Journal: Frontiers in Pharmacology

Article Title: BAY 60-6583 Enhances the Antitumor Function of Chimeric Antigen Receptor-Modified T Cells Independent of the Adenosine A2b Receptor

doi: 10.3389/fphar.2021.619800

Figure Lengend Snippet: BAY 60-6583 enhances cytokine secretion of activated anti-CD133 CAR T cells. Anti-CD133 CAR T cells were co-cultured with U251-CD133OE cells, the effector-to-target cell (E:T) ratio was 4:1. Vehicle and eight small molecules targeting adenosine receptors were added at different final concentrations. After 16 h, the medium was collected and secretion of INF-γ (A) and GM-CSF (B) was detected using AlphaLISA kits. Data were normalized to vehicle control and are represented as the mean ± SD of triplicates from a representative experiment of n = 3 experiments.

Article Snippet: After 16 h, the supernatants of the cultures were collected, and cytokines (TNF-α, IFN-γ, and GM-CSF) were detected using AlphaLISA detection kits (TNF-α, AL208C; IFN-γ, AL217C; and GM-CSF, AL216C) from PerkinElmer, Waltham, MA, United States.

Techniques: Cell Culture, Control

Journal: Frontiers in Pharmacology

Article Title: BAY 60-6583 Enhances the Antitumor Function of Chimeric Antigen Receptor-Modified T Cells Independent of the Adenosine A2b Receptor

doi: 10.3389/fphar.2021.619800

Figure Lengend Snippet: BAY 60-6583-mediated CAR T cell enhancement requires T cell activation. Anti-CD133 CAR T cells (1 × 10 6 ) were activated by co-incubation with 2.5 × 10 5 U251-CD133OE cells (A) or CD3/CD28 beads (B) ; unstimulated CAR T cells (C) were used as control. Vehicle and BAY 60-6583 were added at different final concentrations. After 16 h, the medium was collected and the secretion of GM-CSF, INF-γ, and TNF-α was detected using AlphaLISA kits. Data were normalized to vehicle control and are represented as the mean ± SD of triplicates from a representative experiment of n = 4 experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 by 1-way ANOVA; ns, not significant.

Article Snippet: After 16 h, the supernatants of the cultures were collected, and cytokines (TNF-α, IFN-γ, and GM-CSF) were detected using AlphaLISA detection kits (TNF-α, AL208C; IFN-γ, AL217C; and GM-CSF, AL216C) from PerkinElmer, Waltham, MA, United States.

Techniques: Activation Assay, Incubation, Control

Journal: Frontiers in Pharmacology

Article Title: BAY 60-6583 Enhances the Antitumor Function of Chimeric Antigen Receptor-Modified T Cells Independent of the Adenosine A2b Receptor

doi: 10.3389/fphar.2021.619800

Figure Lengend Snippet: BAY 60-6583 specifically improved activities of CD133-CAR T cells. (A–D) Anti-CD133 CAR T cells were co-cultured with U251-CD133OE luc cells or U251 WT luc cells at different E:T ratios. Co-cultures were performed in the presence of vehicle or BAY 60-6583. After 16 h, the medium was collected, and the secretion of INF-γ (A) , TNF-α (B) , and GM-CSF (C) was detected using AlphaLISA kits (results for the E:T ratio of 4:1 are shown). (D) At 48 h after co-culture, cytotoxicity was determined by detecting the bioluminescence signal. Data are represented as the mean ± SD of triplicates from a representative experiment of n = 5 experiments. **** p < 0.0001 by 2-way ANOVA; ns, not significant (E) CFSE-labeled CAR T cells were co-cultured with irradiated U251-CD133OE cells in the presence of BAY 60-6583 or vehicle control. After 120 h, flow cytometry was used to analyze cell proliferation. Results shown are from a representative experiment of n = 4 experiments.

Article Snippet: After 16 h, the supernatants of the cultures were collected, and cytokines (TNF-α, IFN-γ, and GM-CSF) were detected using AlphaLISA detection kits (TNF-α, AL208C; IFN-γ, AL217C; and GM-CSF, AL216C) from PerkinElmer, Waltham, MA, United States.

