Multiomic single-cell sequencing defines tissue-specific responses in Stevens-Johnson syndrome and toxic epidermal necrolysis
Andrew Gibson, Ramesh Ram, Rama Gangula, Yueran Li, Eric Mukherjee, Amy M. Palubinsky, Chelsea N. Campbell, M. C. Thorne, Katherine C. Konvinse, Phuti Choshi, Pooja Deshpande, Sarah Pedretti, Mark W. Fear, Fiona Wood, Richard T. O’Neil, Celestine N. Wanjalla, Spyros A. Kalams, Silvana Gaudieri, Rannakoe Lehloenya, Samuel Bailin, Abha Chopra, Jason A. Trubiano, Jason A. Trubiano, Jonny Peter, S. Mallal, Elizabeth J. Phillips
Abstract
Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) is a rare but life-threatening cutaneous drug reaction mediated by human leukocyte antigen (HLA) class I-restricted CD8+ T cells. For unbiased assessment of cellular immunopathogenesis, here we perform single-cell (sc) transcriptome, surface proteome, and T cell receptor (TCR) sequencing on unaffected skin, affected skin, and blister fluid from 15 SJS/TEN patients. From 109,888 cells, we identify 15 scRNA-defined subsets. Keratinocytes express markers indicating HLA class I-restricted antigen presentation and appear to trigger the proliferation of and killing by cytotoxic CD8+ tissue-resident T cells that express granulysin, granzyme B, perforin, LAG3, CD27, and LINC01871, and signal through the PKM, MIF, TGFβ, and JAK-STAT pathways. In affected tissue, cytotoxic CD8+ T cells express private expanded and unexpanded TCRαβ that are absent or unexpanded in unaffected skin, and mixed populations of macrophages and fibroblasts express pro-inflammatory markers or those favoring repair. This data identifies putative cytotoxic TCRs and therapeutic targets. Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) is a rare but life-threatening severe cutaneous drug reaction mediated by CD8+ T cells. Here the authors characterise the immune response in skin samples at the site of tissue damage from patients affected with SJS/TEN and compare this to healthy skin or blister fluid and find populations of CD8+ T cell clonotypes expressing cytotoxic mediator molecules.