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. 2020 Oct 7;8(2):e000973. doi: 10.1136/jitc-2020-000973

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© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Effects of combination treatment of IL-2 and AKK in ex vivo tumor tissues isolated from patients with CRC. Tumor tissues were dissociated into small pieces, digested and filtrated to generate single-cell suspensions. The cell suspensions were treated with AKK and IL-2 in combination or individually. (A) Schematic illustration of combination treatment of IL-2 and AKK in CRC patient-derived ex vivo tumor tissues. (B) Tumor cells were collected and stained with FITC-conjugated Annexin-V and PI for apoptosis detection by flow cytometry. (C–E) Representative flow cytometry analysis of CD8⁺/CD4⁺ ratio in CD3⁺ T cells (C), activated DCs (D) and cytotoxic effector T cells (E) in tumor-infiltrating lymphocytes isolated from patients with CRC. (F–I) Percentage of apoptosis tumor cells among different groups (F), ratio of CD8⁺/CD4⁺ in CD3⁺ T cells (G), CD80⁺ CD86⁺ in CD11c⁺ cells (H) and IFN-γ⁺ CD8⁺ in CD3⁺ T cells (I). All data are shown as mean±SD (n=3) (**p<0.01). AKK, Akkermansia muciniphila; APC, allophycocyanin; CRC, colorectal cancer; CTL, cytotoxic T lymphocyte; DC, dendritic cell; FITC, fluorescein isothiocyanate; IFN, interferon; IL-2, interleukin-2; PBS, phosphate-buffered saline; PE-PI, phycoerythrin-propidium iodide.