Fig. 4

Identification of DDS-specific TCR in patients with DDS-induced SCAR. A, B analysis of 5′RACE-based TCR sequence results were performed in parallel for UMI corrected data. The expression profiles of TCR repertoire in the blister cell and active PBMC from one DDS-induced SJS patients as well as T cells expanded for 21 days or 35 days of 4 DDS-DIHS patients were analyzed by next-generation sequencing. Heatmaps display the expression values of the TRAV and TRBV genes from each sample, which have been normalized to the mean values of the corresponding gene of the 12 healthy donors’ PBMC, respectively. The definition of TRBV and TRBJ was based on the IMGT database. TRAV12-3, TRBV28, and TRBV20-1 were the most frequent TCR V genes in T cells from patients with DDS-induced SCAR. C, D the pairwise overlap Circos plot shows the overlapping frequency and diversity of TRA and TRB clonotypes among the samples of DDS-induced SCAR, respectively. E, F the frequency of TRA and TRB CDR3 sequence among the samples of DDS-induced SCAR, respectively. The different bar of color represents the frequency of TCR CDR3 in different DDS-induced SCAR patients. G A volcano plot of DEGs that are upregulated or downregulated in DDS-expanded T cells from 5 DDS-induced SCAR patients comparing to non-expansion T cells of 5 healthy donors. The DDS-expanded T cells or non-expansion T cells were collected by FACS using CD3 antibody. The red points and gene names represent the SCAR-related genes as reported previously. P value of volcano plot was derived by Wilcoxon rank-sum test. H single-cell TCR sequencing were performed by 10XGenomics in samples, including DDS-expanded T cells from 1 DDS-induced DIHS patient, non- expanded T cells from 2 fever patients and non- expanded T cells form 3 healthy donors. The frequency of paired TRAV12-3/TRBV28 were shown. The larger size of the circle represents the higher frequency of TCR. Ctrl, control; DEGs, differentially expressed genes; DIHS, drug-induced hypersensitivity syndrome; FACS, Fluorescence activated cell sorter; HD, healthy donors; SJS, Stevens-Johnson syndrome; SCAR, Severe cutaneous adverse reaction; TCR, T cell receptor