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. 2023 Apr 13;12(1):2201147. doi: 10.1080/2162402X.2023.2201147

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© 2023 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 License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 5. — Neoadjuvant ICB combined with T_reg-depletion induces durable systemic T cell activation and extends metastasis-related survival.

(A) Schematic overview of intervention study. Mice were treated as described in Figure 2A. After mastectomy, treatments were discontinued, and mice were monitored for the development of metastatic disease. (B) Kaplan-Meier curve displaying resection-free survival of mice treated as indicated, until the time they underwent mastectomy at tumor size ~120 mm² (n = 7-10). (C) Representative contour plots depicting CD44 and CD62L expression on CD8+ T cells in blood of mice receiving indicated neoadjuvant treatments, analyzed 1-2 days before mastectomy (pre-mastectomy) and 7 days after mastectomy (post-mastectomy). (D-E) Frequency of indicated markers (% of positive cells) gated on CD8+ T cells (D) and CD4+ T_helper cells (E) in blood of mice post-mastectomy, previously treated as indicated (n = 6-8). (F-G) Frequency of PD-1+ CD44+ CD62L⁻ effector CD8+ (F) and CD4+ (G) T cells in the blood of mice treated as indicated, analyzed by flow cytometry at post-mastectomy (n = 5-7). (H) Kaplan-Meier curve showing metastasis-related survival post mastectomy of mice treated with Ctrl+PBS (n = 10), ICB+PBS (n = 6, 1 censored), Ctrl+DT (n = 12, 2 censored), ICB+DT (n = 11, 4 censored). Censored cases indicate mice that were sacrificed due to metastasis-unrelated causes. (I) Representative image and quantification of immunohistochemical staining for CD8 in metastatic lesions in the lungs of mice previously treated in the neoadjuvant setting as indicated, analyzed at metastasis-related endpoint. Counts represent the average of 20-25 randomly selected 32× fields of views across metastatic lesions (n = 4-7 mice/group; scale bar 100 µm). Data in D-G,I show mean ± SEM. P-values were calculated by Log-rank (Mantel-Cox) test (B,H), Two-way ANOVA with Sidak’s correction (D,E), One-way ANOVA with Tukey’s correction (F,G), Kruskal-Wallis with Dunn’s correction (I). ns, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.