程序性死亡分子1及其配体在前房相关免疫偏离鼠虹膜睫状体中的表达

摘要

目的 探讨程序性死亡分子1(PD-1)及其配体在前房相关免疫偏离(ACAID)小鼠虹膜睫状体中的表达及其意义.方法 为实验研究.96只正常BALB/c小鼠随机分配建立ACAID组、阳性对照组、阴性对照组及磷酸盐缓冲液对照组,每组均为24只小鼠.通过观察迟发型超敏反应,评价ACAID的形成与否.采用荧光定量实时聚合酶链反应、免疫组织化学法检测4组小鼠虹膜睫状体中PD-1及其配体1(PD-L1)和配体2(PD-L2)mRNA和蛋白的表达.所有实验结果数据的统计学检验均采用方差分析方法.结果 所有ACAID组小鼠均未出现迟发型超敏反应(DTH),表明ACAID诱导成功.PD-1 mRNA的表达在ACAID组中显著增加,与其他3组比较差异有统计学意义(F=248.109,P＜0.05);而PD-L1 mRNA的表达在阳性对照组中显著增加(F=179.033,P＜0.05).PD-1和PD-L1阳性细胞主要分布于虹膜根部和瞳孔缘区,多呈圆形或类椭圆形,二者的蛋白表达水平分别与它们的mRNA表达水平相一致.4组小鼠虬膜睫状体中均未检测到PD-L2 mRNA和蛋白的表达.结论 ACAID小鼠虹膜睫状体中PD-1表达显著增加,可能参与了ACAID的形成.(中华眼科杂志,2008,44:558-562)%Objective To study the expression and possible implication of programmed death 1 ( PD-1 ) and its ligands in the iris-ciliary body from mice with anterior chamber-associated immune deviation (ACAID). Methods This was an experimental study. Twenty-four BALB/c mice were divided into ACAID group, negative controls, positive controls and phosphate-buffer saline controls. ACAID was evaluated by delayed-type hypersensitivity response. Expressions of PD-1 and its ligands (PD-L1, PD-L2) in the irisciliary body from ACAID mice were determined by real-time polymerase chain reaction and immunohistochemical studies. Results Delayed-type hypersensitivity was not detected in ACAID group,indicating that ACAID was induced successfully. The expression of PD-1 mRNA in ACAID group was higher than that in all other groups (F = 248.109, P < 0.05 ). The positive controls showed a high expression of PD-L1 mRNA ( F = 179.033, P < 0.05). PD-1 and PD-L1 positive ceils with round or oval shapes were found in whole iris-ciliary body, especially in the iris base and pupil margin. Expression of PD-1 and PD-L1 proteins were similar to their expression at mRNA levels. Neither mRNA nor protein of PD-I2 was detected in the iris-ciliary body in all groups. Conclusion An increased expression of PD-I in the iris-ciliary body of ACAID mice indicates that this is involved in the induction of ACAID.