Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 02139.
Proc Natl Acad Sci U S A. 2014 Apr 1;111(13):4928-33. doi: 10.1073/pnas.1323862111. Epub 2014 Mar 17.
The adaptive immune system confers protection by generating a diverse repertoire of antibody receptors that are rapidly expanded and contracted in response to specific targets. Next-generation DNA sequencing now provides the opportunity to survey this complex and vast repertoire. In the present work, we describe a set of tools for the analysis of antibody repertoires and their application to elucidating the dynamics of the response to viral vaccination in human volunteers. By analyzing data from 38 separate blood samples across 2 y, we found that the use of the germ-line library of V and J segments is conserved between individuals over time. Surprisingly, there appeared to be no correlation between the use level of a particular VJ combination and degree of expansion. We found the antibody RNA repertoire in each volunteer to be highly dynamic, with each individual displaying qualitatively different response dynamics. By using combinatorial phage display, we screened selected VH genes paired with their corresponding VL library for affinity against the vaccine antigens. Altogether, this work presents an additional set of tools for profiling the human antibody repertoire and demonstrates characterization of the fast repertoire dynamics through time in multiple individuals responding to an immune challenge.
适应性免疫系统通过生成多样化的抗体受体库来提供保护，这些受体库可以针对特定目标迅速扩增和收缩。新一代 DNA 测序现在提供了调查这种复杂而广泛的受体库的机会。在本工作中，我们描述了一套用于分析抗体受体库的工具，并将其应用于阐明人类志愿者对病毒疫苗接种反应的动态。通过分析 2 年内 38 个单独血液样本的数据，我们发现个体之间的 V 和 J 节段的种系库的使用在时间上是保守的。令人惊讶的是，特定 VJ 组合的使用水平与扩增程度之间似乎没有相关性。我们发现每个志愿者的抗体 RNA 受体库都是高度动态的，每个个体显示出定性不同的反应动力学。通过使用组合噬菌体展示，我们筛选了与相应 VL 文库配对的选定 VH 基因，以检测它们对疫苗抗原的亲和力。总的来说，这项工作提供了一组用于分析人类抗体库的工具，并通过对多个个体对免疫挑战的反应进行时间上的分析，展示了快速受体库动力学的特征。
