Evidence for the targeting by 2-oxo-dehydrogenase enzymes in the T cell response of primary biliary cirrhosis.
Evidence for the targeting by 2-oxo-dehydrogenase enzymes in the T cell response of primary biliary cirrhosis.
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DOI：
10.4049/jimmunol.146.1.89
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发表时间：
1991-01
影响因子：
4.4
通讯作者：
J. V. D. Water;A. Ansari;C. Surh;R. Coppel;Thomas E Roche;H. Bonkovsky;M. Kaplan;M. Gershwin
中科院分区：
文献类型：
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作者：
J. V. D. Water;A. Ansari;C. Surh;R. Coppel;Thomas E Roche;H. Bonkovsky;M. Kaplan;M. Gershwin
原发性胆汁性肝硬化（PBC）是一种慢性自身免疫性肝病，包括门脉系统淋巴浸润，高滴度抗丙酮酸脱氢酶-E2（PDH-E2）和支链酮酸脱氢酶-E2（BCKD-E2）的自身抗体，以及胆道破坏。自身免疫反应的诱导机制、胆管上皮损伤的特异性以及T细胞在PBC中的作用仍不清楚。为了解决这些问题，我们利用了小鼠单克隆抗体，创造了C355.1，并研究了其对正常受试者的一组肝组织以及PBC，进行性硬化性胆管炎和慢性活动性肝炎（CAH）患者的一组肝标本的反应性。C355.1，很像针对PDH-E2的人自身抗体，通过免疫印迹与PDH-E2排他性地反应，结合蛋白质的内部硫辛酰基结构域，并在体外抑制PDH-E2活性。此外，我们还尝试开发克隆的T细胞系，与PDH-E2和/或BCKD-E2反应，使用PBC患者的肝活检，与CAH相比。尽管单克隆C355.1在正常肝脏、胰腺、肺、心脏、甲状腺和肾脏的切片上产生典型的线粒体荧光，但当用于染色PBC患者的胆管时，它产生了明显和强烈的反应性。本文研究的13个PBC肝活检组织中有9个在光学显微镜下含有胆管，所有这些都在1：100的单克隆C355.1培养上清液稀释液中强烈反应。相比之下，尽管正常人、CAH和进行性硬化性胆管炎肝标本的胆管也与C355.1反应，但这种反应性仅为线粒体，仅在1：2稀释时才容易检测到。更重要的是，我们从PBC患者而不是CAH患者中产生了CD 4+、CD 8-、α β TCR+克隆T细胞系，这些细胞系特异性地响应于PDH-E2或BCKD-E2产生IL-2。
Primary biliary cirrhosis (PBC) is a chronic autoimmune liver disease that includes the presence of lymphoid infiltrates in portal tracts, high titer autoantibodies against pyruvate dehydrogenase-E2 (PDH-E2) and branched chain ketoacid dehydrogenase-E2 (BCKD-E2), and biliary tract destruction. The mechanism by which the autoimmune response is induced, the specificity of damage to the biliary epithelium, and the role of T cells in PBC are still unknown. To address these issues, we have taken advantage of a mouse mAb, coined C355.1, and studied its reactivity against a panel of liver tissue from normal subjects as well as a panel of liver specimens from patients with PBC, progressive sclerosing cholangitis, and chronic active hepatitis (CAH). C355.1, much like human autoantibodies to PDH-E2, reacts exclusively by immunoblotting with PDH-E2, binds to the inner lipoyl domain of the protein, and inhibits PDH-E2 activity in vitro. In addition, we have also attempted to develop cloned T cell lines that react with PDH-E2 and/or BCKD-E2 using liver biopsies from patients with PBC, compared with CAH. Although monoclonal C355.1 produced typical mitochondrial fluorescence on sections of normal liver, pancreas, lung, heart, thyroid, and kidney, it produced a distinct and intense reactivity when used to stain the bile ducts of patients with PBC. Nine of 13 PBC liver biopsies studied herein contained bile ducts on light microscopy, all of which reacted intensely at a 1:100 culture supernatant dilution of monoclonal C355.1. In contrast, although bile ducts of liver specimens from normals, CAH, and progressive sclerosing cholangitis also reacted with C355.1, such reactivity was exclusively mitochondrial and readily detectable only at a dilution of 1:2. More importantly, we generated CD4+, CD8-, alpha beta TCR+ cloned T cell lines from patients with PBC, but not from CAH, that produced IL-2 specifically in response to PDH-E2 or BCKD-E2.