title: Rapid cloning of high-affinity human monoclonal antibodies against influenza virus.
abstract: Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14-21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.
authors.affiliation: Emory Vaccine Center and Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Wrammert
authors.forename: Kenneth
authors.initials: K
authors.lastname: Smith
authors.forename: Joe
authors.initials: J
authors.lastname: Miller
authors.forename: William A
authors.initials: WA
authors.lastname: Langley
authors.forename: Kenneth
authors.initials: K
authors.lastname: Kokko
authors.forename: Christian
authors.initials: C
authors.lastname: Larsen
authors.forename: Nai-Ying
authors.initials: NY
authors.lastname: Zheng
authors.forename: Israel
authors.initials: I
authors.lastname: Mays
authors.forename: Lori
authors.initials: L
authors.lastname: Garman
authors.forename: Christina
authors.initials: C
authors.lastname: Helms
authors.forename: Judith
authors.initials: J
authors.lastname: James
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Air
authors.forename: J Donald
authors.initials: JD
authors.lastname: Capra
authors.forename: Rafi
authors.initials: R
authors.lastname: Ahmed
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Wilson
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D002999:Clone Cells / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D007117:Immunization, Secondary; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D007156:Immunologic Memory / Q000276:immunology*; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology; D009981:Influenza B virus / Q000276:immunology; D007252:Influenza Vaccines / Q000276:immunology; D018448:Models, Immunological; D009975:Orthomyxoviridae / Q000276:immunology*; D010950:Plasma Cells / Q000276:immunology / Q000378:metabolism; D027041:Somatic Hypermutation, Immunoglobulin / Q000235:genetics; D013997:Time Factors; D014611:Vaccination
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007252:Influenza Vaccines