Dai Kaifan, He Linling, Khan Salar N, O'Dell Sijy, McKee Krisha, Tran Karen, Li Yuxing, Sundling Christopher, Morris Charles D, Mascola John R, Karlsson Hedestam Gunilla B, Wyatt Richard T, Zhu Jiang
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
Vaccine Research Center, National Institutes of Health, Bethesda, Maryland, USA.
mBio. 2015 Nov 3;6(6):e01375-15. doi: 10.1128/mBio.01375-15.
Next-generation sequencing (NGS) has been used to investigate the diversity and maturation of broadly neutralizing antibodies (bNAbs) in HIV-1-infected individuals. However, the application of NGS to the preclinical assessment of human vaccines, particularly the monitoring of vaccine-induced B-cell responses in a nonhuman primate (NHP) model, has not been reported. Here, we present a longitudinal NGS analysis of memory B-cell responses to an HIV-1 trimer vaccine in a macaque that has been extensively studied by single B-cell sorting and antibody characterization. We first established an NHP antibodyomics pipeline using the available 454 pyrosequencing data from this macaque and developed a protocol to sequence the NHP antibody repertoire in an unbiased manner. Using these methods, we then analyzed memory B-cell repertoires at four time points of NHP immunization and traced the lineages of seven CD4-binding site (CD4bs)-directed monoclonal antibodies previously isolated from this macaque. Longitudinal analysis revealed distinct patterns of B-cell lineage development in response to an HIV-1 trimer vaccine. While the temporal B-cell repertoire profiles and lineage patterns provide a baseline for comparison with forthcoming HIV-1 trimer vaccines, the newly developed NHP antibody NGS technologies and antibodyomics tools will facilitate future evaluation of human vaccine candidates.
The nonhuman primate model has been widely used in the preclinical assessment of human vaccines. Next-generation sequencing of B-cell repertoires provides a quantitative tool to analyze B-cell responses to a vaccine. In this study, the longitudinal B-cell repertoire analysis of a rhesus macaque immunized with an HIV-1 trimer vaccine revealed complex B-cell lineage patterns and showed the potential to facilitate the evaluation of future HIV-1 vaccines. The repertoire sequencing technologies and antibodyomics methods reported here can be extended to vaccine development for other human pathogens utilizing the nonhuman primate model.
下一代测序（NGS）已被用于研究HIV-1感染个体中广泛中和抗体（bNAb）的多样性和成熟情况。然而，NGS在人类疫苗临床前评估中的应用，特别是在非人类灵长类动物（NHP）模型中监测疫苗诱导的B细胞反应，尚未见报道。在此，我们对一只猕猴针对HIV-1三聚体疫苗的记忆B细胞反应进行了纵向NGS分析，该猕猴已通过单细胞B细胞分选和抗体表征进行了广泛研究。我们首先利用该猕猴现有的454焦磷酸测序数据建立了一个NHP抗体组学流程，并开发了一种以无偏方式对NHP抗体库进行测序的方案。然后，使用这些方法，我们分析了NHP免疫四个时间点的记忆B细胞库，并追踪了先前从该猕猴分离出的七种靶向CD4结合位点（CD4bs）的单克隆抗体的谱系。纵向分析揭示了针对HIV-1三聚体疫苗的B细胞谱系发育的不同模式。虽然B细胞库的时间谱和谱系模式为与即将推出的HIV-1三聚体疫苗进行比较提供了基线，但新开发的NHP抗体NGS技术和抗体组学工具将有助于未来对人类疫苗候选物的评估。
非人类灵长类动物模型已广泛用于人类疫苗的临床前评估。B细胞库的下一代测序提供了一种定量工具来分析B细胞对疫苗的反应。在本研究中，对用HIV-1三聚体疫苗免疫的恒河猴进行的纵向B细胞库分析揭示了复杂的B细胞谱系模式，并显示出促进未来HIV-1疫苗评估的潜力。此处报道的库测序技术和抗体组学方法可扩展到利用非人类灵长类动物模型开发针对其他人类病原体的疫苗。
