Fig 2: 2-DG inhibited glycolysis and mTOR pathway in OLP-derived T cells. OLP-derived T cells were activated with 1 µg/ml anti-CD3 mAb and 2 µg/ml anti-CD28 mAb for 72 h, then cultured with 1 mM or 5 mM 2-DG for 24 h. 2-DG was dissolved in DMSO. DMSO at a concentration of 0.1% (v/v) was used as the solvent control. (A) Representative images of immunofluorescent staining for LDHA expression in T cells. The fluorescence intensity of LDHA was decreased in the 2-DG groups. 5 mM 2-DG group showed a weaker fluorescence than 1 mM 2-DG group. Red: LDHA. Blue: nucleus. Bar: 50 µm for ×400. (B) Representative images of Western Blot detection for p-mTOR, mTOR, p-4E-BP1, 4E-BP1, HIF1a and PLD2 expression. Grey-scale analysis revealed that the ratios of p-mTOR/mTOR, p-4E-BP1/4E-BP1, HIF1a/ß-actin and PLD2/ß-actin were decreased in 5 mM 2-DG group, rather than in 1 mM 2-DG group. ***, p < 0.001

Fig 3: Association of the expression levels of NLGN2, CD3 and CD8 in breast cancer. Representative images of immunohistochemical staining showing in situ expression of NLGN2, CD3 and CD8 in breast cancer tissue specimens. Left panels, low expression of NLGN2 in breast tumor tissue, and CD3+ and CD8+ T cells in the same tissue. Right panels, high expression of NLGN2 in breast tumor tissue, and CD3+ and CD8+ T cells in the same tissue. The arrows point to CD3+ or CD8+ lymphocytes. All photos are at 400× original magnification.

Fig 4: PD-L2 in PDAC. a Stratification of PD-L2 expression in PDAC cells (scales bar: 20 µm) and in the stroma. b ?Statistical results of correlation between intratumoral and stromal PD-L2 using the ?2 test. c Expression of CD3, CD8 and FOXP3 in PDAC TILs. d Comparison between intratumoral and stromal densities of CD3+, CD8+ and FOXP3+ T cells in PDAC using paired t tests. e Scatter plots with linear regression for 4 immune markers using Pearson’s correlation. f Relation among densities of CD3+, CD8+, and FOXP3+ T cells based on PD-L2 expression levels using the Mann-Whitney U test. * P < 0.05; **P < 0.01; ***P < 0.001

Fig 5: CD8+ T cells infiltrate the cerebrum of ECM mice and induce ferroptotic damage to primary neurons. a CD8+ T cells infiltrating the cerebrum of ECM mice were co-labeled with CD3 (green) and CD8 (red), and detected by immunofluorescence staining. The nucleus was stained with DAPI (blue). b Morphological changes in primary neurons were observed by microscopy (200 × magnification). c Expression of APP and MAP2 in primary neurons, as measured by western blotting, after co-culture with naïve or activated CD8+ T cells. d Expression of APP and MAP2 in primary neurons, as measured by western blotting after co-culture with activated CD8+ T cells with or without Fer-1. e Relative mitochondrial membrane potential (MMP) levels were calculated based on the optical density value and shown as the fold change relative to the blank value. Data are reported as mean ± standard deviation. Each point represents one sample. *p < 0.05 and ***p < 0.001 vs. control or activated CD8+ T cells. f Expression of TfR1 and GPX4 in primary neurons, measured by western blotting after co-culture with activated CD8+ T cells for 2–24 h. APP amyloid-beta precursor protein, ECM experimental cerebral malaria, GPX4 glutathione peroxidase 4, MAP2 microtubule-associated protein 2, TfR1 transferrin receptor 1