Fig. 6
BRAFV600E+ melanomas commonly express TNFR2. A Single-cell melanoma suspensions derived from BRAFV600E+ metastatic lesions were evaluated for TNFR1, TNFR2 and CD271 expression by multi-color flow cytometry. Cells were stained for Live/Dead Red (dead cell exclusion), CD45 (leukocyte exclusion), CD90 (fibroblast exclusion), TNFR1, TNFR2 and CD271. A gating strategy and staining examples from two melanoma patients are shown on the left. Summary of TNFR2 and CD271 distribution observed in 16 patients harboring BRAFV600E+ melanoma (shown as percentages) is depicted on the right. B, C Frequencies of SOX10+ TNFR2+ melanoma, as well as relative TNFR2 expression levels on SOX10+ melanoma cells were elevated by immunofluorescence microscopy using biopsies acquired from patients receiving vemurafenib and high dose IFN-α2b therapy. Data were extrapolated from high resolution whole section scans. Vemurafenib was administered orally for three weeks, after which time the patients received high-dose IFN-α2b concurrently with vemurafenib