Journal: Oncoimmunology

Article Title: PD-1 checkpoint blockade enhances adoptive immunotherapy by human Vγ2Vδ2 T cells against human prostate cancer

doi: 10.1080/2162402X.2021.1989789

Figure Lengend Snippet: Co-culture with activated Vγ2Vδ2 T cells increases surface expression of PD-L1 on PC-3 cells through a soluble cytokine. PC-3 cells were cultured overnight with or without pamidronate (200 μM) and then washed twice. Treated PC-3 cells were cultured with or without purified Vγ2Vδ2 T cells in either the inner or outer wells in a Transwell plate separated by a 0.4 μm membrane under the indicated conditions for 48 h. PC-3 cells in the outer well were harvested, stained with anti-PD-L1 mAb, and the surface expression of PD-L1 assessed by flow cytometry. Culture supernatants were harvested after 48 h and assayed for IFN-γ levels by ELISA. (a) PD-L1 expression by PC-3 cells cultured under the indicated conditions. Representative of two experiments. Solid histograms show the specific antibody staining while open histograms show control mAb staining. (b) Expression of PD-L1 (MFI) on PC-3 cells in the outer well and supernatant IFN-γ levels after co-culture with Vγ2Vδ2 T cells and PC-3 cells (either untreated or treated with pamidronate) for 48 h

Article Snippet: To determine the effects of interferon on PD-L1 expression on PC-3 cells, PC-3 cells were cultured with either 100 (500 IU), 200 (1,000 IU), or 400 (2,000 IU) ng/ml IFN-γ (ThermoFisher Scientific (Waltham, MA)) or 200 ng/ml (1,000 IU) IFN-α (human hybrid IFN-α A/D, PBL Assay Science (Piscataway, NJ)) for 48 h. To assess BTN3, PD-1, PD-L1, and PD-L2 levels, PC-3 cells were stained with the appropriate mAb and analyzed by flow cytometry.

Techniques: Co-Culture Assay, Expressing, Cell Culture, Purification, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Oncoimmunology

Article Title: PD-1 checkpoint blockade enhances adoptive immunotherapy by human Vγ2Vδ2 T cells against human prostate cancer

doi: 10.1080/2162402X.2021.1989789

Figure Lengend Snippet: IFN-γ and IFN-α increase the surface expression of PD-L1 on PC-3 prostate cancer cells. PC-3 cells were treated with IFN-γ or IFN-α at the indicated concentrations for 48 h. Cells were then stained with various mAbs and surface expression of BTN3, PD-1, PD-L1, and PD-L2 assessed by flow cytometric analysis. (a) Representative histograms of surface expression of various proteins by PC-3 cells cultured under the indicated conditions. Representative of two experiments. Solid histograms show the specific antibody staining while open histograms show control mAb staining. (b) Expression of PD-L1 (MFI) on PC-3 cells after treatment with IFN-γ or IFN-α for 48 h

Article Snippet: To determine the effects of interferon on PD-L1 expression on PC-3 cells, PC-3 cells were cultured with either 100 (500 IU), 200 (1,000 IU), or 400 (2,000 IU) ng/ml IFN-γ (ThermoFisher Scientific (Waltham, MA)) or 200 ng/ml (1,000 IU) IFN-α (human hybrid IFN-α A/D, PBL Assay Science (Piscataway, NJ)) for 48 h. To assess BTN3, PD-1, PD-L1, and PD-L2 levels, PC-3 cells were stained with the appropriate mAb and analyzed by flow cytometry.

Techniques: Expressing, Staining, Cell Culture

Journal: Journal of Oncology

Article Title: Abrine Elicits Liver Carcinoma Immunity and Enhances Antitumor Efficacy of Immune Checkpoint Blockade by Modulating PD-L1 Signaling

doi: 10.1155/2022/7609676

Figure Lengend Snippet: Abrine impairs expression of PD-L1. (a, b) RNA levels of PD-L1 and KAT5 in cancer cells under treatment of abrine were checked by qRT-PCR. (c) Liver cancer cells were stimulated with IFN- γ and treated with or without abrine, and PD-L1 expression was detected by flow cytometry. ∗∗ P < 0.01 vs. control.

Article Snippet: Abrine (20 μ M) [ ] and/or 200 ng/mL IFN- γ (Sigma, USA) was added to stimulate cancer cells for 24 hours.

Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry