Figure 3.
Single-cell secretion measurements of CTLs from individuals with melanoma and healthy donors. (a) Unsupervised clustering of CD8+ T cells from three healthy donors, presented as a heat map. Each row represents a measurement of 12 secreted cytokines from a single cell, with protein labels provided at the bottom. (b) Data from MART-1–specific TCR–transgenic CTL single-cell experiments, presented as a heat map organized via unsupervised clustering. The fluorescence intensity scale for all four heat maps is the same. (c) Phenotyping data from flow cytometry for MART-1–specific TCR–transgenic CTLs. Each bar shows the percentage of cells positive for the specific surface marker. (d) Fluorospot analysis of the MART-1–specific TCR–transgenic T cell population. The curves on top compare SCBC and Fluorospot measurements on fraction (or number) of cells secreting CCL-2, CCL-5, perforin, IL-1β, IL-6 and TNF-α. Representative Fluorospot images are provided at the bottom. (e) Univariate comparison of antigen-specific TCR–transgenic T cells from a subject with melanoma and healthy donor CD8+ T cell culture controls. The lines of the top plot represent the percentage of cytokine-producing cells from each sample. Blue area shows the range detected from healthy donor samples. The one-dimensional scatter plot on the bottom compares the signal intensities measured from the subject’s MART-1–specific TCR–transgenic T cells and the CD8+ T cell background from one representative healthy donor.