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. 2022 Mar 9;10(3):e003402. doi: 10.1136/jitc-2021-003402

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© Author(s) (or their employer(s)) 2022. 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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Myeloid cells from PIK3CA H1047R tumors are recruited by Ccl2 and inhibit CD8⁺ T cell proliferation. (A) Gene expression in control or PIK3CA H1047R MC38 cells in vitro. (B) Ccl2 transcript abundance after treatment with phospho-inositol 3 kinase (PI3K) inhibitor LY294002 measured by quantitative PCR. Data are mean±SD. (C) Tumor volume over time for PIK3CA H1047R or control tumors with single-guide RNA (sgRNA) for Ccl2 or control guide growing in mice treated with programmed cell death protein 1 (PD-1) blockade. Data are mean±SEM; n=5 animals per group. (D) Proliferation of CD8⁺ splenocytes measured by cell trace violet after coculture for 3 days with bone marrow-derived dendritic cells (BMDCs), anti-CD3, and CD45⁺ cells from control or PIK3CA H1047R tumors. Experimental design (left), representative histograms (center), and quantification (right). Data are mean±SD. (E) Proliferation of CD8⁺ splenocytes measured by cell trace violet after coculture for 3 days with BMDCs, anti-CD3, and F4/80⁺ or Ccr2^hi populations sorted from PIK3CA H1047R tumor. Experimental design (left), representative histograms (center), and quantification (right). Data are mean±SD. *P<0.05; **p<0.01; ***p<0.001; ****p<0.0001.