title: Single-Cell Analysis of the Plasmablast Response to Vibrio cholerae Demonstrates Expansion of Cross-Reactive Memory B Cells.
abstract: We characterized the acute B cell response in adults with cholera by analyzing the repertoire, specificity, and functional characteristics of 138 monoclonal antibodies (MAbs) generated from single-cell-sorted plasmablasts. We found that the cholera-induced responses were characterized by high levels of somatic hypermutation and large clonal expansions. A majority of the expansions targeted cholera toxin (CT) or lipopolysaccharide (LPS). Using a novel proteomics approach, we were able to identify sialidase as another major antigen targeted by the antibody response to Vibrio cholerae infection. Antitoxin MAbs targeted both the A and B subunits, and most were also potent neutralizers of enterotoxigenic Escherichia coli heat-labile toxin. LPS-specific MAbs uniformly targeted the O-specific polysaccharide, with no detectable responses to either the core or the lipid moiety of LPS. Interestingly, the LPS-specific antibodies varied widely in serotype specificity and functional characteristics. One participant infected with the Ogawa serotype produced highly mutated LPS-specific antibodies that preferentially bound the previously circulating Inaba serotype. This demonstrates durable memory against a polysaccharide antigen presented at the mucosal surface and provides a mechanism for the long-term, partial heterotypic immunity seen following cholera.
  
  
  
  Cholera is a diarrheal disease that results in significant mortality. While oral cholera vaccines are beneficial, they do not achieve equivalent protection compared to infection with Vibrio cholerae Although antibodies likely mediate protection, the mechanisms of immunity following cholera are poorly understood, and a detailed understanding of antibody responses to cholera is of significance for human health. In this study, we characterized the human response to cholera at the single-plasmablast, monoclonal antibody level. Although this approach has not been widely applied to the study of human bacterial infection, we were able to uncover the basis of cross-reactivity between different V. cholerae serotypes and the likely impact of prior enterotoxigenic Escherichia coli exposure on the response to cholera, as well as identify novel antigenic targets. In addition to improving our understanding of the repertoire and function of the antibody response to cholera in humans, this study has implications for future cholera vaccination efforts.
authors.affiliation: Division of Infectious Disease, Department of Pediatrics, Emory University, School of Medicine, Atlanta, Georgia, USA.|Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Kauffman
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Taufiqur R
authors.initials: TR
authors.lastname: Bhuiyan
authors.affiliation: Department of Medicine, University of California, Irvine, California, USA.
authors.forename: Rie
authors.initials: R
authors.lastname: Nakajima
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Leslie M
authors.initials: LM
authors.lastname: Mayo-Smith
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Rasheduzzaman
authors.initials: R
authors.lastname: Rashu
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Mohammad Rubel
authors.initials: MR
authors.lastname: Hoq
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Fahima
authors.initials: F
authors.lastname: Chowdhury
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Ashraful Islam
authors.initials: AI
authors.lastname: Khan
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.|Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Atiqur
authors.initials: A
authors.lastname: Rahman
authors.affiliation: Division of Infectious Disease, Department of Pediatrics, Emory University, School of Medicine, Atlanta, Georgia, USA.|Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Siddhartha K
authors.initials: SK
authors.lastname: Bhaumik
authors.affiliation: Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Levelle
authors.initials: L
authors.lastname: Harris
authors.affiliation: Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Justin T
authors.initials: JT
authors.lastname: O'Neal
authors.affiliation: Division of Infectious Disease, Department of Pediatrics, Emory University, School of Medicine, Atlanta, Georgia, USA.|Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Jessica F
authors.initials: JF
authors.lastname: Trost
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Nur Haq
authors.initials: NH
authors.lastname: Alam
authors.affiliation: Department of Medicine, University of California, Irvine, California, USA.
authors.forename: Algis
authors.initials: A
authors.lastname: Jasinskas
authors.affiliation: Department of Medicine, University of California, Irvine, California, USA.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Dotsey
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Meagan
authors.initials: M
authors.lastname: Kelly
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Richelle C
authors.initials: RC
authors.lastname: Charles
authors.affiliation: National Institute of Diabetes and Digestive and Kidney Diseases, Laboratory of Bioorganic Chemistry, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.affiliation: National Institute of Diabetes and Digestive and Kidney Diseases, Laboratory of Bioorganic Chemistry, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Pavol
authors.initials: P
authors.lastname: Kováč
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Calderwood
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.|Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA.
authors.forename: Edward T
authors.initials: ET
authors.lastname: Ryan
authors.affiliation: Department of Medicine, University of California, Irvine, California, USA.
authors.forename: Phillip L
authors.initials: PL
authors.lastname: Felgner
authors.affiliation: Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Firdausi
authors.initials: F
authors.lastname: Qadri
authors.affiliation: Division of Infectious Disease, Department of Pediatrics, Emory University, School of Medicine, Atlanta, Georgia, USA jwramme@emory.edu jbharris@partners.org.|Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Wrammert
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA jwramme@emory.edu jbharris@partners.org.|Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Jason B
authors.initials: JB
authors.lastname: Harris
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology*; D001427:Bacterial Toxins / Q000276:immunology; D002771:Cholera / Q000276:immunology* / Q000382:microbiology; D002772:Cholera Toxin / Q000276:immunology; D003429:Cross Reactions; D004768:Enterotoxins / Q000276:immunology; D029968:Escherichia coli Proteins / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology; D009439:Neuraminidase / Q000276:immunology; D019081:O Antigens / Q000276:immunology; D040901:Proteomics / Q000379:methods; D065288:Serogroup; D059010:Single-Cell Analysis; D014734:Vibrio cholerae / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D019081:O Antigens; D002772:Cholera Toxin; C027538:heat-labile enterotoxin, E coli; D009439:Neuraminidase