Human Corneal Endothelial Cells Expressing Programmed Death-Ligand 1 (PD-L1) Suppress PD-1+ T Helper 1 Cells by a Contact-Dependent Mechanism
Human Corneal Endothelial Cells Expressing Programmed Death-Ligand 1 (PD-L1) Suppress PD-1+ T Helper 1 Cells by a Contact-Dependent Mechanism
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DOI：
10.1167/iovs.08-2536
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发表时间：
2009-01-01
影响因子：
4.4
通讯作者：
Yamagami, Satoru
中科院分区：
文献类型：
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作者：
Sugita, Sunao;Usui, Yoshihiko;Yamagami, Satoru
目的。本研究旨在确定人角膜内皮 (HCE) 细胞是否可以在体外调节旁观者 T 细胞的激活。 HCE细胞系是从原代HCE细胞建立的。靶标激活的 T 细胞使用同种异体 T 细胞和 Jurkat T 细胞系。作为额外的靶点，通过有限稀释从眼房水中建立了来自葡萄膜炎患者的 T 细胞克隆。通过 [(3)H]-胸苷掺入、羧基荧光素琥珀酰亚胺酯掺入或 IFN γ 产生来评估 T 细胞活化的增殖情况。通过流式细胞术、RTPCR或免疫组织化学评估经IFNγ处理的角膜内皮细胞和未经处理的细胞上共刺激分子的表达。通过流式细胞术评估靶T细胞上共刺激受体的表达。使用封闭抗体来消除HCE抑制功能。结果。 HCE 细胞通过细胞接触依赖性机制抑制 CD4(+) T 细胞的体外增殖和 IFN γ 产生。 HCE组成型表达共刺激分子程序性死亡配体1（PD-L1）和PD-L2，并且它们的表达被IFNγ增强。 HCE 有效抑制在葡萄膜炎或角膜内皮炎患者建立的各种活化 T 细胞系和克隆中过度表达 PD-1 的 Th1 细胞的增殖。 PD-L1（而非 PD-L2）的中和 mAb 阻断了 HCE 对 Th1 细胞的抑制作用。结论。 HCE 可通过 PD-1/PD-L1 相互作用损害 Th1 浸润 CD4(+) T 细胞的效应功能和激活。这些数据支持这样的假设：角膜内皮可能通过诱导外周免疫耐受来维持眼前房的特权免疫状态。 （投资眼科可见科学。2009 年；50：263-272）DOI：10.1167/iovs.08-2536
PURPOSE. This study was designed to determine whether human corneal endothelial (HCE) cells could regulate the activation of bystander T cells in vitro.METHODS. HCE cell lines were established from primary HCE cells. Target-activated T cells were used allogeneic T cells and Jurkat T-cell lines. As an additional target, T-cell clones from uveitis patients were established from aqueous humor via a limiting dilution. T-cell activation was assessed for proliferation by [(3)H]-thymidine incorporation, carboxyfluorescein succinimidyl ester incorporation, or IFN gamma production. Expression of co-stimulatory molecules on IFN gamma-treated corneal endothelial and non-treated cells was evaluated by flow cytometry, RTPCR, or immunohistochemistry. Expression of co-stimulatory receptors on target T cells was evaluated by flow cytometry. Blocking antibodies was used to abolish the HCE-inhibitory function.RESULTS. HCE cells suppressed both in vitro proliferation and IFN gamma production by CD4(+) T cells via a cell contact-dependent mechanism. HCE constitutively expressed co-stimulatory molecules programmed death-ligand 1 (PD-L1) and PD-L2, and their expression was enhanced by IFN gamma. HCE efficiently inhibited the proliferation of Th1 cells that overexpressed PD-1 among various activated T-cell lines and clones established from patients with uveitis or corneal endotheliitis. A neutralizing mAb for PD-L1, but not PD-L2, blocked the suppressive effect of HCE on Th1 cells.CONCLUSIONS. HCE can impair the effector functions and activation of Th1 infiltrating CD4(+) T cells via the PD-1/PD-L1 interaction. The data support the hypothesis that corneal endothelium may contribute to maintenance of the privileged immune status of the anterior chamber of the eye by inducing peripheral immune tolerance. (Invest Ophthalmol Vis Sci. 2009; 50: 263-272) DOI: 10.1167/iovs.08-2536