Rapid cloning of high-affinity human monoclonal antibodies against influenza virus
Rapid cloning of high-affinity human monoclonal antibodies against influenza virus
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DOI：
10.1038/nature06890
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发表时间：
2008-05-29
期刊：
影响因子：
64.8
通讯作者：
Wilson, Patrick C.
中科院分区：
文献类型：
--
作者：
Wrammert, Jens;Smith, Kenneth;Wilson, Patrick C.
预先存在的中和抗体通常是抵御病原体的第一道防线。对于流感病毒，每年都会进行疫苗接种，以维持针对当前流行毒株的抗体保护水平。在此我们报道，加强免疫接种后，会出现快速且强烈的流感特异性IgG(+)抗体分泌浆细胞（ASC）反应，该反应在大约第7天达到峰值，占外周血B细胞的比例高达6%。这些ASC可与流感特异性IgG(+)记忆B细胞区分开来，记忆B细胞在接种疫苗后14 - 21天达到峰值，平均占所有B细胞的1%。重要的是，在反应峰值时纯化的ASC中多达80%是流感特异性的。这种ASC反应的特征是B细胞受体（BCR）库高度受限，在一些供体中仅由少数几个B细胞克隆主导。然而，这种寡克隆反应显示出因体细胞突变积累而产生的广泛的克隆内多样化。我们利用从分选的单个ASC中分离出的免疫球蛋白可变区，生产了50多种人单克隆抗体（mAbs），这些抗体以高亲和力与三种流感疫苗毒株结合。这一策略表明，我们能够在接种疫苗后的一个月内从人体中产生多种高亲和力的mAbs。这组流感病毒特异性人单克隆抗体使我们能够探讨原始抗原痕迹（OAS）问题：即诱导产生的抗体与疫苗中存在的病毒毒株相比，对先前接触过的流感病毒毒株表现出更高亲和力的现象(1)。然而，我们发现大多数流感病毒特异性mAbs对当前疫苗毒株表现出最高亲和力。因此，在接受流感疫苗接种的正常、健康成年人中，OAS似乎并不常见。
Pre- existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG(+) antibody- secreting plasma cell ( ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza- specific IgG(+) memory B cells that peaked 14 - 21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B- cell receptor ( BCR) repertoire that in some donors was dominated by only a few B- cell clones. This pauciclonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies ( mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza- virus- specific human mAbs allowed us to address the issue of original antigenic sin ( OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine(1). However, we found that most of the influenza- virus- specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.