In vitro evolution of an influenza broadly neutralizing antibody is modulated by hemagglutinin receptor specificity.
In vitro evolution of an influenza broadly neutralizing antibody is modulated by hemagglutinin receptor specificity.
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DOI：
10.1038/ncomms15371
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发表时间：
2017-05-15
影响因子：
16.6
通讯作者：
Wilson IA
中科院分区：
文献类型：
--
作者：
Wu NC;Grande G;Turner HL;Ward AB;Xie J;Lerner RA;Wilson IA
针对流感病毒的人广泛中和抗体（bnAb）的相对新近的发现和表征为抗病毒和疫苗开发提供了有价值的见解。然而，影响高亲和力bnAb进化的因素仍然难以捉摸。因此，我们探索bnAb C05的功能序列空间，其通过长CDR H3靶向流感血凝素（HA）的受体结合位点（RBS）。我们将联合收割机饱和诱变与酵母展示相结合，以富集与H1和H3 HA结合的CDR H3的C05变体。C05变体进化出高达20倍的亲和力，但增加了对选择中使用的每种HA亚型的特异性。结构分析表明，精细的特异性受到高度保守的取代的强烈影响，该取代调节不同亚型中的受体结合。总的来说，这项研究表明，亚型和物种之间的HA RBS的微妙的自然变化可能会不同地影响高亲和力bnAb的进化。针对流感血凝素（HA）的广泛中和抗体（bnAb）已经为抗病毒开发提供了见解。在这里，作者采用饱和诱变的互补位区的bnAb结合酵母展示筛选使用H1和H3 HA，并发现之间的权衡存在抗体亲和力和宽度，受不同的受体结合模式的影响。
The relatively recent discovery and characterization of human broadly neutralizing antibodies (bnAbs) against influenza virus provide valuable insights into antiviral and vaccine development. However, the factors that influence the evolution of high-affinity bnAbs remain elusive. We therefore explore the functional sequence space of bnAb C05, which targets the receptor-binding site (RBS) of influenza haemagglutinin (HA) via a long CDR H3. We combine saturation mutagenesis with yeast display to enrich for C05 variants of CDR H3 that bind to H1 and H3 HAs. The C05 variants evolve up to 20-fold higher affinity but increase specificity to each HA subtype used in the selection. Structural analysis reveals that the fine specificity is strongly influenced by a highly conserved substitution that regulates receptor binding in different subtypes. Overall, this study suggests that subtle natural variations in the HA RBS between subtypes and species may differentially influence the evolution of high-affinity bnAbs. Broadly neutralizing antibodies (bnAbs) against influenza hemagglutinin (HA) have yielded insights for antiviral development. Here, the authors employ saturated mutagenesis of the paratope region of a bnAb combined with yeast display screening using H1 and H3 HAs, and find that a tradeoff exists between Ab affinity and breadth that influenced by disparate modes of receptor binding.