Expression of Fas-Fas ligand antigens and apoptotic marker APO2•7 by the human conjunctival epithelium.: Positive correlation with class II HLA DR expression in inflammatory ocular surface disorders
Expression of Fas-Fas ligand antigens and apoptotic marker APO2•7 by the human conjunctival epithelium.: Positive correlation with class II HLA DR expression in inflammatory ocular surface disorders
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DOI：
10.1006/exer.1998.0566
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发表时间：
1998-12-01
影响因子：
3.4
通讯作者：
Baudouin, C
中科院分区：
文献类型：
--
作者：
Brignole, F;De Saint-Jean, M;Baudouin, C
Fas抗原(CD95)是一种在诱导细胞凋亡中起主要作用的膜受体，本文研究了Fas、Fas配体、凋亡标记物APO2.7和已知在炎症条件下表达的膜标记物HLADRⅡ类抗原在结膜上皮细胞中的表达。收集65例患者的印迹细胞学标本：20例正常人，15例戴隐形眼镜者，20例接受慢性局部抗青光眼治疗的患者，10例非特异性慢性结膜炎患者。用抗Fas、Fas配体、APO2.7、HLADR抗原的单抗和阴性同型对照，对细胞进行流式细胞术分析。用标准荧光强度的荧光微球记录阳性细胞的百分比和荧光强度，并用抗PAS刺激抗体孵育人结膜细胞系，以检测PAS在体外诱导的细胞凋亡。Fas在所有标本的大部分结膜细胞中均有表达，但荧光定量结果显示，病变眼的Fas配体和APO2.7的表达明显高于正常眼，Fas配体和APO2.7在结膜细胞中的表达不同，但病变眼的细胞比例明显高于正常眼。在这些眼中，也观察到了强烈的HLADR表达，而正常眼的表达水平最低。PAS、Fas Ligand、APO2.7与HLADR水平呈高度正相关。APO2.7表达和DAPI染色证实，抗Fas抗体在体外可诱导结膜上皮细胞强烈的凋亡。本研究证实结膜上皮细胞正常表达Fas抗原及其配体，角膜上皮细胞和角质形成细胞也是如此。NOW细胞荧光定量分析显示炎症眼的表达明显高于正常眼，并证实了眼表面细胞凋亡与炎症途径之间的强相关性。(C)1998年学术出版社。
Fas antigen (CD95) is a membrane receptor that plays a major role in induction of apoptosis, In surface conjunctival epithelial cells the expressions of Fas, Fas ligand, the apoptotic marker APO2.7 and of HLA DR class II antigen, a membrane marker known to be expressed in inflammatory conditions were investigated. Impression cytology specimens were collected in 65 patients: 20 normal ones, 15 contact lens wearers, 20 receiving chronic topical antiglaucoma treatment and 10 with nonspecific chronic conjunctivitis. Cells were processed for now cytometry, using monoclonal antibodies to Fas, Fas ligand, APO2.7, HLA DR antigens and a negative isotypic control. Percentages of positive cells were recorded and levels of fluorescence quantified using fluorescent beads at standardized fluorescence intensities, In addition, a human conjunctival cell line was incubated with anti-Pas stimulating antibodies in order to test Pas-induced apoptosis in vitro. Fas was found in all specimens in most of the conjunctival cells, but quantitation of levels of fluorescence showed a significantly higher expression in pathologic eyes than in normal ones, Fas ligand and APO2.7 were variably expressed by conjunctival cells, but in a significantly higher percentage of cells in pathological eyes than in normal ones. In these eyes a strong expression of HLA DR was also observed, whereas normal eyes showed lowest levels. Highly significant correlations were found between Pas, Fas ligand, APO2.7 and HLA DR levels. Anti-Fas antibodies in vitro induced strong apoptosis in epithelial cells as confirmed by APO2.7 expression and DAPI staining, This study confirms that conjunctival epithelial cells normally express Fas antigen, and more inconstantly its ligand, as do corneal ones or keratinocytes. Fluorescence quantitation by now cytometry showed much higher expression in inflammatory eyes than in normal ones, and demonstrated a strong correlation between apoptotic and inflammatory pathways in the ocular surface. (C) 1998 Academic Press.