Superresolution imaging reveals nanometer- and micrometer-scale spatial distributions of T-cell receptors in lymph nodes.
T cells become activated when T-cell receptors (TCRs) recognize agonist peptides bound to major histocompatibility complex molecules on antigen-presenting cells. T-cell activation critically relies on the spatiotemporal arrangements of TCRs on the plasma membrane. However, the molecular organizations of TCRs on lymph node-resident T cells have not yet been determined, owing to the diffraction limit of light. Here we visualized nanometer- and micrometer-scale TCR distributions in lymph nodes by light sheet direct stochastic optical reconstruction microscopy (dSTORM) and structured illumination microscopy (SIM). This dSTORM and SIM approach provides the first evidence, to our knowledge, of multiscale reorganization of TCRs during in vivo immune responses. We observed nanometer-scale plasma membrane domains, known as protein islands, on naïve T cells. These protein islands were enriched within micrometer-sized surface areas that we call territories. In vivo T-cell activation caused the TCR territories to contract, leading to the coalescence of protein islands and formation of stable TCR microclusters.
Nomis Center for Immunobiology and Microbial Pathogenesis, Salk Institute for Biological Studies, La Jolla, CA 92037; Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037.
Ying S
YS
Hu
Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037 blillemeier@salk.edu hucang@salk.edu.
Hu
H
Cang
Nomis Center for Immunobiology and Microbial Pathogenesis, Salk Institute for Biological Studies, La Jolla, CA 92037; Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037 blillemeier@salk.edu hucang@salk.edu.
Björn F
BF
Lillemeier
D000818:Animals; D045304:Cytochromes c / Q000276:immunology; D003952:Diagnostic Imaging / Q000379:methods; D019476:Insect Proteins / Q000276:immunology; D008198:Lymph Nodes / Q000000981:diagnostic imaging* / Q000276:immunology*; D008822:Mice, Transgenic; D036103:Nanotechnology / Q000379:methods; D010455:Peptides / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*
D019476:Insect Proteins; D010455:Peptides; D011948:Receptors, Antigen, T-Cell; D045304:Cytochromes c
T-cell activation; T-cell receptor; plasma membrane; protein cluster; superresolution