title: BCR repertoire sequencing: different patterns of B-cell activation after two Meningococcal vaccines.
abstract: Next-generation sequencing was used to investigate the B-cell receptor heavy chain transcript repertoire of different B-cell subsets (naive, marginal zone (MZ), immunoglobulin M (IgM) memory and IgG memory) at baseline, and of plasma cells (PCs) 7 days following administration of serogroup ACWY meningococcal polysaccharide and protein-polysaccharide conjugate vaccines. Baseline B-cell subsets could be distinguished from each other using a small number of repertoire properties (clonality, mutation from germline and complementarity-determining region 3 (CDR3) length) that were conserved between individuals. However, analyzing the CDR3 amino-acid sequence (which is particularly important for antigen binding) of the baseline subsets showed few sequences shared between individuals. In contrast, day 7 PCs demonstrated nearly 10-fold greater sequence sharing between individuals than the baseline subsets, consistent with the PCs being induced by the vaccine antigen and sharing specificity for a more limited range of epitopes. By annotating PC sequences based on IgG subclass usage and mutation, and also comparing them with the sequences of the baseline cell subsets, we were able to identify different signatures after the polysaccharide and conjugate vaccines. PCs produced after conjugate vaccination were predominantly IgG1, and most related to IgG memory cells. In contrast, after polysaccharide vaccination, the PCs were predominantly IgG2, less mutated and were equally likely to be related to MZ, IgM memory or IgG memory cells. High-throughput B-cell repertoire sequencing thus provides a unique insight into patterns of B-cell activation not possible from more conventional measures of immunogenicity.
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Jacob D
authors.initials: JD
authors.lastname: Galson
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Clutterbuck
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Johannes
authors.initials: J
authors.lastname: Trück
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Maheshi N
authors.initials: MN
authors.lastname: Ramasamy
authors.affiliation: Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
authors.forename: Márton
authors.initials: M
authors.lastname: Münz
authors.affiliation: Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Fowler
authors.affiliation: Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, Oxford, UK.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Cerundolo
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Pollard
authors.affiliation: Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
authors.forename: Gerton
authors.initials: G
authors.lastname: Lunter
authors.affiliation: Oxford Vaccine Group, Department of Paediatrics, University of Oxford and the NIHR Oxford Biomedical Research Center, Oxford, UK.
authors.forename: Dominic F
authors.initials: DF
authors.lastname: Kelly
mesh_terms: D016175:B-Lymphocyte Subsets / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D022801:Complementarity Determining Regions / Q000235:genetics*; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte / Q000378:metabolism; D014644:Genetic Variation / Q000235:genetics; D006684:HLA-DR Antigens / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D007074:Immunoglobulin G / Q000378:metabolism; D007156:Immunologic Memory / Q000235:genetics; D008213:Lymphocyte Activation / Q000235:genetics; D022401:Meningococcal Vaccines / Q000008:administration & dosage / Q000276:immunology*; D025341:Principal Component Analysis; D011947:Receptors, Antigen, B-Cell / Q000235:genetics*; D059467:Transcriptome
chemical_list: D022801:Complementarity Determining Regions; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte; D006684:HLA-DR Antigens; D007074:Immunoglobulin G; D022401:Meningococcal Vaccines; D011947:Receptors, Antigen, B-Cell