title: Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function.
abstract: B cells reactive with a specific antigen usually occur at a frequency of <0.05% of lymphocytes. For decades researchers have sought methods to isolate and enrich these rare cells for studies of their phenotype and biology. Approaches are inevitably based on the principle that B cells recognize native antigen by virtue of cell surface receptors that are representative in specificity of antibodies that will eventually be secreted by their differentiated daughters. Perhaps the most obvious approach to the problem involves use of fluorochrome-conjugated antigens in conjunction with fluorescence-activated cell sorting (FACS). However, the utility of these methods is limited by cell frequency and the achievable rate of analysis and isolation by electronic sorting. A novel method to enrich rare antigen-specific B cells using magnetic nanoparticles that results in high yield enrichment of antigen-reactive B cells from large starting cell populations is described. This method enables improved monitoring of the phenotype and biology of antigen reactive cells before and following in vivo antigen encounter, such as after immunization or during development of autoimmunity.
authors.affiliation: Department of Immunology and Microbiology, University of Colorado School of Medicine; Department of Microbiology, Immunology, and Pathology, Colorado State University.
authors.forename: Mia J
authors.initials: MJ
authors.lastname: Smith
authors.affiliation: Department of Immunology and Microbiology, University of Colorado School of Medicine.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Packard
authors.affiliation: Department of Immunology and Microbiology, University of Colorado School of Medicine.
authors.forename: Shannon K
authors.initials: SK
authors.lastname: O'Neill
authors.affiliation: Department of Immunology and Microbiology, University of Colorado School of Medicine.
authors.forename: Rochelle M
authors.initials: RM
authors.lastname: Hinman
authors.affiliation: Barbara Davis Center for Childhood Diabetes, University of Colorado School of Medicine.
authors.forename: Marynette
authors.initials: M
authors.lastname: Rihanek
authors.affiliation: Barbara Davis Center for Childhood Diabetes, University of Colorado School of Medicine.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Gottlieb
authors.affiliation: Department of Immunology and Microbiology, University of Colorado School of Medicine; Department of Biomedical Research, National Jewish Health; john.cambier@ucdenver.edu.
authors.forename: John C
authors.initials: JC
authors.lastname: Cambier
mesh_terms: D000921:Antibody-Producing Cells / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D060526:Magnetic Fields; D053758:Nanoparticles; D010641:Phenotype