Recombinant protein comprising multi-neutralizing epitopes induced high titer of antibodies against Influenza A virus
Recombinant protein comprising multi-neutralizing epitopes induced high titer of antibodies against Influenza A virus
复制标题
DOI：
10.1078/0171-2985-00244
复制
发表时间：
2003-01-01
期刊：
影响因子：
2.8
通讯作者：
Chen, YH
中科院分区：
文献类型：
--
作者：
Li, H;Ding, J;Chen, YH
在以往的研究中，我们认为表位疫苗可能是一种新的抗病毒策略。基于这一假设，我们设计并表达了一个包含H3 N2流感病毒血凝素（HA）三个中和表位（aa 92 -105、127-133和183-195）重复序列的重组免疫原（多表位肽）。杆菌免疫小鼠和家兔后，重组多表位蛋白均能诱导产生高水平的多表位特异性抗体。用三种不同的表位肽（合成肽）在ELISA法中检测血清中的表位特异性抗体，并确认血清稀释度为1：6400至1：25600。Western blot分析表明，抗血清和经合成表位-肽偶联的Sepharose柱纯化的抗体均能识别流感病毒（A/Wuhan/359/95 H3 N2株）的天然HA。在血凝抑制（HI）试验中，这三种抗血清在1：20至1：80的稀释度下显示抑制活性。有趣的是，由表位疫苗诱导的抗血清和纯化抗体可部分抑制流感病毒（A/Wuhan/359/95株）在MDCK细胞单层上的空斑形成。这些结果表明，重组多表位疫苗能够同时诱导多种抗流感病毒活性，这可能为研制有效的流感病毒疫苗提供新的途径。
In previous studies, we suggested that epitope-vaccine might be a new strategy against virus infection. Based on this hypothesis, we designed and expressed a recombinant immunogen (multi-epitope-peptide) comprising repeats of three neutralizing-epitopes (neutralizing epitopes: aa92-105, 127-133 and 183-195) of hemagglutininin (HA) of influenza virus (H3N2) in E. coli. After vaccination, the recombinant multi-epitope protein could induce a high level of antibodies with predefined multi-epitope-specificity in mice and rabbits. The epitope-specific antibodies in sera were tested using three different epitope-peptides (synthetic peptides) in ELISA assay, and the serum dilutions from 1:6400 to 1:25600 were confirmed. In western blot analysis, both the antiserum and the antibodies purified by synthetic epitope-peptide coupled sepharose columns could recognize natural HA from influenza virus particles (strain A/Wuhan/359/95 H3N2). In hemagglutination inhibition (HI) tests, these three antisera at the dilutions from 1:20 to 1 :80 showed inhibitory activity. Interestingly, antisera and purified antibodies induced by the epitope-vaccine could partially inhibit plaque-formation of influenza virus (strain A/Wuhan/359/95) on MDCK cell monolayers. These results suggest that the recombinant multi-epitope vaccine can simultaneously induce multi-antiviral activities against influenza virus, which may provide a new way to develop effective vaccines against influenza virus.