Journal: Frontiers in Immunology
Article Title: Targeting PD-L1 in solid cancer with myeloid cells expressing a CAR-like immune receptor
doi: 10.3389/fimmu.2024.1380065
Figure Lengend Snippet: CARIR expression in THP-1 macrophages enabled significant cytotoxicity against PD-L1 + target cells. Non-modified- (Ctrl), CARIR-Δz, or CARIR-z-engineered THP-1 effector cells were co-cultured in the presence of 5ng/ml PMA for 3 days with RM-1 hPD-L1 or WT-RM-1 target cells at effector to target ratio of 5:1. Following the co-culture, the cells were stained with APC anti-human CD11b, Brilliant Violet 605 anti-human PD-L1, and Zombie NIR viability dye. The number of the remaining live CD11b - target cells following the co-culture was quantified by flow cytometry with the use of absolute counting beads. (A) Representative contour plots of triplicate experiments show the percentage change for PD-L1 + , PD-L1 int , and PD-L1 - RM-1 hPD-L1 tumor cells following the co-culture. (B) BAR graphs summarize the percentage data shown on (A) . (C) BAR graphs summarize the absolute cell count data shown on (A) . (D) Representative contour plots of triplicate experiments show the percentage change of RM-1 tumor cells (PD-L1 - ) following the co-culture. (E) BAR graph summarizes the percentage data shown on (D) . (F) BAR graph summarizes the absolute cell count data shown on (D) . Tumor cells were gated on non-beads, live, singlets, and CD11b - . Data were presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 by one-way ANOVA.
Article Snippet: The following monoclonal antibodies (mAbs) and their isotype controls were used for flow cytometry: APC human PD-1, APC human PD-L1, Brilliant Violet human PD-L1, APC human CD86, APC human CD11b, PE human CD247 (CD3z), APC mouse CD11b; PE human EGFR from R&D Systems; APC mouse PD-L1 from Tonbo Biosciences.
Techniques: Expressing, Modification, Cell Culture, Co-Culture Assay, Staining, Flow Cytometry, Cell Counting
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