Techniques: Cell Culture, Co-Culture Assay, Labeling, Irradiation, Control, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: BAY 60-6583 Enhances the Antitumor Function of Chimeric Antigen Receptor-Modified T Cells Independent of the Adenosine A2b Receptor

doi: 10.3389/fphar.2021.619800

Figure Lengend Snippet: The photo-affinity probe enhanced CAR T cell effects similarly to unmodified BAY 60-6583 (A) The structure of the photo-affinity probe (B–D) Anti-HER2 CAR T cells (1 × 10 5 ) were activated by TransAct (B) or 4 × 10 5 MDA-MB-453 luc cells (C, D) . Probe, BAY 60-6583, or vehicle control was added at the beginning. After 16 h, the medium was collected, and the secretion of INF-γ, TNF-α, and GM-CSF was detected using AlphaLISA kits (B, C) ; 48 h later, cytotoxicity was assessed (D) . Data are represented as the mean ± SD from a representative experiment of n = 3 experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 by 1-way ANOVA; ns, not significant.

Article Snippet: After 16 h, the supernatants of the cultures were collected, and cytokines (TNF-α, IFN-γ, and GM-CSF) were detected using AlphaLISA detection kits (TNF-α, AL208C; IFN-γ, AL217C; and GM-CSF, AL216C) from PerkinElmer, Waltham, MA, United States.

Techniques: Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Active Fraction Combination from Liuwei Dihuang Decoction (LW-AFC) Alleviated the LPS-Induced Long-Term Potentiation Impairment and Glial Cells Activation in Hippocampus of Mice by Modulating Immune Responses

doi: 10.1155/2019/3040972

Figure Lengend Snippet: Effect of LW-AFC on cytokine secretion in the (a) hippocampus and (b) plasma of LPS-treated mice. ∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 vs. con and Student's t -test; ### p < 0.001, ## p < 0.01, and # p < 0.05 vs . mod, one-way ANOVA, and Dunnett's test, mean ± SD, n = 5–8. A, IL-1 β ; B, IL-6; C, MCP-1; D, G-CSF; E, IL-10; F, TNF- α ; G, IL-6; H, IFN γ ; I, IL-12; J, IL-10. con, control; mod, model; INDO, indomethacin.

Article Snippet: The secretion of TNF- α in the cell supernatant was determined using an AlphaLISA mouse TNF- α Kit (PerkinElmer, Waltham, MA, USA) according to the manufacturer's protocols using an EnVisionTM 2104 instrument (PerkinElmer, Waltham, MA, USA).

Techniques: Clinical Proteomics, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Active Fraction Combination from Liuwei Dihuang Decoction (LW-AFC) Alleviated the LPS-Induced Long-Term Potentiation Impairment and Glial Cells Activation in Hippocampus of Mice by Modulating Immune Responses

doi: 10.1155/2019/3040972

Figure Lengend Snippet: Effect of compounds from LW-AFC on cell viability and secretion of TNF- α in LPS-stimulated and non-LPS-stimulated BV-2 cells: (a) the cell viability of non-LPS-stimulated BV-2 cells; (b) the cell viability of LPS-stimulated BV-2 cells; (c) secretion of TNF- α in non-LPS-stimulated BV-2 cells; (d) secretion of TNF- α in LPS-stimulated BV-2 cells. ∗∗∗ p < 0.001 and ∗∗ p < 0.01 vs . con and Student's t -test; ### p < 0.001, ## p < 0.01, and # p < 0.05, vs . mod, one-way ANOVA, and Dunnett's test. Mean ± SD n = 3. con, control; mod, model; INDO, indomethacin; DEX, dexamethasone.

Article Snippet: The secretion of TNF- α in the cell supernatant was determined using an AlphaLISA mouse TNF- α Kit (PerkinElmer, Waltham, MA, USA) according to the manufacturer's protocols using an EnVisionTM 2104 instrument (PerkinElmer, Waltham, MA, USA).

Techniques: Control