Maemura K, Yanagawa Y, Obata K, Dohi T, Egashira Y, Shibayama Y, Watanabe M
Department of Anatomy, Osaka Medical College, Japan.
Tissue Antigens. 2006 Mar;67(3):198-206. doi: 10.1111/j.1399-0039.2006.00548.x.
Glutamate decarboxylase (GAD), which has two isoforms, GAD65, and GAD67, is responsible for synthesis of the major inhibitory neurotransmitter, gamma-aminobutyric acid. GAD is expressed predominantly in the central nervous system; recent reports suggest that GAD is also expressed in non-neuronal organs including the pancreas. In the pancreatic islets, GAD serves as one of the autoantigens in type I diabetes mellitus. Recent flow cytometric analyses have shown that a variety of self-antigens, including GAD, are ectopically transcribed and expressed in particular cell populations of the thymus, although consensus concerning the cellular phenotype has not been obtained. The aim of this study was to clarify the localization and cellular phenotype of GAD67-expressing cells in the thymus at a cellular level with a novel approach using GAD67-green fluorescent protein (GFP) knock-in mice, in which GFP is expressed specifically in GAD67-positive cells. GFP-positive cells were detected in the thymic medulla and were identified as epithelial cells by immunohistochemistry. Almost all GFP-positive cells were positive for major histocompatibility complex (MHC) class II antigen staining and were positive for both cytokeratin and Ulex Europaeus Agglutinin I, markers of medullary thymic epithelial cells, but were negative for CD11c, Gr-1, and CD45, markers of dendritic cells, macrophages, and B-lymphocytes, respectively.
谷氨酸脱羧酶(GAD)有两种同工型,即GAD65和GAD67,它负责合成主要的抑制性神经递质γ-氨基丁酸。GAD主要在中枢神经系统中表达;最近的报道表明,GAD也在包括胰腺在内的非神经器官中表达。在胰岛中,GAD是I型糖尿病中的自身抗原之一。最近的流式细胞术分析表明,包括GAD在内的多种自身抗原在胸腺的特定细胞群体中异位转录和表达,尽管尚未就细胞表型达成共识。本研究的目的是使用GAD67-绿色荧光蛋白(GFP)基因敲入小鼠,通过一种新方法在细胞水平上阐明胸腺中表达GAD67的细胞的定位和细胞表型,在这种小鼠中GFP特异性地在GAD67阳性细胞中表达。在胸腺髓质中检测到GFP阳性细胞,通过免疫组织化学鉴定为上皮细胞。几乎所有GFP阳性细胞主要组织相容性复合体(MHC)II类抗原染色呈阳性,细胞角蛋白和欧洲荆豆凝集素I均呈阳性,这两种物质是胸腺髓质上皮细胞的标志物,但分别对树突状细胞、巨噬细胞和B淋巴细胞的标志物CD11c、Gr-1和CD45呈阴性。