Lymphatic vessels regulate immune microenvironments in human and murine melanoma
Amanda W. Lund, … , Helge Wiig, Melody A. Swartz
Amanda W. Lund, … , Helge Wiig, Melody A. Swartz
Published August 15, 2016
Citation Information: J Clin Invest. 2016;126(9):3389-3402. https://doi.org/10.1172/JCI79434.
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Research Article Angiogenesis

Lymphatic vessels regulate immune microenvironments in human and murine melanoma

Abstract

Lymphatic remodeling in tumor microenvironments correlates with progression and metastasis, and local lymphatic vessels play complex and poorly understood roles in tumor immunity. Tumor lymphangiogenesis is associated with increased immune suppression, yet lymphatic vessels are required for fluid drainage and immune cell trafficking to lymph nodes, where adaptive immune responses are mounted. Here, we examined the contribution of lymphatic drainage to tumor inflammation and immunity using a mouse model that lacks dermal lymphatic vessels (K14-VEGFR3-Ig mice). Melanomas implanted in these mice grew robustly, but exhibited drastically reduced cytokine expression and leukocyte infiltration compared with those implanted in control animals. In the absence of local immune suppression, transferred cytotoxic T cells more effectively controlled tumors in K14-VEGFR3-Ig mice than in control mice. Furthermore, gene expression analysis of human melanoma samples revealed that patient immune parameters are markedly stratified by levels of lymphatic markers. This work suggests that the establishment of tumor-associated inflammation and immunity critically depends on lymphatic vessel remodeling and drainage. Moreover, these results have implications for immunotherapies, the efficacies of which are regulated by the tumor immune microenvironment.

Authors

Amanda W. Lund, Marek Wagner, Manuel Fankhauser, Eli S. Steinskog, Maria A. Broggi, Stefani Spranger, Thomas F. Gajewski, Kari Alitalo, Hans P. Eikesdal, Helge Wiig, Melody A. Swartz

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Figure 2

Impaired tumor drainage and DC trafficking to local lymph nodes in K14-VEGFR3-Ig mice.

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Impaired tumor drainage and DC trafficking to local lymph nodes in K14-V...
B16F10 tumors were implanted intradermally into WT or K14-VEGFR3-Ig transgenic (Tg) mice and excised at day 9. (A) Peritumoral area from WT or Tg mice stained for lymphatic endothelial cells (LYVE1; n = 3; n.d., not detected) (arrowhead). Scale bar: 50 μm. (B) Lymphatic vessel density quantified as LYVE1+ structures per 0.04 mm2. Data were compared using Student’s unpaired t test. **P < 0.01 (C) Intratumoral area stained for blood endothelial cells (CD31, n = 3) (arrowhead). Scale bar: 50 μm. (D) Blood vessel density quantification as CD31+ structures per 0.04 mm2. (E) Tumor growth profiles in WT and Tg mice over 13 days. (F) Fluid drainage from the tumor to the draining lymph node (dLN, brachial) assessed 30 minutes after intratumoral injection of 70-kDa FITC-dextran and plotted as fluorescence intensity in arbitrary units (A.U.) per LN normalized to WT (n = 7). (G) DC trafficking from the tumor to the dLN, determined by the quantity of CD11c+MHCII+FITC+ cells in the dLN 24 hours after intratumoral injection of 0.5-μm FITC-labeled latex beads and representative flow cytometry dot plots (n = 5). (H) Quantification of bead+ DCs in the LN. (I) Comparison of relative B cell (B220+) and T cell (CD3ε+) populations in the dLNs to nondraining lymph nodes (ndLN) (n ≥ 4). Data are represented as the mean ± SEM. Statistical analysis with Mann-Whitney U test. *P < 0.05, **P < 0.01.