Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The glucocorticoids prednisone and dexamethasone differentially modulate T cell function in response to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade

doi: 10.1007/s00262-020-02555-2

Figure Lengend Snippet: Production of IL-2, IFN-γ and TNF-α by PBMCs stimulated and treated with indicated concentrations of prednisone (pred), anti-PD-1 (pembrolizumab (pem)), anti-CTLA-4 (ipilimumab (ipi)) alone, or in combination. a Plots showing fold concentration of IL-2, b IFN-γ and c TNF-α in cell culture supernatants from PBMCs stimulated by SEB (100 ng/ml) plus the indicated treatments compared to SEB stimulated only control cells. PBMCs were stimulated for 72 h. Data from four different experiments. Error bars represent mean ± SEM. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test. Unstim, unstimulated cells

Article Snippet: T cell stimulation and Phospho-SHP-2 (Y542) ELISA PBMCs were stimulated with anti-CD3 (3ug/ml) and anti-CD28 (1 μg/ml) at a density of 5 × 10 6 cells/ml in a 24-well plate, with or without anti-PD-1 (10 μg/ml), prednisone (1 μg/ml), dexamethasone (0.5 μg/ml) or a combination for 3 h. Cells were harvested and lysed using Cell Extraction Buffer (Thermo Scientific) supplemented with protease inhibitors (cOmplete Protease Inhibitor Cocktail, Roche) and 1 mM PMSF (Sigma).

Techniques: Concentration Assay, Cell Culture, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The glucocorticoids prednisone and dexamethasone differentially modulate T cell function in response to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade

doi: 10.1007/s00262-020-02555-2

Figure Lengend Snippet: Prednisone treatment does not affect expression of PD-1, CTLA-4, TIM-3 and LAG-3 by SEB stimulated CD4+ and CD8+ T cells. a Cumulative data showing expression of PD-1 on CD4+ T cells, b CTLA-4 on CD4+ T cells, c Tim-3 on CD4+ T cells and d LAG-3 on CD4+ T cells. e Cumulative data showing expression of PD-1 on CD8+ T cells, f CTLA-4 on CD8+ T cells, g Tim-3 on CD8+ T cells and h LAG-3 on CD8+ T cells in response to SEB stimulation (100 ng/ml) plus the indicated treatments compared to stimulated-only control cells. PBMCs were stimulated for 72 h. Data from four independent experiments shown. Error bars represent mean ± SEM. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test. Unstim, unstimulated cells, prednisone (pred), anti-PD-1 (pembrolizumab (pem)), a-CTLA-4 (ipilimumab (ipi))

Article Snippet: T cell stimulation and Phospho-SHP-2 (Y542) ELISA PBMCs were stimulated with anti-CD3 (3ug/ml) and anti-CD28 (1 μg/ml) at a density of 5 × 10 6 cells/ml in a 24-well plate, with or without anti-PD-1 (10 μg/ml), prednisone (1 μg/ml), dexamethasone (0.5 μg/ml) or a combination for 3 h. Cells were harvested and lysed using Cell Extraction Buffer (Thermo Scientific) supplemented with protease inhibitors (cOmplete Protease Inhibitor Cocktail, Roche) and 1 mM PMSF (Sigma).

Techniques: Expressing, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The glucocorticoids prednisone and dexamethasone differentially modulate T cell function in response to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade

doi: 10.1007/s00262-020-02555-2

Figure Lengend Snippet: T cell proliferation in response to prednisone treatment and cytokines (IL-2, IFN-γ and TNF-α) production in response to dexamethasone. a CFSE dilution by CD4+ and b CD8+ T cells in response to PBMC stimulation by anti-CD3 (1 μg/ml) plus indicated treatments for 96 h. Data from four independent experiments shown. Error bars represent mean ± SD. P values obtained by Mann–Whitney nonparametric testing for CFSE analysis. (c) Cumulative data showing fold concentration of IL-2, (d) IFN-γ and (e) TNF-α in cell culture supernatants from PBMCs stimulated with SEB (100 ng/ml) plus the indicated treatments compared to SEB stimulated only control cells. PBMCs were stimulated for 72 h. Data from four independent experiments shown. Error bars represent mean ± SEM. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test. Unstim, unstimulated cells, dexamethasone (dexa), anti-PD-1 (pembrolizumab (pem)), a-CTLA-4 (ipilimumab (ipi))

Article Snippet: T cell stimulation and Phospho-SHP-2 (Y542) ELISA PBMCs were stimulated with anti-CD3 (3ug/ml) and anti-CD28 (1 μg/ml) at a density of 5 × 10 6 cells/ml in a 24-well plate, with or without anti-PD-1 (10 μg/ml), prednisone (1 μg/ml), dexamethasone (0.5 μg/ml) or a combination for 3 h. Cells were harvested and lysed using Cell Extraction Buffer (Thermo Scientific) supplemented with protease inhibitors (cOmplete Protease Inhibitor Cocktail, Roche) and 1 mM PMSF (Sigma).

Techniques: MANN-WHITNEY, Concentration Assay, Cell Culture, Control