Alkali injury–induced pathological lymphangiogenesis in the iris facilitates the infiltration of T cells and ocular inflammation
Zheng Liu, Keli Liu, Shunhua Shi, Xun Chen, Xinyu Gu, Weifa Wang, Keli Mao, Rukeye Yibulayi, Wanwen Wu, Lei Zeng, Weibin Zhou, Xiaofeng Lin, Feng Zhang, Bingsheng Lou
Zheng Liu, Keli Liu, Shunhua Shi, Xun Chen, Xinyu Gu, Weifa Wang, Keli Mao, Rukeye Yibulayi, Wanwen Wu, Lei Zeng, Weibin Zhou, Xiaofeng Lin, Feng Zhang, Bingsheng Lou
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Research Article Immunology Ophthalmology

Alkali injury–induced pathological lymphangiogenesis in the iris facilitates the infiltration of T cells and ocular inflammation

Abstract

Inflammatory lymphangiogenesis is intimately linked to immune regulation and tissue homeostasis. However, current evidence has suggested that classic lymphatic vessels are physiologically absent in intraocular structures. Here, we show that neolymphatic vessels were induced in the iris after corneal alkali injury (CAI) in a VEGFR3-dependent manner. Cre-loxP–based lineage tracing revealed that these lymphatic endothelial cells (LECs) originate from existing Prox1+ lymphatic vessels. Notably, the ablation of iridial lymphangiogenesis via conditional deletion of VEGFR3 alleviated the ocular inflammatory response and pathological T cell infiltration. Our findings demonstrate that iridial neolymphatics actively participate in pathological immune responses following injury and suggest intraocular lymphangiogenesis as a valuable therapeutic target for the treatment of ocular inflammation.

Authors

Zheng Liu, Keli Liu, Shunhua Shi, Xun Chen, Xinyu Gu, Weifa Wang, Keli Mao, Rukeye Yibulayi, Wanwen Wu, Lei Zeng, Weibin Zhou, Xiaofeng Lin, Feng Zhang, Bingsheng Lou

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Figure 5

Iridial neolymphatics promote phagocytosis, antigen presentation, and T cell infiltration/activation following CAI.

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Iridial neolymphatics promote phagocytosis, antigen presentation, and T ...
(A and B) GSEA showing expression profiles of phagocytosis/antigen presentation/DC/macrophage/T cell–related immunogenic pathways in the iris tissues following CAI and VEGFR3 depletion. Padj < 0.05 (after Benjamini-Hochberg multiple testing corrections) is considered statistically significant. NES, normalized enrichment score. (C) CD3 and Lyve1 staining of the iris tissue whole mounts from VEGFR3iLECko and VEGFR3fl/fl mice that received CAI at week 6, followed by TAM administered 5 times i.p. between days 13 and 17 and analysis on day 28 after CAI. (D) Quantification of CD3+ T cell counts (per 20× field) in C. Data are shown as mean ± SEM. n = 7 mice per group. Each dot represents 1 mouse. **P < 0.01. Mann-Whitney U test. (E) CD3 and Lyve1 staining of the iris tissue whole mounts from WT mice that received CAI at week 6, followed by VEGFC-156S or PBS intracameral injections administered 3 times between days 16 and 18 and analysis on day 28 after CAI. Note that intracameral injections with PBS alone could alleviate T cell infiltration to the injured iris and that VEGFC-156S administration could promote T cell infiltration following. (F) Quantification of CD3+ T cell counts (per 20× field) in E. Data are shown as mean ± SEM. n = 7 mice per group. Each dot represents 1 mouse. ***P < 0.001. Mann-Whitney U test. (G and H) GSEA showing expression profiles of the indicated antigen presentation/T cell–related immunogenic pathways in the iris tissues following CAI and VEGFR3 depletion. Padj < 0.05 (after Benjamini-Hochberg multiple testing corrections) is considered statistically significant. NES, normalized enrichment score. (I) CD4, CD8, and FOXP3 staining of the iris tissues from VEGFR3iLECko and VEGFR3fl/fl mice that received CAI at week 6, followed by TAM administered 5 times i.p. between days 13 and 17 and analysis on day 28 after CAI. (J) Quantification of CD4+, CD8+, and FOXP3+ T cell counts (per 20× field) in I. Scale bars: 100 μm (C, E, and I). Data are shown as mean ± SEM. n = 6 mice per group. Each dot represents 1 mouse. *P < 0.05, **P < 0.01. Mann-Whitney U test.