Journal: Nature medicine
Article Title: Targeted therapy guided by single-cell transcriptomic analysis in drug-induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms: A case report
doi: 10.1038/s41591-019-0733-7
Figure Lengend Snippet: a, scRNAseq tSNE analysis of DiHS/DRESS and HV skin cells, colour-coded based on origin (n=18,218 cells). b, Heatmap of the top-five genes marking clusters of skin cells (x-axis; from ). Keratinocyte (KC); smooth muscle (SM). c, Frequencies of cells in each cluster, color-coded based on origin. d, Nineteen clusters in were grouped into 7 major subsets based on cell types (DiHS/DRESS cells, n=4,676; HVs cells, n=13,542). Numbers of differentially expressed genes (nDEGs) between DiHS/DRESS- and HV cells within each cell type projected onto a tSNE map. DEG: |log fold change| > 0.5, adjusted p-value < 0.05, Wilcoxon rank sum test. e, Immunohistochemical staining for CD3, CD4, and CD8 in DiHS/DRESS lesional skin. Scale bars, 100 μm. Representatives of six serial sections from one sample. f, tSNE plot for the lymphocyte subcluster, colour-coded based on origin (DiHS/DRESS cells, n=589; HV cells, n=1,148). g, tSNE projections of selected genes (n=1,737 cells). h, Frequency of cells from (f) expressing the displayed genes. i, Immunofluorescence staining with anti-CD3 (green) and anti-CCR10 (red) antibodies or rabbit IgG isotype in DiHS/DRESS lesional skin. Dotted lines denote the boundary between the epidermis (Ep) and dermis. Scale bar, 50 μm. j, Immunofluorescence staining with anti-CD3 (green) and anti-JAK3 (red) in DiHS/DRESS lesional skin. Staining in atopic dermatitis and HV skin sections are shown as comparison. Nuclear labeling with DAPI (blue). Scale bar, 50 μm. k, Hematoxylin and eosin staining (top; H&E) and immunohistochemical staining for phosphorylated STAT1 (pSTAT1) in DiHS/DRESS and HV skin. Scale bar, 20 μm. i-k, Representative of 3 independent experiments.
Article Snippet: After blocking for 45 minutes in PBS with 3% skim milk and 5% normal goat or donkey serum (Jackson ImmunoResearch, West Grove, PA, USA), sections were incubated overnight at 4 °C with primary antibodies: mouse anti-human CD3-Alexa Fluro488 (clone UCHT1, Biolegend, #300454, 1:100), rabbit anti-human CCR10 (polyclonal, Thermo Fisher Scientific, #PA1–21617, 1:25), rabbit IgG isotype (Thermo Fisher Scientific, #31235, 1:250), and rabbit anti-human JAK3 (clone 1A1, Bioss Inc, # bsm-54067R, 1:100).
Techniques: Immunohistochemical staining, Staining, Expressing, Immunofluorescence, Labeling