Journal: bioRxiv

Article Title: The natural stilbenoid (–)-hopeaphenol inhibits cellular entry of SARS-CoV-2 USA-WA1/2020, B.1.1.7 and B.1.351 variants

doi: 10.1101/2021.04.29.442010

Figure Lengend Snippet: Identification of stilbenoids as Spike-ACE2 inhibitors by AlphaScreen. A, Demonstration of AlphaScreen-based fluorescence due to interactions of His-tagged Spike RBD (from USA-WA1/2020) and Fc-tagged ACE2 peptides. B, Dose-response curves of stilbenoids and resveratrol on fluorescence inhibition due to disruption of RBD/ACE2 interactions. C, Demonstration of AlphaScreen-based fluorescence due to interactions of His-tagged PD-1 and Fc-tagged PD-L1 peptides. D, Dose-response curves of stilbenoids and control inhibitor BMS-1166 on fluorescence inhibition due to disruption of PD-1/PD-L1 interactions.

Article Snippet: 0.5 nM of human PD-L1-Fc (Sino Biological) was incubated with 5 nM HIS-tagged human PD-1 (Sino Biological) in the presence of 5 μg/mL protein A AlphaScreen acceptor bead and 5 μg/mL nickel chelate donor bead in a total volume of 10 μL of 20 mM HEPES (pH 7.4), 150 mM NaCl, and 0.005% Tween in white, opaque low-volume 384-well plates.

Techniques: Amplified Luminescent Proximity Homogenous Assay, Fluorescence, Inhibition

Journal: Cell Reports Methods

Article Title: Optimization and validation of CAR transduction into human primary NK cells using CRISPR and AAV

doi: 10.1016/j.crmeth.2022.100236

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Article Snippet: 155Gd_PD-1 , Fluidigm , Cat# 3155009B, RRID: AB_2811087.

Techniques: Virus, Recombinant, Electroporation, Antibody Labeling, Labeling, Software

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Increased antitumor activities of glypican-3-specific chimeric antigen receptor-modified T cells by coexpression of a soluble PD1–CH3 fusion protein

doi: 10.1007/s00262-018-2221-1

Figure Lengend Snippet: In vitro behavior of CAR-T cells after repetitive stimulation with SK-HEP-1-GPC3 cells. a CAR expression on the T-cell surface after three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells. b Each data set is pooled from four independent experiments with individual donors (n = 4, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001); c The ratio of CD8+ versus CD4+ T cells after three rounds of antigen-specific stimulation (n = 3, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01); d Proliferation of both CAR-T cells after three rounds of antigen-specific stimulation. The two groups of CAR-T cells were prestained with CFSE before the third stimulation, and then the stained CAR-T cells were cocultured with SK-HEP-1-GPC3 at a 1:1 effector-to-target ratio for 96 h. The intensity of CFSE in each group was measured by flow cytometry. e After three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells, the expression of exhaustion biomarkers on the T-cell surface, including PD-1, TIM-3 and LAG-3, was determined using flow cytometry with the indicated antibodies; f Each data set is pooled from three independent experiments with individual donors (n = 3, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001); g Cytokine release of the engineered T cells after three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells. A total of 1 × 106 engineered T cells were cocultured with 1 × 106 tumor cells for 24 h. The levels of IL-2, IFN-γ and TNF-α in the supernatants were evaluated by ELISA (n = 6, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The level of sPD1 secreted into the culture medium of the transduced T cells was detected using human PD1/PDCD1/CD279 ELISA kits (Sino Biological) according to the manufacturer’s instructions.

Techniques: In Vitro, Expressing, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: Discovery of galectin-8 as an LILRB4 ligand driving M-MDSCs defines a class of antibodies to fight solid tumors

doi: 10.1016/j.xcrm.2023.101374

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Article Snippet: The plate was then washed with PBST (PBS, 0.05% Tween 20) (MA0015, Meilunbio) (T8220, Solarbio) and blocked with 3% BSA (97061-420, VWR) at 37°C for 90 min. After repeated washing, hFc-tagged recombinant proteins, including CD3ε (10977-H02H; Sino Biological), CTLA-4 (CT4-H5255; Acro Biosystems), CD28 (CD8-H525a; Acro Biosystems), CD96 (TAE-H5252; Acro Biosystems), LAG-3 (LA3-H5255; Acro Biosystems), TIM-3 (TM3-H5258; Acro Biosystems), CD40 (CD0-5253; Acro Biosystems), ICOS (ICS-H5258; Acro biosystems), OX40 (OX0-H5255; Acro Biosystems), TIGHT (TIT-H5254; Acro Biosystems), LY86 (10242-H02H; Sino Biological), LILRB4 (16742-H02H; Sino Biological), CD27 (CD7-H5254; Acro biosystems), PD-1 (10377-H02H; Sino Biological), and CD8b (11031-HCCH; Sino Biological), were added into each well.

Techniques: Recombinant, Derivative Assay, Microarray, Enzyme-linked Immunosorbent Assay, Lysis, Blocking Assay, Staining, Transfection, Marker, Flow Cytometry, shRNA, Plasmid Preparation, Negative Control, Silver Staining, Bicinchoninic Acid Protein Assay, Selection, Extraction, Labeling, RNA Expression, Cell Culture, Software