Fig. 5: Lysophosphatidic acid rewires CD8 T cell metabolism and modulates reactive oxygen species. | Nature Communications

Fig. 5: Lysophosphatidic acid rewires CD8 T cell metabolism and modulates reactive oxygen species.

From: Lysophosphatidic acid modulates CD8 T cell immunosurveillance and metabolism to impair anti-tumor immunity

Fig. 5

A Mass spectrometry showing global metabolomic data on effector CD8 T cells given media without LPA (RPMI+Glutamine) or treated with 1 µM LPA for 30 min, 2 h, or 4 h prior to sample collection where n = 6 mice per group and shows Euclidean clustering analysis. B Metabolite set enrichment analysis (MSEA) performed on raw data with KEGG analysis to determine enriched metabolic pathways. C Metabolic pathway of γ-glutamyl cycle where blue represents the recycled atoms from γ-L-glutamyl-D-alanine to synthesize glutathione. D–G Relative intracellular abundancies of (D) γ-L-glutamyl-D-alanine and exact p-values are as follows, RPMI+Glutamine vs 30 min LPA p = 0.0313; 30 min LPA vs 2 h LPA p = 0.0347; 30 min LPA vs 4 h LPA p = 0.0290, (E) 5-oxoproline and exact p-values are as follows, 2 h LPA vs 4 h LPA p = 0.0143, (F) L-glutamate and exact p-values are as follows, 2 h LPA vs 4 h LPA p = 0.0262, and (G) glutathione with n = 6 mice per group. H, I Direct measurements of H2O2 in effector CD8 T cells after LPA treatment (H) at 1 µM for 30 min, 2 h, or 4 h or (I) at varying concentrations of LPA after 15 min of LPA treatment with n = 3 mice per group. Data measuring reactive oxygen species are normalized to cells cultured in the absence of LPA. J, K Measurements of lipid peroxidation in effector CD8 T cells after LPA treatment at 1 µM for 30 min, 2 h, or 4 h in (J) OT-I effector CD8 T cells and (K) Lpar5−/− OT-I CD8 T cells with n = 3 mice per group. For (H–K) samples were measured in technical triplicates and error was propagated to biological replicate error where n = 3 mice per group performed in 3 independent experiments. Exact p-values for (H) are as follows, 0 vs 30 min LPA p = 0.0239; 30 min LPA vs 2 h LPA p = 0.0045. Exact p-values for (J) are as follows, RPMI+Glutamine vs 30 min LPA p = 0.0039; 30 min LPA vs 4 h LPA p = 0.0166. Statistics for this entire figure were performed using an ANOVA statistical test with Tukey’s post-hoc analysis was performed where *p < 0.05 and **p < 0.005.

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