title: Dynamics of Dengue Virus (DENV)-Specific B Cells in the Response to DENV Serotype 1 Infections, Using Flow Cytometry With Labeled Virions.
abstract: BACKGROUND
  The development of reagents to identify and characterize antigen-specific B cells has been challenging.
  
  
  METHODS
  We recently developed Alexa Fluor-labeled dengue viruses (AF DENVs) to characterize antigen-specific B cells in the peripheral blood of DENV-immune individuals.
  
  
  RESULTS
  In this study, we used AF DENV serotype 1 (AF DENV-1) together with AF DENV-2 on peripheral blood mononuclear cells (PBMCs) from children in Thailand with acute primary or secondary DENV-1 infections to analyze the phenotypes of antigen-specific B cells that reflected their exposure or clinical diagnosis. DENV serotype-specific and cross-reactive B cells were identified in PBMCs from all subjects. Frequencies of AF DENV(+) class-switched memory B cells (IgD(-)CD27(+) CD19(+) cells) reached up to 8% during acute infection and early convalescence. AF DENV-labeled B cells expressed high levels of CD27 and CD38 during acute infection, characteristic of plasmablasts, and transitioned into memory B cells (CD38(-)CD27(+)) at the early convalescent time point. There was higher activation of memory B cells early during acute secondary infection, suggesting reactivation from a previous DENV infection.
  
  
  CONCLUSIONS
  AF DENVs reveal changes in the phenotype of DENV serotype-specific and cross-reactive B cells during and after natural DENV infection and could be useful in analysis of the response to DENV vaccination.
authors.affiliation: Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester Institute for Immunology and Informatics, University of Rhode Island, Providence.
authors.forename: Marcia
authors.initials: M
authors.lastname: Woda
authors.affiliation: Walter Reed Army Institute of Research, Silver Spring, Maryland.
authors.forename: Heather
authors.initials: H
authors.lastname: Friberg
authors.affiliation: Walter Reed Army Institute of Research, Silver Spring, Maryland.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Currier
authors.affiliation: Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
authors.forename: Anon
authors.initials: A
authors.lastname: Srikiatkhachorn
authors.affiliation: Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
authors.forename: Louis R
authors.initials: LR
authors.lastname: Macareo
authors.affiliation: Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester.
authors.forename: Sharone
authors.initials: S
authors.lastname: Green
authors.affiliation: Walter Reed Army Institute of Research, Silver Spring, Maryland.
authors.forename: Richard G
authors.initials: RG
authors.lastname: Jarman
authors.affiliation: Institute for Immunology and Informatics, University of Rhode Island, Providence.
authors.forename: Alan L
authors.initials: AL
authors.lastname: Rothman
authors.affiliation: Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester Institute for Immunology and Informatics, University of Rhode Island, Providence.
authors.forename: Anuja
authors.initials: A
authors.lastname: Mathew
mesh_terms: D000293:Adolescent; D001402:B-Lymphocytes / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D003715:Dengue / Q000276:immunology* / Q000821:virology*; D003716:Dengue Virus / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007223:Infant; D008297:Male; D065288:Serogroup; D013194:Staining and Labeling / Q000379:methods; D013785:Thailand; D014771:Virion / Q000276:immunology
keywords: Alexa Fluor; B cell; Dengue; antigen; flow cytometry