Generation of recombinant vaccinia viruses expressing Puumala virus proteins and use in isolating cytotoxic T cells specific for Puumala virus
Generation of recombinant vaccinia viruses expressing Puumala virus proteins and use in isolating cytotoxic T cells specific for Puumala virus
复制标题
DOI：
10.1016/s0168-1702(01)00416-6
复制
发表时间：
2002-03-20
期刊：
影响因子：
5
通讯作者：
Ennis, FA
中科院分区：
文献类型：
--
作者：
Terajima, M;Van Epps, HL;Ennis, FA
普玛拉(PUU)病毒在欧洲引起一种形式的肾综合征出血热(HFRS)，称为流行性肾病(NE)。HFRS的特点是毛细血管通透性增加，我们推测这是由宿主免疫系统，特别是细胞免疫反应过度激活引起的。为了鉴定针对NE患者PUU病毒的细胞毒性T淋巴细胞(CTL)，我们制备了表达PUU病毒蛋白、核衣壳蛋白(N)和两种表面糖蛋白G1和G2的重组痘苗病毒。在强合成启动子的控制下，获得了携带N或G2基因前半部分的重组痘苗病毒。然而，为了表达G1和G2蛋白的后半部分，我们需要使用T7表达系统，其中T7RNA聚合酶是由另一种重组痘苗病毒共同感染相同的细胞产生的。用这些重组痘苗病毒从NE患者外周血单个核细胞中检测和克隆PUU病毒特异性CTL。筛选出一株识别G2蛋白的人类白细胞抗原A24限制性CTL株，并测定了其9聚体表位。(C)2002 Elsevier Science B.V.保留所有权利。
Puumala (PUU) virus causes a form of hemorrhagic fever with renal syndrome (HFRS), called nephropathia epidemica (NE), in Europe. HFRS is characterized by an increased capillary permeability, which we hypothesize is caused by hyperactivation of the host immune system, especially cellular immune responses. To identify cytotoxic T lymphocytes (CTLs) specific for the PUU virus from NE patients, we have made recombinant vaccinia viruses expressing PUU virus proteins, the nucleocapsid (N) and two surface glycoproteins, G1 and G2. Recombinant vaccinia viruses carrying the N or the first half of the G2 cDNA under the control of a strong synthetic promoter were made. To express G1 and the second half of the G2 proteins, however, we needed to use a T7 expression system, where the T7 RNA polymerase is produced from another recombinant vaccina virus co-infecting the same cells. These recombinant vaccinia viruses were used to detect and clone PUU virus-specific CTLs from the peripheral blood mononuclear cells of NE patients. An HLA-A24-restricted CTL line recognizing the G2 protein was isolated and its 9-mer epitope was determined. (C) 2002 Elsevier Science B.V. All rights reserved.