Fig. 3 In situ vaccination with CpG and anti-OX40 is therapeutic in a spontaneous tumor model. In situ vaccination with CpG and anti-OX40 is therapeutic.

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Fig. 3 In situ vaccination with CpG and anti-OX40 is therapeutic in a spontaneous tumor model. In situ vaccination with CpG and anti-OX40 is therapeutic in a spontaneous tumor model. (A) MMTV-PyMT transgenic female mice were injected into the first arising tumor (black arrow) with either vehicle (top) or with CpG and αOX40 (bottom); pictures were taken on day 80. (B) CpG and αOX40 decrease the tumor size of a nontreated contralateral tumor. Growth curves represent the volume of a contralateral (untreated) tumor in mice that had two palpable tumors at the beginning of treatment. Mice treated by in situ vaccination (red; n = 6) or vehicle (black; n = 6). ***P = 0.0008, unpaired t test. (C) CpG and αOX40 decrease the total tumor load. Growth curves represent the sum of the volume of 10 tumors from the different fat pads of each mouse, measured with calipers (n = 10 mice per group), and the window of treatment is indicated by the gray bar. ****P < 0.0001, unpaired t test. (D) Time-matched quantification of the number of tumor-positive mammary fat pads. **P = 0.011, unpaired t test (n = 9 mice per group). (E) Mice were sacrificed at the age of 80 days, and lungs were excised and analyzed ex vivo for the number of metastases (mets). ****P < 0.0001, unpaired t test (n = 10 mice from vehicle-treated group; n = 9 mice with CpG and αOX40). (F) Survival plots of the treated mice. ****P < 0.0001. Data are means ± SEM (n = 10 mice per group). (G) CD8 T cell immune response. Splenocytes from the indicated groups obtained on days 7 to 15 after treatment were cocultured for 24 hours with either media or 1 × 106 irradiated tumor cells taken from an independent contralateral site on the body. Intracellular IFN-γ was measured in CD8+ T cells by flow cytometry as shown in dot plots and bar graph, summarizing data as a percentage of CD44hi (memory CD8) T cells (n = 3 mice per group). Idit Sagiv-Barfi et al., Sci Transl Med 2018;10:eaan4488 Published by AAAS