Journal: Nature Immunology

Article Title: Cell cycle arrest enhances CD8 + T cell effector function by potentiating glucose metabolism and IL-2 signaling

doi: 10.1038/s41590-025-02407-0

Figure Lengend Snippet: a , Representative proliferation plots of unstimulated mouse CD8 + T cells, or ex vivo-stimulated mouse CD8 + T cells that were either left untreated (non-arrested), arrested in the cell cycle using RO-3306, or released from RO-3306-induced arrest. b , Percentage of viable CD8 + T cells treated or not with HU and RO-3306, measured as Zombie-NIR − cells within the singlet gate ( n =5). Statistical comparisons were performed using RM ANOVA with Sidak’s multiple comparisons; P values are shown on the graphs. c-e , Mouse CD8 + T cells were left unstimulated or ex vivo-stimulated and either non-arrested, cell cycle-arrested with HU or RO-3306, or released from arrest ( n =5). c , Contour plots showing the distribution of cells from each condition. d , Expression levels of the indicated markers overlaid on the opt-SNE map. e , CFSE intensity overlaid on the opt-SNE embedding, where black indicates CFSE low (highly proliferating cells), and copper indicated CFSE high (undivided/first cycle). f , Representative FSC/SSC plots of human and mouse CD8 + T cells that were unstimulated, cell cycle arrested with HU, non-arrested or released from arrest. Percentage of blasted cells (large, activated cells with increased FSC/SSC) is indicated. g , Representative microscopy images of ex vivo-stimulated mouse CD8 + T cells treated transiently with HU or left untreated (3 independent experiments with similar results). Representative FSC/SSC plots of human CD8 + T cells that were unstimulated, cell cycle arrested with RO-3306, topotecan, palbociclib or ribociclib, non-arrested or released from arrest. Percentage of blasted cells is indicated.

Article Snippet: Vaccinated mice were treated intraperitoneally (i.p.) twice daily with 100 mg per kg body weight HU for 4 consecutive days, or daily with 2 mg per kg body weight topotecan (Accord), or daily with 150 mg per kg body weight palbociclib (MedChemExpress).

Techniques: Ex Vivo, Expressing, Microscopy

Journal: Nature Immunology

Article Title: Cell cycle arrest enhances CD8 + T cell effector function by potentiating glucose metabolism and IL-2 signaling

doi: 10.1038/s41590-025-02407-0

Figure Lengend Snippet: a , Representative histograms of CD98 expression on unstimulated human CD8 + T cells, or ex vivo–stimulated human CD8 + T cells that were either left untreated (non-arrested), arrested in the cell cycle using hydroxyurea (HU), or released from HU-induced arrest (released). b , Geometric mean fluorescence intensity (gMFI ± SEM) of CD98 expression; each symbol represents a single healthy donor ( n =7). c , ALDOA expression (gMFI ± SEM) in human CD8 + T cells under the same conditions as in (a) ( n = 5). d , Representative FXcycle versus Edu plots of unstimulated and ex vivo-stimulated human CD8 + T cells (HU arrested, non-arrested and HU released). Lower panels show gMFI of PKM expression in G 0/1 , S and G 2 /M cell cycle phases ( n =5 donors). e , Heatmap showing expression of CD98, G6PD, and PKM in E7 49-57 -specific CD8 + T cells in blood and lymph nodes (LN) on day 7 after vaccination with E7 peptide, in mice treated with palbociclib for 4 consecutive days (days 1-4) ( n =6 mice per group). Color scale represents geometric mean; marker-specific range is indicated for individual markers. f , Percentage (mean ± SEM) of proliferating human CD8 + T cells following ex vivo stimulation with or without HU treatment for 12h or 60h ( n =6 donors). g , Representative histograms of BODIPY-FL-C16 uptake by unstimulated and ex vivo-stimulated human CD8 + T cells (HU arrested, non-arrested, and released after arrest). h , gMFI (± SEM) of BODIPY-FL-C16 uptake ( n =4 donors). i , Heatmap showing relative FDFT1 expression in E7 49-57 -specific CD8 + T cells from blood and lymph nodes (LN) of mice ( n =6) vaccinated with E7 SLP/CpG and treated with HU or left untreated. Statistical comparisons were performed using repeated measures ANOVA with Sidak’s multiple comparisons ( b, c, f, h ); P values are shown on the graphs.

Article Snippet: Vaccinated mice were treated intraperitoneally (i.p.) twice daily with 100 mg per kg body weight HU for 4 consecutive days, or daily with 2 mg per kg body weight topotecan (Accord), or daily with 150 mg per kg body weight palbociclib (MedChemExpress).

Techniques: Expressing, Ex Vivo, Fluorescence, Marker

Journal: Nature Immunology

Article Title: Cell cycle arrest enhances CD8 + T cell effector function by potentiating glucose metabolism and IL-2 signaling

doi: 10.1038/s41590-025-02407-0

Figure Lengend Snippet: a , Representative histograms of γ-H2AX expression in unstimulated human CD8 + T cells and in ex vivo–stimulated human CD8 + T cells that were either left untreated (non-arrested), cell cycle-arrested using hydroxyurea (HU), palbociclib or ribociclib, or released from arrest (released). b , Percentage of γ-H2AX + CD8 + T cells (mean ± SEM) in unstimulated human CD8 + T cells, or following ex vivo stimulation with or without treatment (as described in a) for 24h or 60h. Each symbol represents an individual healthy donor ( n =4). c , Kinetic analysis of pS6 levels in ex vivo-stimulated human CD8 + T cells that were either left untreated (non-arrested), arrested in the cell cycle using HU, or released from HU-induced arrest ( n =5 donors). d , Heatmap of hierarchically clustered z-scored phosphosite intensities related to the MAPK/c-MYC signaling pathway, measured by mass spectrometry in ex vivo-stimulated human CD8 + T cells that were either released from HU-induced arrest or left untreated (non-arrested) ( n =3 donors). Coupled z-scores are based on normalized phosphosite intensities. The number of phospho-groups per site is indicated in parentheses (3P indicates three or more phospho-groups). e , Ex vivo-stimulated mouse CD8 + T cells treated with HU and subsequently released, or left untreated (non-arrested), with or without the addition of the MYC inhibitor 10058-F4 during proliferation. Percentage of proliferating CD8 + T cells (mean ± SEM) is shown. Lines indicate individual mice ( n =4). Statistical comparisons were performed using repeated measures ANOVA with Sidak’s multiple comparisons ( c ) and two-sided paired t-test ( b , e ); P values are shown on the graphs.

Article Snippet: Vaccinated mice were treated intraperitoneally (i.p.) twice daily with 100 mg per kg body weight HU for 4 consecutive days, or daily with 2 mg per kg body weight topotecan (Accord), or daily with 150 mg per kg body weight palbociclib (MedChemExpress).

Techniques: Expressing, Ex Vivo, Phospho-proteomics, Mass Spectrometry