title: The nanoscale spatial organization of B-cell receptors on immunoglobulin M- and G-expressing human B-cells.
abstract: B-cell activation is initiated by the binding of antigen to the B-cell receptor (BCR). Here we used dSTORM superresolution imaging to characterize the nanoscale spatial organization of immunoglobulin M (IgM) and IgG BCRs on the surfaces of resting and antigen--activated human peripheral blood B-cells. We provide insights into both the fundamental process of antigen-driven BCR clustering and differences in the spatial organization of IgM and IgG BCRs that may contribute to the characteristic differences in the responses of naive and memory B-cells to antigen. We provide evidence that although both IgM and IgG BCRs reside in highly heterogeneous protein islands that vary in size and number of BCR single-molecule localizations, both resting and activated B-cells intrinsically maintain a high -frequency of single isolated BCR localizations, which likely represent BCR monomers. IgG BCRs are more clustered than IgM BCRs on resting cells and form larger protein islands after antigen activation. Small, dense BCR clusters likely formed via protein-protein interactions are present on the surface of resting cells, and antigen activation induces these to come together to form less dense, larger islands, a process likely governed, at least in part, by protein-lipid interactions.
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852.
authors.forename: Jinmin
authors.initials: J
authors.lastname: Lee
authors.affiliation: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.|Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Prabuddha
authors.initials: P
authors.lastname: Sengupta
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852.
authors.forename: Joseph
authors.initials: J
authors.lastname: Brzostowski
authors.affiliation: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.|Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lippincott-Schwartz
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852 spierce@nih.gov.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Pierce
mesh_terms: D000941:Antigens / Q000378:metabolism; D001402:B-Lymphocytes / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism / Q000502:physiology; D007075:Immunoglobulin M / Q000378:metabolism / Q000502:physiology; D008213:Lymphocyte Activation; D011947:Receptors, Antigen, B-Cell / Q000276:immunology* / Q000378:metabolism* / Q000648:ultrastructure; D015398:Signal Transduction / Q000276:immunology; D062211:Spatio-Temporal Analysis
chemical_list: D000941:Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011947:Receptors, Antigen, B-Cell