Figure 3. T cell costimulation with CD2 prevents development of an exhausted IL7RloPD1hi phenotype.
(A) Schematic of the magnetic bead system providing variable TCR signal duration/costimulation during in vitro culture. (B-F) Scatterplots illustrating IL7R expression by cell division in unstimulated CD8 T cells (B) and following each of three different costimulation cultures (C-F), as indicated. (G-I) Linear plots showing IL7Rhi population resulting from (G) 36h (black line) v 6d (blue line) anti-CD3/28 stimulation, (H) 6d anti-CD2/3/28 (red line) v 6d anti-CD3/28 (blue line) and from 6d anti-CD2/3/28 with (green line) and without (I, red line) Fc-PDL1. (J) Heatmap showing unsupervized hierarchical clustering of murine CD8 T cell gene expression data8 before (naïve, grey), 8 days (effector, green) or 30 days (memory, red) after acute or >30 days (exhausted, blue) after chronic LCMV infection clustered by a CD2 response signature. (K, L) Scatterplots showing GSEA enrichment for genes up (red) and downregulated (blue) by CD2 in (K) memory v exhausted and (L) effector v exhausted CD8 T cells. (M-O) Heatmap showing unsupervized hierarchical clustering of AAV (M, n=58), SLE (N, n=23) and IBD (O, n=58) CD8 T cell expression data using the CD2 response signature. ‘Exhausted’ (blue) and ‘non-exhausted’ (red) subgroups were defined from the major division of the cluster dendrogram. Upper bar indicates comparison with patient subgroups produced using the murine LCMV exhaustion signature (as shown in Fig.2D, G, J). Enrichment by GSEA of CD2 signature in autoimmune subgroups < FDR q 0.1.