Koch Joachim, Liang Mifang, Queitsch Iris, Kraus Annette A, Bautz Ekkehard K F
Hantavirus-Forschungsstelle der Heidelberger Akademie der Wissenschaften, Im Neuenheimer Feld 230, D-69120 Heidelberg, Germany.
Virology. 2003 Mar 30;308(1):64-73. doi: 10.1016/s0042-6822(02)00094-6.
Old world hantaviruses, causing hemorrhagic fever with renal syndrome (HFRS), still present a public health problem in Asia and Eastern Europe. The majority of cases has been recorded in China. The aim of our study was to generate human recombinant neutralizing antibodies to a hantavirus by phage display technology. To preserve the structural identity of viral protein, the panning procedure was performed on native Hantaan (HTN) (76-118) virus propagated in Vero-E6 cells. In total, five complete human recombinant IgG antibodies were produced in a baculovirus expression system. All of them were able to completely neutralize HTN, and Seoul (SEO) virus in a plaque reduction neutralization test (PRNT). Three of these antibodies could also completely neutralize Dobrava (DOB) virus but not Puumala (PUU) virus. All antibodies bind to Hantaan virus G2 protein localized in the virus envelope. The sequence areas within the HTN (76-118)-G2 protein detected by five selected antibodies were mapped using peptide scans. Two partial epitopes, 916-KVMATIDSF-924 and 954-LVTKDIDFD-963, were recognized, which presumably are of paramount importance for docking of the virus to host cell receptors. A consensus motif 916-KVXATIXSF-924 could be identified by mutational analysis. The neutralizing antibodies to the most widely distributed hantaviruses causing HFRS might be promising candidates for the development of an agent for prevention and treatment of HFRS in patients.
旧世界汉坦病毒可引起肾综合征出血热（HFRS），在亚洲和东欧仍然是一个公共卫生问题。大多数病例在中国被记录。我们研究的目的是通过噬菌体展示技术产生针对汉坦病毒的人重组中和抗体。为了保留病毒蛋白的结构特性，在Vero-E6细胞中繁殖的天然汉滩（HTN）（76-118）病毒上进行淘选程序。总共在杆状病毒表达系统中产生了五种完整的人重组IgG抗体。在蚀斑减少中和试验（PRNT）中，它们都能够完全中和HTN和汉城（SEO）病毒。其中三种抗体也能够完全中和多布拉伐（DOB）病毒，但不能中和普马拉（PUU）病毒。所有抗体都与位于病毒包膜中的汉滩病毒G2蛋白结合。使用肽扫描对五种选定抗体检测到的HTN（76-118）-G2蛋白内的序列区域进行了定位。识别出两个部分表位，916-KVMATIDSF-924和954-LVTKDIDFD-963，推测它们对于病毒与宿主细胞受体的对接至关重要。通过突变分析可以鉴定出一个共有基序916-KVXATIXSF-924。针对引起HFRS的分布最广泛的汉坦病毒的中和抗体可能是开发用于预防和治疗HFRS患者的药物的有前途的候选物。
