T-Cell Suppression by Programmed Cell Death 1 Ligand 1 on Retinal Pigment Epithelium during Inflammatory Conditions
T-Cell Suppression by Programmed Cell Death 1 Ligand 1 on Retinal Pigment Epithelium during Inflammatory Conditions
复制标题
DOI：
10.1167/iovs.08-2846
复制
发表时间：
2009-06-01
影响因子：
4.4
通讯作者：
Mochizuki, Manabu
中科院分区：
文献类型：
--
作者：
Sugita, Sunao;Usui, Yoshihiko;Mochizuki, Manabu
目的。确定视网膜色素上皮 (RPE) 细胞是否可以在炎症条件下抑制体外 T 细胞活化。方法。原代培养的 RPE 细胞是从正常 C57BL/6 小鼠中建立的。目标旁观者 T 细胞是用抗 CD3 抗体从正常脾 T 细胞中建立的。通过检查 [H-3]-胸苷掺入和干扰素 (IFN) γ 或 IL-17 的产生（通过 ELISA 测定）来评估 T 细胞活化的增殖情况。使用寡核苷酸微阵列、RT-PCR、免疫染色和流式细胞术评估 RPE 或重组小鼠 IFN γ 预处理的 RPE 细胞上程序性细胞死亡 1 配体 1 (PD-L1) 的表达。通过流式细胞术评估靶T细胞上程序性细胞死亡1（PD-1）（+）的表达。检测中使用了抗小鼠 PD-L1 或 PD-L2 中和抗体或来自 PD-1 敲除供体的靶 T 细胞。 结果。 IFN γ 预处理的 RPE 极大地抑制了旁观者 T 细胞的激活，尤其是靶 T 细胞（Th1 细胞，但不是 Th17 细胞）通过直接细胞接触产生的 IFN γ 。通过检查细胞表面候选分子，与未处理的对照 RPE 相比，经过 IFN γ 预处理的 RPE 表达的 PD-L1 水平要高得多。尽管原代RPE不表达共刺激分子，但在IFNγ预处理的RPE表面上诱导了该分子的表达。 PD-L1(+) RPE 在 IFN gamma 存在下选择性抑制 PD-1(+) T 细胞活化。在存在抗 PD-L1 中和抗体（但不存在抗 PD-L2）的情况下，经过 IFN γ 预处理的 RPE 未能抑制 T 细胞产生 IFN γ。此外，这些RPE细胞未能抑制来自PD-1无效供体的CD4(+)T细胞产生IFNγ。结论。仅当 RPE 表达负共刺激物时，培养物中的 T 细胞活化才会受到抑制。因此，作者提出，在体外，暴露于 Th1 细胞因子的眼部常驻细胞可以表达该分子，正是这种表达导致了旁观者 Th1 型细胞的抑制。 （投资眼科可见科学。2009 年；50：2862-2870）DOI：10.1167/iovs.08-2846
PURPOSE. To determine whether retinal pigment epithelial (RPE) cells can inhibit in vitro T-cell activation during inflammatory conditions.METHODS. Primary cultured RPE cells were established from normal C57BL/6 mice. Target bystander T cells were established from normal splenic T cells with anti-CD3 antibodies. T-cell activation was assessed for proliferation by both examining [H-3]-thymidine incorporation and the production of interferon (IFN)gamma or IL-17, as determined by ELISA. Expression of programed cell death 1 ligand 1 (PD-L1) on RPE or recombinant mouse IFN gamma-pretreated RPE cells was evaluated using oligonucleotide microarray, RT-PCR, immune staining, and flow cytometry. Expression of programed cell death 1 (PD-1)(+) on target T cells was evaluated by flow cytometry. Anti-mouse PD-L1 or PD-L2 neutralizing antibodies or target T cells from PD-1 knockout donors were used for the assay.RESULTS. IFN gamma-pretreated RPE greatly suppressed activation of bystander T cells, especially the IFN gamma production by the target T cells (Th1 cells, but not Th17 cells) via direct cell contact. By examining cell surface candidate molecules, IFN gamma-pretreated RPE expressed much higher levels of PD-L1 compared with the control nontreated RPE. Although primary RPE did not express the costimulatory molecule, expression of the molecule was induced on the surface of IFN gamma-pretreated RPE. PD-L1(+) RPE in the presence of IFN gamma selectively suppressed PD-1(+) T-cell activation. IFN gamma-pretreated RPE in the presence of anti-PD-L1 neutralizing antibodies, but not anti-PD-L2, failed to suppress T-cell production of IFN gamma. In addition, these RPE cells failed to suppress the production of IFN gamma by CD4(+) T cells from PD-1 null donors.CONCLUSIONS. Suppression of T-cell activation was obtained in cultures only when RPE expressed negative costimulators. Therefore, the authors propose that in vitro, Th1 cytokine-exposed ocular resident cells can express this molecule and it is this expression that causes the suppression of the bystander Th1-type cells. (Invest Ophthalmol Vis Sci. 2009; 50: 2862-2870) DOI: 10.1167/iovs.08-2846