title: The SW(HEL) system for high-resolution analysis of in vivo antigen-specific T-dependent B cell responses.
abstract: T cell-dependent B cell responses generate optimal antibodies to combat foreign antigens. Naïve B cells responding to antigen undergo a complex series of differentiation events and cell fate decisions to provide long-lived memory B cells and plasma cells. Historically, B cell biologists have been challenged by the task of investigating rare antigen-specific B cells in an in vivo setting such that their interactions with antigen, regulation and migration may be accurately tracked. We have developed the SW(HEL) experimental system capable of accurately monitoring B cells that interact with a protein antigen and then subsequently undergo isotype switching, somatic hypermutation, and affinity maturation within germinal centers (GC) to generate high-affinity antibodies. Here we provide a comprehensive description of the procedures involved in establishing and using the SW(HEL) system to assess B cell responses to a foreign antigen. This system can provide a valuable measure of the functional capabilities of T follicular helper cells, whose role is ultimately to support and shape long-term humoral immunity.
authors.affiliation: Immunology Division, Garvan Institute of Medical Research, 384 Victoria St., Darlinghurst, NSW, 2010, Australia, r.brink@garvan.org.au.
authors.forename: R
authors.initials: R
authors.lastname: Brink
authors.forename: D
authors.initials: D
authors.lastname: Paus
authors.forename: K
authors.initials: K
authors.lastname: Bourne
authors.forename: J R
authors.initials: JR
authors.lastname: Hermes
authors.forename: S
authors.initials: S
authors.lastname: Gardam
authors.forename: T G
authors.initials: TG
authors.lastname: Phan
authors.forename: T D
authors.initials: TD
authors.lastname: Chan
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D000906:Antibodies / Q000097:blood; D000915:Antibody Affinity / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D004912:Erythrocytes / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D060005:Genotyping Techniques; D007158:Immunologic Techniques / Q000379:methods*; D016131:Lymphocyte Subsets / Q000276:immunology; D008822:Mice, Transgenic; D009113:Muramidase / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D012756:Sheep; D027041:Somatic Hypermutation, Immunoglobulin; D013154:Spleen / Q000166:cytology; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000906:Antibodies; D000939:Epitopes; D011994:Recombinant Proteins; D009113:Muramidase