Transgenic mice expressing osteopontin in hepatocytes as a model of autoimmune hepatitis
Transgenic mice expressing osteopontin in hepatocytes as a model of autoimmune hepatitis
复制标题
DOI：
10.1016/j.bbrc.2004.02.180
复制
发表时间：
2004-04-23
影响因子：
3.1
通讯作者：
Fujiwara, K
中科院分区：
文献类型：
--
作者：
Mochida, S;Yoshimoto, T;Fujiwara, K
骨桥蛋白是Th 1免疫应答的关键因子，在受损肝脏中活化的星状细胞和巨噬细胞中表达。为了阐明骨桥蛋白在肝脏炎症变化中的作用，我们试图建立在肝细胞中表达骨桥蛋白的转基因小鼠。从刀豆蛋白A刺激的C57 BL/6小鼠脾细胞中克隆了小鼠骨桥蛋白cDNA，将其构建到含有血清淀粉样蛋白P组分启动子的载体中。将该构建体显微注射到C57 BL/6小鼠的受精卵中，获得了4个品系的转基因小鼠。Western blotting和免疫组化显示，骨桥蛋白在肝细胞中表达，而不是在非实质细胞，在转基因小鼠。小鼠肝脏和血浆中的平均骨桥蛋白浓度分别是阴性同窝小鼠的13倍和2.6倍。50%的转基因小鼠血浆抗核抗体阳性。在转基因小鼠晚于12周龄，单核细胞浸润在肝脏中的开发，这些细胞是阳性的CD 8和HLA-DR。血浆ALT活性增加，肝小叶局灶性坏死的转基因小鼠晚于24周龄。在肝细胞中表达骨桥蛋白的转基因小鼠可用作自身免疫性肝炎的模型。(C)2004年爱思唯尔公司All rights reserved.
Osteopontin, a crucial factor for Th1 immune response, is expressed in stellate cells and macrophages activated in injured liver. To clarify the role of osteopontin in inflammatory changes in the liver, we attempted to establish transgenic mice expressing osteopontin in hepatocytes. Mouse osteopontin cDNA, cloned from concanavalin-A-stimulated spleen cells in C57BL/6 mice, was constructed into the vector containing serum amyloid-P component promoter. This construction was microinjected into fertilized eggs of C57BL/6 mice, and 4 lines of the transgenic mice were obtained. Western blotting and immunohistochemistry revealed that osteopontin was expressed in hepatocytes, but not in non-parenchymal cells, in the transgenic mice. The mean osteopontin concentrations in the liver and plasma in the mice were 13 and 2.6 times higher than those in negative littermates. Antinuclear antibody was positive in the plasma in 50% of the transgenic mice. In the transgenic mice later than 12 weeks of age, mononuclear cell infiltration in the liver developed, and these cells were positive for CD8 and HLA-DR. Plasma ALT activity was increased with focal necrosis in hepatic lobules in the transgenic mice later than 24 weeks of age. The transgenic mice expressing osteopontin in hepatocytes may be useful as a model of autoimmune hepatitis. (C) 2004 Elsevier Inc. All rights reserved.