The Actin and Tetraspanin Networks Organize Receptor Nanoclusters to Regulate B Cell Receptor-Mediated Signaling

• Published in: Immunity (Cambridge, Mass.) Mass.), 2013-03, Vol.38 (3), p.461-474
• Main Authors: Mattila, Pieta K., Feest, Christoph, Depoil, David, Treanor, Bebhinn, Montaner, Beatriz, Otipoby, Kevin L., Carter, Robert, Justement, Louis B., Bruckbauer, Andreas, Batista, Facundo D.
• Format: Article
• Language: English
• Subjects: Actins - immunology; Actins - metabolism; Animals; Antigens; Antigens, CD19 - genetics; Antigens, CD19 - immunology; Antigens, CD19 - metabolism; B-Lymphocytes - immunology; B-Lymphocytes - metabolism; Cell Membrane - immunology; Cell Membrane - metabolism; Cytoskeleton; Cytoskeleton - immunology; Cytoskeleton - metabolism; Flow Cytometry; Immune system; Immunoblotting; Immunoglobulin D - immunology; Immunoglobulin D - metabolism; Immunoglobulin M - immunology; Immunoglobulin M - metabolism; Kinases; Medical research; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Microscopy, Fluorescence; Models, Immunological; Nanostructures; Protein Binding - immunology; Receptors, Antigen, B-Cell - immunology; Receptors, Antigen, B-Cell - metabolism; Rodents; Signal Transduction - immunology; Tetraspanin 28 - genetics; Tetraspanin 28 - immunology; Tetraspanin 28 - metabolism
• ISSN: 1074-7613; 1097-4180
• DOI: 10.1016/j.immuni.2012.11.019

A key role is emerging for the cytoskeleton in coordinating receptor signaling, although the underlying molecular requirements remain unclear. Here we show that cytoskeleton disruption triggered signaling requiring not only the B cell receptor (BCR), but also the coreceptor CD19 and tetraspanin CD81, thus providing a mechanism for signal amplification upon surface-bound antigen stimulation. By using superresolution microscopy, we demonstrated that endogenous IgM, IgD, and CD19 exhibited distinct nanoscale organization within the plasma membrane of primary B cells. Upon stimulation, we detect a local convergence of receptors, although their global organization was not dramatically altered. Thus, we postulate that cytoskeleton reorganization releases BCR nanoclusters, which can interact with CD19 held in place by the tetraspanin network. These results not only suggest that receptor compartmentalization regulates antigen-induced activation but also imply a potential role for CD19 in mediating ligand-independent “tonic” BCR signaling necessary for B cell survival. [Display omitted] ► In resting cells, the BCR and the coreceptor CD19 are organized in nanoclusters ► Differential compartmentalization of the BCR and CD19 is important for BCR signaling ► Disruption of the actin cytoskeleton triggers signaling through the BCR and CD19 ► This antigen-independent signaling requires CD19 organized by the tetraspanin network