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. 2019 Dec 27;9:20057. doi: 10.1038/s41598-019-56370-6

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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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Therapeutic vaccination with inactivated CD47^−/− cells. (a) The study regime: 10 mice per group were implanted subcutaneously with B16F10 cells and treated with PBS, irradiated B16F10, or irradiated CD47^−/− 3BD9 cells 2 days later. Tumors and lymph nodes were collected and immunophenotyping was performed to analyze various lymphocyte populations. (b) Survival of mice after therapeutic vaccination. p = 0.02 by Mantel-Cox test. (c) Tumor growth in mice from the three therapeutic vaccination regimes. **p < 0.01, ***p < 0.001 by unpaired t test. (d) Regulatory T cells (T-regs), proliferating (Ki67⁺) T cells, and NK cells in the tumor microenvironment, and (e) Macrophage (M1- and M2-type) and dendritic cell subsets in the tumor-draining lymph nodes of mice in the PBS, 3BD9, and B16F10 therapeutic vaccinated groups. In all cohorts n = 10. *p < 0.05, **p < 0.01, ***p < 0.001 by unpaired t test performed on GraphPad Prism. Flow cytometric analysis was performed on FlowJo and cell phenotypes are presented as a percentage of the parent cell population.