Single-cell gene expression profiles of the less activated CD8⁺ T-cell subpopulations. αβ-TCR⁺ CD8⁺ PBLs were simultaneously stained for CD8, CCR7, CD45RA, CD27, and CD28. CD8^high T cells identified as (A) naive, (B) CD45RA⁻ (left) or CD45RA⁺ (right) CCR7⁻CD27^high, and (C) T_CM were single-cell sorted, and the expression of multiple genes was determined simultaneously in each individual cell. The accuracy of cell sorting was evaluated by the expression of CD3ϵ mRNA, with only cells positive for this molecule being depicted. Each horizontal row depicts the same individual cell, which is numbered. Each vertical row represents a different gene. Results show the profiles from a single representative donor of the 3 we studied. The genes, ccl5, ccl4, tgfb1, tnf, lta, ifng, prf1, gzma, gzmb, faslg, tgfbr1, tgfbr2, ifngr2, and il10ra, code, respectively, for RANTES, MIP-1β, TGF-β1, TNF-α, TNF-β, IFN-γ, perforin, GZMA, GZMB, Fas-L, TGF-βR1, TGF-βR2, IFN-γR2, and IL-10Rα.