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 6

K14-VEGFR3-Ig mice demonstrate impaired antitumor immunity in response to dermal vaccine delivery.

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K14-VEGFR3-Ig mice demonstrate impaired antitumor immunity in response t...
(A) Experimental schematic. WT or K14-VEGFR3-Ig (Tg) mice were vaccinated intradermally (i.d.) in the forelimbs with either 10 μg ovalbumin (OVA) and 10 μg LPS, or 10 μg LPS alone. After 10 days, B16F10.OVA tumor cells (0.5 × 106) were injected i.d. in the back skin and the vaccine was boosted, at the same dose, on day 15. Mice were sacrificed on day 22 (12 days after tumor implantation). (B) Tumor growth was slower in vaccinated vs. control mice for melanomas in WT (left), but not Tg (right) mice. P values determined by regression analysis. *P < 0.05. (C) Representative plots and (D) quantification of circulating antigen-specific CD8+ T lymphocytes as detected by H-2Kb SIINFEKL pentamer staining on day 12. (E) Representative flow cytometry plots showing intracellular cytokine staining for IFN-γ and (F) quantification of IFN-γ+ CD8+ T cells following peptide restimulation of tumor-draining LN cells on day 12. (G) Representative flow cytometry plots and (H) quantification of tumor-infiltrating antigen-specific CD8+ T cells detected by H-2Kb SIINFEKL pentamer staining on day 12 (n = 5, in at least 2 separate experiments). Box and whisker plots show data from min to max. Statistical analysis with 1-way ANOVA. *P < 0.05.