Molecular cloning, expression and characterization of the functional domain of CTLA4 from the rhesus monkey, Macaca mulatta
Molecular cloning, expression and characterization of the functional domain of CTLA4 from the rhesus monkey, Macaca mulatta
复制标题
恒河猴 CTLA4 功能域的分子克隆、表达和表征。
DOI：
10.1016/j.dci.2011.02.007
复制
发表时间：
2011-07-01
影响因子：
2.9
通讯作者：
Lu, Xiaofeng
中科院分区：
文献类型：
--
作者：
Zhu, Shengyun;Liu, Shan;Lu, Xiaofeng
细胞毒性T淋巴细胞相关抗原4(CTLA4)是一种有效的T细胞活化抑制因子。从外周血单个核细胞(PBMC)中分离到猕猴CTLA4(MmCTLA4)膜蛋白和可溶性蛋白的编码基因。预测的MmCTLA4蛋白与人CTLA4(HCTLA4)几乎相同，除了第49位的丝氨酸而不是天冬酰胺残基和141位的亮氨酸而不是蛋氨酸残基。在毕赤酵母中表达了融合蛋白MmCTLA4Ig，该融合蛋白含有MmCTLA4的胞外区和人IgG1抗体的恒定区。巴斯德毕赤酵母产生的mCTLA4Ig与人B7阳性Raji细胞具有特异性结合，这种结合可被hCTLA4Ig竞争结合所抑制。MmCTLA4Ig和hCTLA4Ig对刀豆蛋白A(ConA)或同种异体细胞刺激的恒河猴、人或小鼠淋巴细胞的增殖有相似的抑制作用。这些结果表明，MmCTLA4是T细胞活化的负性调节因子，MmCTLA4Ig可能用于猕猴免疫性疾病的免疫治疗。(C)2011爱思唯尔有限公司。保留所有权利。
Cytotoxic T lymphocyte-associated antigen 4 (CTLA4) is a potent inhibitor of T cell activation. The genes encoding the membrane and soluble forms of Macaca mulatto CTLA4 (mmCTLA4) were isolated from peripheral blood mononuclear cells (PBMCs). The predicted mmCTLA4 protein is nearly identical to human CTLA4 (hCTLA4), with the exception of a serine instead of an asparagine residue at position 49 and a leucine instead of a methionine residue at position 141 of its extracellular domain. The fusion protein mmCTLA4Ig, containing the extracellular domain of mmCTLA4 and the constant region of the human IgG1 antibody, was expressed in Pichia pastoris. The mmCTLA4Ig produced by P. pastoris exhibited specific binding to human B7-positive Raji cells that could be inhibited by competitive binding of hCTLA4Ig. MmCTLA4Ig and hCTLA4Ig could comparably suppress the proliferation of lymphocytes derived from rhesus monkeys, humans, or mice that had been stimulated either by concanavalin A (Con A) or allogeneic cells. These results suggest that mmCTLA4 is a negative regulator of T cell activation and that mmCTLA4Ig may be useful for immunotherapy of immunologic diseases in the rhesus monkey. (C) 2011 Elsevier Ltd. All rights reserved.