Cloning the antibody response in humans with inflammatory CNS disease: isolation of measles virus-specific antibodies from phage display libraries of a subacute sclerosing panencephalitis brain
Cloning the antibody response in humans with inflammatory CNS disease: isolation of measles virus-specific antibodies from phage display libraries of a subacute sclerosing panencephalitis brain
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DOI：
10.1016/s0165-5728(98)00243-4
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发表时间：
1999-02-01
影响因子：
3.3
通讯作者：
Gilden, DH
中科院分区：
文献类型：
--
作者：
Burgoon, MP;Williamson, RA;Gilden, DH
我们已经开发出一种策略，以确定在慢性炎症性中枢神经系统疾病表现出鞘内表达的寡克隆IgG的疾病相关抗原。以麻疹病毒感染的亚急性硬化性全脑炎（SSPE）为模型系统，从脑组织中表达的IgG扩增产物中构建噬菌体展示抗体Fab库。对麻疹病毒感染的细胞裂解物的库的选择产生了四个不同的Fab，通过ELISA和免疫染色，特异性地与麻疹病毒感染的细胞反应。三个Fab免疫沉淀72 kDa的蛋白质从感染的细胞培养物对应的麻疹病毒磷蛋白。第四个Fab免疫沉淀并通过免疫印迹识别对应于麻疹病毒核蛋白的60 kDa蛋白。结果表明，来自炎性CNS疾病的功能性抗体可以在细菌中表达并用于鉴定疾病相关抗原。这种方法可以应用于不明原因的慢性炎症性CNS疾病，如多发性硬化症。(C)1999 Elsevier Science B.V.保留所有权利。
We have developed a strategy to identify the disease-relevant antigens in a chronic inflammatory CNS disease exhibiting intrathecally expressed oligoclonal IgG. Using subacute sclerosing panencephalitis (SSPE), a chronic inflammatory measles virus infection of the brain as a model system, we constructed a phage display antibody Fab Library from the amplified products of IgG expressed in the brain. Selection of the library against measles virus-infected cell lysates yielded four distinct Fabs which, by ELISA and by immunostaining, reacted specifically with measles virus-infected cells. Three Fabs immunoprecipitated a 72 kDa protein from infected cell cultures corresponding to the measles virus phosphoprotein. The fourth Fab immunoprecipitated and recognized by immunoblotting a 60 kDa protein corresponding to the measles virus nucleoprotein. The results demonstrate that functional antibodies from an inflammatory CNS disease can be expressed in bacteria and used to identify disease-relevant antigens. This approach could be applied to chronic inflammatory CNS diseases of unknown cause such as multiple sclerosis. (C) 1999 Elsevier Science B.V. All rights reserved.