Fig 1: A molecular subtyping scheme for CTCL based on the transcriptomes of malignant T cells.a Heatmap of unsupervised hierarchical clustering showing the average expression of the DEGs between malignant T cells and their respective reactive CD4+ or CD8+ T cells in each sample from 13 patients in the 10× Genomics dataset (log2 fold change >1.5, p < 0.05). The bars above the heatmap show the tumor type, LCT information and disease stage of each patient. LCT, large cell transformation. b Immunofluorescence staining of CD45RO (green) and CD27 (red) on tumor samples from the TCyEM and TCM groups. DAPI (blue) was used to visualize cell nuclei. Scale bar = 50 µm. Results are representative of three different samples. c Immunofluorescence staining demonstrates that CD4+ tumorous cells (red) express several cytotoxic markers (red). DAPI (blue) was used to visualize cell nuclei. Scale bar = 50 µm. Results are representative of three different samples. d Venn diagrams illustrate the number of overlapping DEGs (log2 fold change >0.25, p < 0.05) of representative TCyEM and TCM patients. The gene signatures are listed. Genes previously reported to be upregulated in CTCL are highlighted in red. e 2D density plots show the cytotoxicity and exhaustion score of malignant T cells in the two groups from the 10× Genomics dataset. f Scatterplots show the T cell activation and proliferation states of all T cells in the two groups from the 10× Genomics dataset. Blue dots represent reactive T cells. Red dots represent malignant T cells. g PFS analysis of the 49 tumor-stage MF patients. Patients were stratified into low and high expression groups according to median values of scores corresponding to the gene signatures of the TCyEM (left) and TCM (right) groups identified in Fig. 4d. P values were calculated using the log-rank test. h Pseudotime trajectory analysis of all CD4+ T cells in CD4+ CTCL patients from the 10× Genomics dataset inferred by Monocle 2, with cells colored by cell types, pseudo-time, molecular subtypes and TCR clonotypes in sequence. Naive T cells were selected as the start cells. Source data for (d) are provided in the Source Data file.
Fig 2: LTA+ B cell subpopulations in human melanoma. Identification of a LTA+ CD19+ CD20+ CD27- CD38- memory-like B cell (white arrow), a LTA+ CD19+ CD20- CD27+ CD38- activated B cell (yellow arrow) and a LTA+ CD19+ CD20- CD27- CD38+ antibody secreting cell (blue arrow). Serial images of the same cells for the different cell markers and the cytokines LTA and IL-10 in the two upper rows; composite images for marker combinations are given in the two lower rows, together with respective LTA staining and negative staining for IL-10. DAPI nuclear staining in the lower right. Arrows depict the same cell, being representative of the respective B cell subpopulation. Scale bar represents 50 µm.
Fig 3: Melanoma TAB are enriched for plasmablast- and plasma cell-like phenotypes. a Marker combinations used to identify TAB subpopulations by seven-color multiplex immunostaining. b Relative frequency of different TAB phenotypes in whole tissue sections of 41 metastatic melanomas c, d Multiplex immunostaining identifies CD19+CD20-CD38+CD138-CD27+ plasmablast-like (c) TAB and CD19+CD20-CD38+CD138+CD27+ plasma cell-like (d) TAB. Here, serial images display the same cells from a stromal area at the invasive tumor margin. A full composite image together with DAPI nuclear staining (bottom right) and images for each of the individual markers and different combinations from the composite image are shown. Arrow depicts one of several plasmablast-like (c) and plasma cell-like (d) TAB. Scale bars represent 20 µm
Fig 4: T cell-B cell conjugates in IgG4-RD end organs(A) Immunofluorescence staining of CD19 (red), CD27 (cyan), IgD (yellow), CD20 (purple), CD4 (green), CD8 (orange), and DAPI (blue) in a submandibular gland (SMG) from a patient with IgG4-RD.(B) Absolute numbers and percentages of IgD-CD27- DN B cells and IgD-CD27+ B cells in IgG4-RD (n = 7). This combines data for which a representative image is shown in Figure S1 and from (A).(C) Absolute numbers of DN B cells (blue), IgD-CD27+ B cells (gold), and plasmablasts (gray) in IgG4-RD (n = 4).(D) Immunofluorescence staining of CD19 (red), CD27 (cyan), IgD (yellow), CD20 (purple), CD4 (green), CD8 (orange), and DAPI (blue) and visualization of physically interacting T and B cells in an SMG from a patient with IgG4-RD using the StrataQuest cell-to-cell contact application. Nuclei circled in red depict CD19+ B cell in physical contact with CD4+ T cells. T cells and B cells formed close and extensive intercellular plasma membrane contacts.(E) Absolute numbers of DN B cells (blue), SWM B cells (yellow), and plasmablasts (purple) in physical contact with CD4+ T cells in patients with IgG4-RD (n = 4).(F) Absolute numbers of DN B cells, SWM B cells, and plasmablasts conjugated with CD4+ T cells (blue) and CD8+ T cells (yellow) in patients with IgG4-RD (n = 4).
Fig 5: Representative IHC staining of CD3, CD8, and CD27 in the same place at the same time, namely in the core of the tumor (CT) and in the tumor’s invasive margin (IM) in colon tumor tissue (100×).
Supplier Page from Abcam for Anti-CD27 antibody [EPR8569]