title: Vaccine-induced plasmablast responses in rhesus macaques: phenotypic characterization and a source for generating antigen-specific monoclonal antibodies.
abstract: Over 100 broadly neutralizing antibodies have been isolated from a minority of HIV infected patients, but the steps leading to the selection of plasma cells producing such antibodies remain incompletely understood, hampering the development of vaccines able to elicit them. Rhesus macaques have become a preferred animal model system used to study SIV/HIV, for the characterization and development of novel therapeutics and vaccines as well as to understand pathogenesis. However, most of our knowledge about the dynamics of antibody responses is limited to the analysis of serum antibodies or monoclonal antibodies generated from memory B cells. In a vaccine setting, relatively little is known about the early cellular responses that elicit long-lived plasma cells and memory B cells and the tools to dissect plasmablast responses are not available in macaques. In the current study, we show that the majority (>80%) of the vaccine-induced plasmablast response are antigen-specific by functional ELISPOT assays. While plasmablasts are easily defined and isolated in humans, those same phenotypic markers have not been useful for identifying macaque plasmablasts. Here we describe an approach that allows for the isolation and single cell sorting of vaccine-induced plasmablasts. Finally, we show that isolated plasmablasts can be used to efficiently recover antigen-specific monoclonal antibodies through single cell expression cloning. This will allow detailed studies of the early plasmablast responses in rhesus macaques, enabling the characterization of both their repertoire breadth as well as the epitope specificity and functional qualities of the antibodies they produce, not only in the context of SIV/HIV vaccines but for many other pathogens/vaccines as well.
authors.affiliation: Department of Pediatrics and Department of Hematology and Oncology, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA; Division of Microbiology and Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA, 30329, USA.
authors.forename: Eduardo L V
authors.initials: EL
authors.lastname: Silveira
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA.
authors.forename: Sudhir P
authors.initials: SP
authors.lastname: Kasturi
authors.affiliation: Department of Pediatrics and Department of Hematology and Oncology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Yevgeniy
authors.initials: Y
authors.lastname: Kovalenkov
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA.
authors.forename: Ata Ur
authors.initials: AU
authors.lastname: Rasheed
authors.affiliation: Department of Pediatrics and Department of Hematology and Oncology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Patryce
authors.initials: P
authors.lastname: Yeiser
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA; Division of Microbiology and Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA, 30329, USA.
authors.forename: Zarpheen S
authors.initials: ZS
authors.lastname: Jinnah
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA; Division of Microbiology and Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA, 30329, USA.
authors.forename: Traci H
authors.initials: TH
authors.lastname: Legere
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA; Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.
authors.forename: Bali
authors.initials: B
authors.lastname: Pulendran
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA; Division of Microbiology and Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA, 30329, USA; Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.
authors.forename: Francois
authors.initials: F
authors.lastname: Villinger
authors.affiliation: Department of Pediatrics and Department of Hematology and Oncology, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Vaccine Center, Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329, USA. Electronic address: jwramme@emory.edu.
authors.forename: Jens
authors.initials: J
authors.lastname: Wrammert
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology; D000941:Antigens / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D006678:HIV / Q000276:immunology; D015658:HIV Infections / Q000276:immunology; D007156:Immunologic Memory / Q000276:immunology; D008253:Macaca mulatta / Q000276:immunology* / Q000821:virology; D023421:Models, Animal; D010950:Plasma Cells / Q000276:immunology*; D018449:SAIDS Vaccines / Q000276:immunology; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology; D015302:Simian Immunodeficiency Virus / Q000276:immunology; D014612:Vaccines / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000941:Antigens; D018449:SAIDS Vaccines; D014612:Vaccines
keywords: Macaque plasmablasts; Monoclonal antibodies; Phenotype; Sorting