title: Mapping the Human Memory B Cell and Serum Neutralizing Antibody Responses to Dengue Virus Serotype 4 Infection and Vaccination.
abstract: The four dengue virus (DENV) serotypes are mosquito-borne flaviviruses responsible for dengue fever and dengue hemorrhagic fever. People exposed to DENV develop antibodies (Abs) that strongly neutralize the serotype responsible for infection. Historically, infection with DENV serotype 4 (DENV4) has been less common and less studied than infections with the other three serotypes. However, DENV4 has been responsible for recent large and sustained epidemics in Asia and Latin America. The neutralizing antibody responses and the epitopes targeted against DENV4 have not been characterized in human infection. In this study, we mapped and characterized epitopes on DENV4 recognized by neutralizing antibodies in people previously exposed to DENV4 infections or to a live attenuated DENV4 vaccine. To study the fine specificity of DENV4 neutralizing human antibodies, B cells from two people exposed to DENV4 were immortalized and screened to identify DENV-specific clones. Two human monoclonal antibodies (MAbs) that neutralized DENV4 were isolated, and their epitopes were finely mapped using recombinant viruses and alanine scan mutation array techniques. Both antibodies bound to quaternary structure epitopes near the hinge region between envelope protein domain I (EDI) and EDII. In parallel, to characterize the serum neutralizing antibody responses, convalescence-phase serum samples from people previously exposed to primary DENV4 natural infections or a monovalent DENV4 vaccine were analyzed. Natural infection and vaccination also induced serum-neutralizing antibodies that targeted similar epitope domains at the EDI/II hinge region. These studies defined a target of neutralizing antigenic site on DENV4 targeted by human antibodies following natural infection or vaccination.IMPORTANCEThe four serotypes of dengue virus are the causative agents of dengue fever and dengue hemorrhagic fever. People exposed to primary DENV infections develop long-term neutralizing antibody responses, but these principally recognize only the infecting serotype. An effective vaccine against dengue should elicit long-lasting protective antibody responses to all four serotypes simultaneously. We and others have defined antigenic sites on the envelope (E) protein of viruses of dengue virus serotypes 1, 2, and 3 targeted by human neutralizing antibodies. The epitopes on DENV4 E protein targeted by the human neutralizing antibodies and the mechanisms of serotype 4 neutralization are poorly understood. Here, we report the properties of human antibodies that neutralize dengue virus serotype 4. People exposed to serotype 4 infections or a live attenuated serotype 4 vaccine developed neutralizing antibodies that bound to similar sites on the viral E protein. These studies have provided a foundation for developing and evaluating DENV4 vaccines.
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Usha K
authors.initials: UK
authors.lastname: Nivarthi
authors.affiliation: The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
authors.forename: Nurgun
authors.initials: N
authors.lastname: Kose
authors.affiliation: The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA.|Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
authors.forename: Gopal
authors.initials: G
authors.lastname: Sapparapu
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: Widman
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Gallichotte
authors.affiliation: Integral Molecular Inc., Philadelphia, Pennsylvania, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Pfaff
authors.affiliation: Integral Molecular Inc., Philadelphia, Pennsylvania, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Doranz
authors.affiliation: La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.
authors.forename: Daniela
authors.initials: D
authors.lastname: Weiskopf
authors.affiliation: La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sette
authors.affiliation: Johns Hopkins Bloomberg School of Public Health, Department of International Health, Baltimore, Maryland, USA.|Center for Immunization Research, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA.
authors.forename: Anna P
authors.initials: AP
authors.lastname: Durbin
authors.affiliation: Laboratory of Infectious Diseases, NIAID, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Steve S
authors.initials: SS
authors.lastname: Whitehead
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
authors.forename: Ralph
authors.initials: R
authors.lastname: Baric
authors.affiliation: The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA james.crowe@vanderbilt.edu aravinda_desilva@med.unc.edu.|Department of Pediatrics, Vanderbilt University Medical Center, Nashville, Tennessee, USA.|Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Crowe
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA james.crowe@vanderbilt.edu aravinda_desilva@med.unc.edu.
authors.forename: Aravinda M
authors.initials: AM
authors.lastname: de Silva
mesh_terms: D056704:Adaptive Immunity; D000330:Aedes; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood* / Q000187:drug effects; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003715:Dengue / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D003716:Dengue Virus / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D007156:Immunologic Memory; D011485:Protein Binding; D000072417:Protein Domains; D014611:Vaccination; D014613:Vaccines, Attenuated / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014613:Vaccines, Attenuated; D014765:Viral Vaccines
keywords: Dengue virus serotype 4; antibody responses; epitope; human; infection; memory B cells; neutralization; vaccination