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. 2019 Mar 29;10:1405. doi: 10.1038/s41467-019-09370-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Wnt1 induces cross-tolerance of adoptively transferred transgenic T cells. a Tumor burden (total absolute numbers of Wnt1 overexpressing (Wnt1) vs. control (Empty) OVA-LLC cells) after intrathoracic (i.t.) implantation and adoptive transfer of OVA-specific OTI T cells. b Numbers of OTI T cells per cancer cell (left). IFN-γ expression among adoptively transferred (OTI) and endogenous CD8⁺ T cells (Right). c PD1 expression by intratumoral OTI T cells. d Percentages of TIGIT and TIM-3 expressing intratumoral OTI T cells. e Mean PD1-TIGIT-TIM-3 co-expression depicted in bars (left) and percentages of severely exhausted Tbet^lowEOMES^high cells among PD1^high OTI T cells (Right). f Total absolute numbers of splenic OTI T cells (Up) and proliferation rates assessed by in vivo BrdU incorporation (Bottom). g OVA-LLC cells were co-cultured with intratumoral T cells sorted from Wnt1-overexpressing vs. control OVA-LLC tumors. Relative T cell cytotoxicity: cancer cell death in the presence relative to the absence of T cells. h Equal mixtures of CFSE-low LLC and CFSE-high OVA-LLC cells co-cultured with purified intratumoral OTI T cells from Wnt1-overexpressing vs. control OVA-LLC tumors. Histograms depict percentages of OVA-LLC cells among total LLC cells. i Mice bearing subcutaneous OVA-LLC tumors were adoptively transferred with purified CFSE-labeled OTI T cells from Wnt1-overexpressing vs. control OVA-LLC tumors (Up). Flank tumor growth (bottom left). OTI T cell proliferation measured by CFSE-dilution (Bottom right). j Healthy mice were adoptively transferred with purified OTI T cells from Wnt1-overexpressing vs. control OVA-LLC tumors. k FACS contour plots and graph depict peripheral blood OTI T memory cells 3 months post adoptive transfer. l Flank tumor growth after challenge of the adoptively transferred mice (as in j) with OVA-LLC cells. a–l Cell numbers and profiles were assessed by FACS. Error bars represent mean with SEM. Data are representative or cumulative of at least two independent experiments with 4–8 mice per group. *p < 0.05; **p < 0.01, Mann–Whitney or t-test (K). Source data are provided as a Source Data file