角膜上皮来源的血小板反应蛋白-1在干眼疾病中的免疫调节作用。
The immunoregulatory role of corneal epithelium-derived thrombospondin-1 in dry eye disease.
机构信息
Schepens Eye Research Institute, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong, China.
Schepens Eye Research Institute, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA.
出版信息
Ocul Surf. 2018 Oct;16(4):470-477. doi: 10.1016/j.jtos.2018.07.005. Epub 2018 Jul 25.
PURPOSE
In this study, we examine the expression of corneal epithelium-derived thrombospondin-1 (TSP-1) and its immunomodulatory functions in a validated murine model of dry eye disease (DED).
METHODS
DED was induced in female C57BL/6 using a controlled environment chamber (CEC) for 14 days. mRNA and protein expression of TSP-1 by corneal epithelial cells was quantified using real-time PCR and flow cytometry. Corneal epithelial cells from either naïve or DED mice were cultured with bone marrow derived dendritic cells (BMDCs) in the presence of IFNγ for 48 h, and BMDC expression of MHC-II and CD86 was determined using flow cytometry. Next, either recombinant TSP-1 or anti-TSP-1 antibody was added to the co-culture, and BMDC expression of above activation markers was evaluated. Finally, either DED mice were topically treated with either recombinant TSP-1 or human serum albumin (HSA), and maturation of corneal DCs, expression of inflammatory cytokines, and DED severity were investigated.
RESULTS
mRNA expression of TSP-1 by the corneal epithelium was upregulated in DED. Corneal epithelial cells derived from mice with DED demonstrated an enhanced capacity in suppressing BMDC expression of MHC-II and CD86 relative to wild type mice, and this effect was abrogated by TSP-1 blockade and potentiated by recombinant TSP-1. Finally, topical application of recombinant TSP-1 significantly suppressed corneal DC maturation and mRNA expression of pro-inflammatory cytokines, and ameliorated disease severity in mice with DED.
CONCLUSIONS
Our study elucidates the function of epithelium-derived TSP-1 in inhibiting DC maturation and shows its translational potential to limit corneal epitheliopathy in DED.
目的
在这项研究中，我们研究了角膜上皮细胞衍生的血栓素-1（TSP-1）在已验证的干眼症（DED）小鼠模型中的表达及其免疫调节功能。
方法
使用受控环境室（CEC）将雌性 C57BL/6 诱导为 DED 14 天。使用实时 PCR 和流式细胞术定量测量角膜上皮细胞中 TSP-1 的 mRNA 和蛋白表达。从幼稚或 DED 小鼠的角膜上皮细胞中分离出骨髓来源的树突状细胞（BMDC），在 IFNγ存在的情况下培养 48 小时，并使用流式细胞术确定 BMDC 表达 MHC-II 和 CD86。接下来，将重组 TSP-1 或抗 TSP-1 抗体添加到共培养物中，并评估 BMDC 以上激活标志物的表达。最后，DED 小鼠分别用重组 TSP-1 或人血清白蛋白（HSA）局部治疗，并研究角膜 DC 的成熟度、炎症细胞因子的表达和 DED 的严重程度。
结果
DED 时角膜上皮的 TSP-1 mRNA 表达上调。DED 小鼠的角膜上皮细胞表现出增强的抑制 BMDC 表达 MHC-II 和 CD86 的能力，而 TSP-1 阻断和重组 TSP-1 增强了这种作用。最后，重组 TSP-1 的局部应用可显著抑制角膜 DC 的成熟和促炎细胞因子的 mRNA 表达，并改善 DED 小鼠的疾病严重程度。
结论
我们的研究阐明了上皮细胞衍生的 TSP-1 在抑制 DC 成熟中的功能，并显示了其在限制 DED 中角膜上皮病变中的转化潜力。
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