Identification of drug-specific public TCR driving severe cutaneous adverse reactions

Drug hypersensitivity such as severe cutaneous adverse reactions (SCAR), including Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN), could be life-threatening. Here, we enroll SCAR patients to investigate the T cell receptor (TCR) repertoire by next-generation sequencing. A public αβTCR is identified from the cytotoxic T lymphocytes of patients with carbamazepine-SJS/TEN, with its expression showing drug/phenotype-specificity and an bias for HLA-B*15:02. This public αβTCR has binding affinity for carbamazepine and its structural analogs, thereby mediating the immune response. Adoptive transfer of T cell expressing this public αβTCR to HLA-B*15:02 transgenic mice receiving oral administration of carbamazepine induces multi-organ injuries and symptoms mimicking SCAR, including hair loss, erythema, increase of inflammatory lymphocytes in the skin and blood, and liver and kidney dysfunction. Our results not only demonstrate an essential role of TCR in the immune synapse mediating SCAR, but also implicate potential clinical applications and development of therapeutics.


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Policy information about availability of computer code Data collection PCR amplification and next generation sequencing (NGS) for the TCR repertoire: Illumina Miseq flow cytometry: Cytomics FC500 flow cytometer, CXP software (Beckman Coulter) Surface plasmon resonance analysis: Biacore T200 surface plasmon resonance (SPR) biosensor IL-2 production: enzyme-linked immunospot (ELISPOT) assay (Mabtech) and an ImmunoSpot reader (CTL Cellular Technology)

Data analysis
The TRBV, TRBJ, and CDR3 clonotypes were defined according to the ImMunoGeneTics information (IMGT) database (www.imgt.org). Significant differences between the groups were analyzed using an unpaired, two-tailed Student's t test. Graphs were produced using Graphpad Prism (version 7.02). The heat maps were generated using the built-in R heatmap() function in stats package. The Circos plots were generated by VDJtools software (MiLaboratory). The treemaps were generated by the treemap() function version 1.2.0.1, MATLAB Central File Exchange.
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We enrolled a total of 129 subjects, including 73 patients with SCAR (65 with SJS/TEN and 8 with DRESS), 12 tolerant controls, and 44 healthy donors of Han Chinese Data exclusions No data were excluded.

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The identified NGS TCR result was further confirmed by TaqMan qPCR assay as well as by other functional assays. The developed software produces the same NGS results when run with the same starting conditions/parameters. Randomization Does not apply. This work does not describe an experimental study that requires allocation into groups.

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Flow Cytometry
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