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. 2022 Mar 21;11(1):2054758. doi: 10.1080/2162402X.2022.2054758

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© 2022 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Combinations of TLR agonists improve the anti-tumor efficacy of a DNA vaccine in a prostate cancer tumor model. C57BL/6 mice were implanted subcutaneously with TRAMP-C1 tumor cells. Mice were immunized intradermally weekly with control vector (pTVG4) or DNA encoding AR ligand-binding domain (pTVG-AR) and delivered alone or co-delivered with TLR1/2, TLR3 and/or TLR9 agonists. (a) shown are the growth curves for each group (n = 6 animals per group. Asterisks demonstrate significant (p < .05) differences as assessed by linear mixed effects model with Geisser-Greenhouse correction. (b) Survival plots using the time to death or when tumors reached 2 cm³ in size, whichever occurred first, are shown. Asterisks indicate p < .05 as assessed by log-rank test. (c) Animals were treated as in panel A, and tumors were obtained at day 64. Tumors were evaluated by flow cytometry for the number of infiltrating CD45⁺ cells, CD8⁺ T-cells among all live cells, and percentage of T_reg cells (CD25⁺Foxp3⁺) among CD45⁺CD3⁺CD4⁺ cells. Asterisk demonstrates significant (p < .05) differences as assessed by ANOVA. (d) Tumor-infiltrating CD8⁺ T-cells were evaluated for 4–1BB and PD-1 expression by flow cytometry. Median Fluorescence Intensity (MFI) for PD-1 was normalized to the lowest value.