Vaccine-induced plasmablast responses in rhesus macaques: Phenotypic characterization and a source for generating antigen-specific monoclonal antibodies

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 becom...

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Bibliographic Details
Published in:	Journal of immunological methods Vol. 416; pp. 69 - 83
Main Authors:	Silveira, Eduardo L.V., Kasturi, Sudhir P., Kovalenkov, Yevgeniy, Rasheed, Ata Ur, Yeiser, Patryce, Jinnah, Zarpheen S., Legere, Traci H., Pulendran, Bali, Villinger, Francois, Wrammert, Jens
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01-01-2015
Subjects:	AIDS Vaccines - immunology Animals Antibodies, Monoclonal - immunology Antibodies, Neutralizing - immunology Antigens - immunology B-Lymphocytes - immunology Cell Separation - methods Enzyme-Linked Immunospot Assay - methods HIV - immunology HIV Infections - immunology Immunologic Memory - immunology Macaca mulatta - immunology Macaca mulatta - virology Macaque plasmablasts Models, Animal Monoclonal antibodies Phenotype Plasma Cells - immunology SAIDS Vaccines - immunology Simian Acquired Immunodeficiency Syndrome - immunology Simian Immunodeficiency Virus - immunology Sorting Vaccines - immunology Monoclonal antibodies Macaque plasmablasts Phenotype Sorting
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Summary:	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. •Potent SIV gp140-specific plasmablast responses after vaccination in rhesus macaques•The majority of the plasmablasts were antigen-specific at the peak of the response.•After booster vaccination, macaque plasmablasts appear earlier than in humans.•Phenotypic/functional analyses of vaccine-induced plasmablasts enabled their isolation.•Single cell plasmablasts as a source of antigen-specific monoclonal antibodies
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0022-1759 1872-7905
DOI:	10.1016/j.jim.2014.11.003