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. 2022 Jun 15;72(2):325–337. doi: 10.1136/gutjnl-2021-325808

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© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

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Interleukin 10 (IL-10) blockade enhances chimeric antigen receptor T (CAR-T) cell migration and tumour apoptosis. (A) Live staining and fluorescent microscopy of colorectal cancer liver metastases (CRLM) tumour slice culture (TSC) treated with untransduced CAR-T with control, untransduced CAR-T with ⍺IL-10, anti-carcinoembryonic antigen (CEA) CAR-T with control, and anti-CEA CAR-T with ⍺IL-10 show CFSE⁺ CAR-T cells (green) within the tumour and near epithelial carcinoma cells (red) when antigen-specific CAR-T are used in combination with αIL-10. Apoptotic cells are shown when activated cleaved Caspases are present and positive for the SR-FLICA reagent (cyan). Tumour slices were imaged after 1 day of treatment. Scale bar=100 µm. (B) Quantification of the carcinoma cells with a CAR-T cell nearby (defined as within 20 µm) with untransduced control CAR-T cells and anti-CEA CAR-T cells treated concurrently with or without αIL-10. Data points represent each high-powered fields (hpf) area imaged, with 3 hpf imaged in each tumour (n=3 separate human tumour samples). Analysis of variance with multiple comparisons. ***p<0.001. (C, D) Quantification of the percentage of total carcinoma cells that are SR-FLICA⁺, and SR-FLICA⁺ carcinoma cells with a CAR-T cell nearby (defined as within 20 µm) after untransduced control CAR-T and anti-CEA CAR-T cells are treated concurrently with or without αIL-10. Data points represent each hpf area imaged, with 3 hpf imaged in each tumour (n=3 separate human tumour samples). Analysis of variance with multiple comparisons. *p<0.05, **p<0.01, ***p<0.001.