NMO sera down-regulate AQP4 in human astrocyte and induce cytotoxicity independent of complement
NMO sera down-regulate AQP4 in human astrocyte and induce cytotoxicity independent of complement
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DOI：
10.1016/j.jns.2013.05.035
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发表时间：
2013-08-15
影响因子：
4.4
通讯作者：
Kanda, Takashi
中科院分区：
文献类型：
--
作者：
Haruki, Hiroyo;Sano, Yasuteru;Kanda, Takashi
星形胶质细胞水通道水通道蛋白 4 (AQP4) 自身抗体对视神经脊髓炎 (NMO) 具有高度特异性。然而，NMO的分子机制仍不清楚。本研究的目的是确定导致星形胶质细胞损伤发生的可能的体液机制。人原代星形胶质细胞（AST）通过含有温度敏感的SV40 T抗原基因和AQP4 cDNA（M23）的逆转录病毒载体永生化，命名为hAST-AQP4。评估了 NMO 血清对 AQP4 含量和定位的影响，包括细胞毒性和星形胶质细胞形态。此外，本研究还检验了星形胶质细胞中 AQP4 蛋白的数量和定位是否受到与永生化人脑微血管内皮细胞系 TY09 直接接触的影响。单独的 NMO 血清诱导细胞毒性，并且添加补体对 hAST-AQP4 具有更大的有害影响。 NMO 血清还降低了 AQP4 mRNA 和蛋白。单独的 NMO 血清上调星形胶质细胞中的 TNF α 和 IL-6，与抗 TNF α 和抗 1L-6 中和抗体共孵育可阻断星形胶质细胞中的细胞毒性和 AQP4 的减少。在使用体外BBB模型的实验中，与TY09共培养后，AQP4蛋白主要定位于星形胶质细胞膜上，与TY09接触。因此，未来阐明上调星形胶质细胞中 AQP4 的因子（推测是由血脑屏障形成内皮细胞释放的）并阻止炎症细胞因子的产生，可能会导致新治疗策略的开发。 (C) 2013 Elsevier B.V. 保留所有权利。
Autoantibodies against astrocyte water channel aquaporin-4 (AQP4) are highly specific for neuromyelitis optica (NMO). However, the molecular mechanism of NMO still remains unclear.The purpose of this study was to identify the possible humoral mechanisms responsible for the occurrence of astrocytic damage. Human primary astrocytes (AST) were immortalized by retroviral vectors harboring temperature-sensitive SV40 T antigen gene and AQP4 cDNA (M23), designated as hAST-AQP4. The effects of NMO sera on the content and localization of AQP4, including cytotoxicity and astrocytic morphology, were evaluated. In addition, this study examined whether the amount and localization of AQP4 protein in astrocytes were influenced by direct contact with the immortalized human brain microvascular endothelial cell line, TY09. NMO sera alone induced cytotoxicity and addition of complement had a more harmful effect on hAST-AQP4. NMO sera also decreased AQP4 mRNA and protein. NMO sera alone up-regulated TNF alpha and IL-6 in astrocytes and co-incubation with anti-TNF alpha and anti-1L-6 neutralizing antibodies blocked both the cytotoxicity and reduction of AQP4 in astrocytes. In the experiment using the in vitro BBB models, AQP4 protein mainly localized at the astrocytic membrane after co-culture with TY09, in contact with TY09. The future elucidation of factors that up-regulate AQP4 in astrocytes presumably released by blood brain barrier forming endothelial cells and that block the production of inflammatory cytokines may therefore lead to the development of a novel therapeutic strategy. (C) 2013 Elsevier B.V. All rights reserved.