which can be used to define specific tissue-relevant signatures of pathogenic antigen-driven oligoclonal T-cells at the site of tissue damage relevant to development biological markers for diagnosis, prognosis and treatment