Extended Data Fig. 8: Pharmacologic activation of the oxidative pentose phosphate pathway results in modest cell death, proliferative inhibition, and altered T cell differentiation. | Nature Immunology

Extended Data Fig. 8: Pharmacologic activation of the oxidative pentose phosphate pathway results in modest cell death, proliferative inhibition, and altered T cell differentiation.

From: Deficiency of metabolic regulator PKM2 activates the pentose phosphate pathway and generates TCF1+ progenitor CD8+ T cells to improve immunotherapy

Extended Data Fig. 8

a-g, OT-I+Thy1.1 + T cells were activated, labeled with CellTrace Violet, treated with either DMSO (red) or 3 µM glucose-6-phosphate dehydrogenase agonist AG1 (blue), and co-cultured with HKP1-ova-GFP until 4 days post-initial stimulation. a, Representative contour plots for Annexin V (Ann V) and CellTrace Violet expression. b-c, Quantification of viability (b) and CellTrace Violet mean fluorescence intensity (MFI) (c). d, Representative contour plots for CD44 and CD62L expression. e-g, Quantification of: CD44 + CD62L+ as percent of Annexin V- Thy1.1+ (e); Annexin V- viable T cells as percent of CD44/CD62L populations in Thy1.1+ cells (f); and CellTrace Violet MFI in viable CD44/CD62L populations in Thy1.1+ cells (g) in T cells from either DMSO control (red) or AG1 (blue) treatment. h-r, Human PBMCs from two different patients, 19-164 and 19-176, were labeled with CellTrace Violet, and T cells isolated and expanded with either DMSO (red) or AG1 (blue) for 3 days. h, Representative contour plots for Annexin V and CellTrace Violet expression in CD45 + CD3 + CD8 + T cells. i-j, Quantification of viability (b) and CellTrace Violet MFI in viable CD8 + T cells (c). k, Representative contour plots for CD45RA and CD62L expression. l–n, Quantification of: CD45RA/CD62L populations as percent of Annexin V- CD45 + CD3 + CD8+ (l); Annexin V- viable T cells as percent of CD45RA/CD62L populations in CD45 + CD3 + CD8+ cells (m); and CellTrace Violet MFI in viable CD45RA/CD62L populations (n). o, Representative contour plots for CD45RO and CD62L expression. p-r, Quantification of: CD45RO/CD62L populations as percent of Annexin V- CD45 + CD3 + CD8+ (p); Annexin V- viable T cells as percent of CD45RO/CD62L populations in CD45 + CD3 + CD8+ cells (q); and CellTrace Violet MFI in viable CD45RO/CD62L populations (r). Numbers: (a-g) n = 6 biological replicates per group. (h-r) n = 2–3 replicates per patient, 2 patients. Statistics: (b,c,e) Unpaired two-tailed t-tests; (f,g,l,m,n,p,q,r), Two-way ANOVA, Šídák multiple comparisons; (i,j) Multiple unpaired two-tailed t-tests, Holm-Šídák multiple comparisons. Data in (b-c,e-g,i-j,l–n,p-r) are presented as mean ± standard deviation. Abbreviations: Ann V, Annexin V; APC, Allophycocyanin; FITC, Fluorescein isothiocyanate; PECy7, Phycoerythrin-Cyanine7; APCCy7, Allophycocyanin-Cyanine7.

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