1. Fig. 3. VEGFR-3 signaling increases infiltration of naïve T cells in a CCR7-dependent manner.
VEGFR-3 signaling increases infiltration of naïve T cells in a CCR7-dependent manner. (A to D) B16-OVA and B16-OVA/VC tumor–bearing mice were treated with control (Iso) or VEGFR-3–blocking antibodies (αR3) starting on the day of inoculation, and tumors were characterized on day 9 by flow cytometry (data represent two independent experiments, n = 5 each). (A) Quantification of tumor-infiltrating conventional CD4+ T cells (conv CD4+; FoxP3−) and CD8+ T cells. (B) Activation status of CD4+ (top) and CD8+ (bottom) T cells. Naïve, CD44−CD62L+; effector/effector memory (EM), CD44+CD62L−; central memory (CM), CD44+CD62L+. (C) Ratio of naïve/EM in infiltrating CD4+ and CD8+ T cells. (D) CCL21 concentration as assessed by enzyme-linked immunosorbent assay (ELISA) in the tumor, dLNs, and ndLNs. (E) Representative image of a lymphangiogenic B16/VC tumor section immunostained for CD4+ T cells (red), CCL21 (green), LECs (Lyve-1, white), and DAPI (blue). Scale bar, 100 μm. (F) Quantification of CCR7+ T cell subsets in untreated B16-OVA and B16-OVA/VC tumors after 14 days (n ≥ 4). (G to J) B16-OVA/VC tumor–bearing mice were treated with Iso or anti-CCR7 (αCCR7)–blocking antibodies on days 0, 3, and 6, and (G to I) tumors were characterized on day 9 by flow cytometry or (J) mice were given adoptive transfer of 106 naïve OT-I CD8+ T cells (n ≥ 6). (G) Representative flow cytometry plots of T cell activation status. (H) Quantification of naïve and CM fractions of conventional CD4+ (left) and CD8+ (right) T cells. (I) Ratio of naïve/EM subsets. (J) Quantification of intratumoral OT-I cells as percentage of overall CD8+ T cells on day 10 after inoculation. *P < 0.05, **P < 0.01, ***P < performed with two-tailed Student’s t test or one-way analysis of variance (ANOVA).
Manuel Fankhauser et al., Sci Transl Med 2017;9:eaal4712
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