title: Rhesus Macaque B-Cell Responses to an HIV-1 Trimer Vaccine Revealed by Unbiased Longitudinal Repertoire Analysis.
abstract: 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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Kaifan
authors.initials: K
authors.lastname: Dai
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Linling
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Salar N
authors.initials: SN
authors.lastname: Khan
authors.affiliation: Vaccine Research Center, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Sijy
authors.initials: S
authors.lastname: O'Dell
authors.affiliation: Vaccine Research Center, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Krisha
authors.initials: K
authors.lastname: McKee
authors.affiliation: IAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Tran
authors.affiliation: Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, Maryland, USA.
authors.forename: Yuxing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Microbiology and Tumor Cell Biology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Christopher
authors.initials: C
authors.lastname: Sundling
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Charles D
authors.initials: CD
authors.lastname: Morris
authors.affiliation: Vaccine Research Center, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Mascola
authors.affiliation: Department of Microbiology and Tumor Cell Biology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Gunilla B
authors.initials: GB
authors.lastname: Karlsson Hedestam
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA IAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, California, USA Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, California, USA wyatt@scripps.edu jiang@scripps.edu.
authors.forename: Richard T
authors.initials: RT
authors.lastname: Wyatt
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, California, USA Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, USA wyatt@scripps.edu jiang@scripps.edu.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhu
mesh_terms: D016915:AIDS Vaccines / Q000008:administration & dosage / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D001402:B-Lymphocytes / Q000145:classification / Q000276:immunology*; D001666:Binding Sites, Antibody; D019070:Cell Lineage; D019295:Computational Biology; D030541:Databases, Genetic; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D015497:HIV-1 / Q000276:immunology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007156:Immunologic Memory; D008253:Macaca mulatta; D017422:Sequence Analysis, DNA; D014611:Vaccination
chemical_list: D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D015483:HIV Antibodies