Fig. 1

HLA-B*13:01-dependent activation of DDS-specific CTLs in patients with DDS-induced DIHS. A T lymphocyte activation test was performed by IFN-γ ELISpot release assay for CTLs expanded from PBMC of 5 DDS-induced DIHS patients cultured with DDS or DMSO (solvent control). The CTLs-mediated IFN-γ release displayed in a DDS concentration manner. B allogeneic HLA-B*15:02 and HLA-B*13:02 carriers, then in vitro cultured with CTLs from three patients with DDS-induced DIHS upon DDS treatment. C HLA blocking assay were performed, and the IFN-γ releases of DDS-specific CTLs against different cell lines (C1R, HLA-B*13:01 expressing B-LCL or HLA-B*13:02 expressing B-LCL) in the absence or presence of HLA class I or class II antibodies are shown. D DDS-specific CTL induced HLA-B*13:01 expressing B-LCL cell death, but not induced C1R, HLA-B*13:02 expressing B-LCL or HLA-B*15:02 expressing B-LCL, as measured by means of annexin-V labeling for flow cytometry assay. E Cytotoxicity of DDS-specific CTLs against HLA-B*13:01 expressing B-LCL in the presence of DDS as the concentration increases. F DDS-specific CTLs against HLA-B*13:01 expressing B-LCL transfectants and DDS structure/ HLA-B*13:01-related chemicals, including N-Acetyl DDS (NAD), Sulfadiazine (SFD), and Trichloroethylene(TCE). Values are mean and standard error of the mean. G T lymphocyte activation test was performed the IFN-γ release from PBMC -fixing assays cultured with DDS or DMSO. H Activation of CTL cells were analyzed by C1R (HLA-B*13:01)-washing assays cultured with DDS or DMSO respectively. The entire experiment was repeated thrice. *P < 0.05, **P < 0.01, by two-tailed Student t test. B-LCL, B-lymphoblastoid cell line, CTL, cytotoxic T lymphocyte; DIHS, dapsone-induced hypersensitivity syndrome; PBMC, peripheral blood mononuclear cell