title: High Inter-Individual Diversity of Point Mutations, Insertions, and Deletions in Human Influenza Virus Nucleoprotein-Specific Memory B Cells.
abstract: The diversity of virus-specific antibodies and of B cells among different individuals is unknown. Using single-cell cloning of antibody genes, we generated recombinant human monoclonal antibodies from influenza nucleoprotein-specific memory B cells in four adult humans with and without preceding influenza vaccination. We examined the diversity of the antibody repertoires and found that NP-specific B cells used numerous immunoglobulin genes. The heavy chains (HCs) originated from 26 and the kappa light chains (LCs) from 19 different germ line genes. Matching HC and LC chains gave rise to 43 genetically distinct antibodies that bound influenza NP. The median lengths of the CDR3 of the HC, kappa and lambda LC were 14, 9 and 11 amino acids, respectively. We identified changes at 13.6% of the amino acid positions in the V gene of the antibody heavy chain, at 8.4% in the kappa and at 10.6 % in the lambda V gene. We identified somatic insertions or deletions in 8.1% of the variable genes. We also found several small groups of clonal relatives that were highly diversified. Our findings demonstrate broadly diverse memory B cell repertoires for the influenza nucleoprotein. We found extensive variation within individuals with a high number of point mutations, insertions, and deletions, and extensive clonal diversification. Thus, structurally conserved proteins can elicit broadly diverse and highly mutated B-cell responses.
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Reiche
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany; Department of Biology, Faculty of Natural Sciences, Tishreen University, Latakia, Syria.
authors.forename: Yamen
authors.initials: Y
authors.lastname: Dwai
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany.
authors.forename: Bianca M
authors.initials: BM
authors.lastname: Bussmann
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Horn
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany; Department of Biology, Faculty of Natural Sciences, Tishreen University, Latakia, Syria.
authors.forename: Michael
authors.initials: M
authors.lastname: Sieg
authors.affiliation: Institute of Virology, Faculty of Medicine, University of Leipzig, Leipzig, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Jassoy
mesh_terms: D000328:Adult; D000914:Antibodies, Viral / Q000235:genetics*; D000916:Antibody Diversity / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology*; D019976:Cloning, Organism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007156:Immunologic Memory / Q000235:genetics*; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology; D008297:Male; D016254:Mutagenesis, Insertional / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D019590:Nucleocapsid Proteins; D017354:Point Mutation / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000276:immunology*; D017384:Sequence Deletion / Q000235:genetics*; D014758:Viral Core Proteins / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins