Computational protocol: Armored CAR T cells enhance antitumor efficacy and overcome the tumor microenvironment

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Protocol publication

[…] Flow cytometric analyses were performed using Gallios Flow Cytometer with Kaluza software (Beckman Coulter, Brea, CA, USA). ID8-Muc16ecto and CAR expression were detected using APC-conjugated anti-Muc16 and Armenian hamster 12D11 antibody respectively (Memorial Sloan Kettering Cancer Center monoclonal antibody facility). Mouse cells were stained with rat anti–mouse CD3 (PE/APC. eBioscience 145-2C11/17A2), F4/80 (APC. eBioscience MF48021), CD11b (PE. eBioscience 12-0112-82), PD-L1 (PE. eBioscience 12-5982-82), MHC-II (PE, Thermofisher M5/114.15.2), Granzyme B (PE, Thermofisher 12-8898-82), perforin (PE, Thermofisher 12-9392-82) FAS (PE. eBioscience 12-0951-81) and Ly5.1 (APC. Biolegend; A20). Tumor, mouse T cells or peritoneal cells were pelleted and washed 3 times with FACS buffer (PBS + 2.5% FBS). Cells were resuspended with the appropriate antibody, diluted in FACS buffer and incubated at 4 °C for 30 min in the dark. The cells were subsequently washed 3 times with cold FACS buffer and resuspended in 1X DAPI prior to FACS analysis. Flow sorting of CAR-positive T cells and TAMs was done using a BD Biosciences (San Jose, CA, USA) FACS ARIA IIu Special Order System with BD FACSDIVA software. […]

Pipeline specifications

Software tools Kaluza, BD FACSDiva
Application Flow cytometry
Diseases Neoplasms, Ovarian Diseases, Ovarian Neoplasms, Drug-Related Side Effects and Adverse Reactions