Fig. 2 In situ vaccination of CpG in combination with anti-OX40 antibody cures established local and distant tumors. In situ vaccination of CpG in combination.

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Fig. 2 In situ vaccination of CpG in combination with anti-OX40 antibody cures established local and distant tumors. In situ vaccination of CpG in combination with anti-OX40 antibody cures established local and distant tumors. (A) Treatment schema. BALB/c mice were implanted subcutaneously with A20 lymphoma cells (5 × 106) on both the right and left sides of the abdomen. When tumors reached between 0.5 and 0.7 cm in the largest diameter (typically on days 4 to 5 after inoculation), αOX40 (4 μg) and CpG (50 μg) were injected into one tumor site every other day for a total of three doses. Tumors sizes were serially measured with a caliper. (B) Tumor growth curves. Left column: Treated tumors (Tr). Right column: Nontreated tumors (NT). Top to bottom: Vehicle, CpG, αOX40, and CpG and αOX40 and survival plots of the treated mice (n = 10 mice per group). ****P < 0.0001, unpaired t test. Shown is one representative experiment out of nine. (C) Effect of CD4/CD8 depletion. Mice were implanted with bilateral tumors, and one tumor was injected with CpG and αOX40 antibody according to the schema in (A). CD4 (0.5 mg)–and CD8 (0.1 mg)–depleting antibodies were injected intraperitoneally on days 6, 8, 12, and 15 (n = 10 mice per group). (D) CD8 T cell immune response. Splenocytes from the indicated groups obtained on day 7 after treatment were cocultured with media, 1 × 106 irradiated 4T1 cells (unrelated control tumor), or A20 cells (homologous tumor) for 24 hours. Intracellular IFN-γ was measured in CD8+ T cells by flow cytometry as a percentage of CD44hi (memory CD8) T cells shown in dot plots and bar graph, summarizing data from three experiments (n = 9 mice per group). ****P < 0.0001, unpaired t test. Idit Sagiv-Barfi et al., Sci Transl Med 2018;10:eaan4488 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works