title: Nano-clustering of ligands on surrogate antigen presenting cells modulates T cell membrane adhesion and organization.
abstract: We investigate the adhesion and molecular organization of the plasma membrane of T lymphocytes interacting with a surrogate antigen presenting cell comprising glass supported ordered arrays of antibody (α-CD3) nano-dots dispersed in a non-adhesive matrix of polyethylene glycol (PEG). The local membrane adhesion and topography, as well as the distribution of the T cell receptors (TCRs) and the kinase ZAP-70, are influenced by dot-geometry, whereas the cell spreading area is determined by the overall average density of the ligands rather than specific characteristics of the dots. TCR clusters are recruited preferentially to the nano-dots and the TCR cluster size distribution has a weak dot-size dependence. On the patterns, the clusters are larger, more numerous, and more enriched in TCRs, as compared to the homogeneously distributed ligands at comparable concentrations. These observations support the idea that non-ligated TCRs residing in the non-adhered parts of the proximal membrane are able to diffuse and enrich the existing clusters at the ligand dots. However, long distance transport is impaired and cluster centralization in the form of a central supramolecular cluster (cSMAC) is not observed. Time-lapse imaging of early cell-surface contacts indicates that the ZAP-70 microclusters are directly recruited to the site of the antibody dots and this process is concomitant with membrane adhesion. These results together point to a complex interplay of adhesion, molecular organization and activation in response to spatially modulated stimulation.
authors.affiliation: Aix-Marseille Université, CNRS, CINaM-UMR 7325, Marseille, 13288, France. sengupta@cinam.univ-mrs.fr and Laboratoire Adhésion & Inflammation Aix-Marseille Université\Inserm U1067\CNRS-UMR7333, Marseille 13288, France. laurent.limozin@inserm.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Dillard
authors.affiliation: Aix-Marseille Université, CNRS, CINaM-UMR 7325, Marseille, 13288, France. sengupta@cinam.univ-mrs.fr.
authors.forename: Fuwei
authors.initials: F
authors.lastname: Pi
authors.affiliation: Laboratoire Adhésion & Inflammation Aix-Marseille Université\Inserm U1067\CNRS-UMR7333, Marseille 13288, France. laurent.limozin@inserm.fr.
authors.forename: Annemarie C
authors.initials: AC
authors.lastname: Lellouch
authors.affiliation: Laboratoire Adhésion & Inflammation Aix-Marseille Université\Inserm U1067\CNRS-UMR7333, Marseille 13288, France. laurent.limozin@inserm.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Limozin
authors.affiliation: Aix-Marseille Université, CNRS, CINaM-UMR 7325, Marseille, 13288, France. sengupta@cinam.univ-mrs.fr.
authors.forename: Kheya
authors.initials: K
authors.lastname: Sengupta
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D002448:Cell Adhesion / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D049329:Nanostructures; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D015398:Signal Transduction; D059010:Single-Cell Analysis; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000378:metabolism
chemical_list: D008024:Ligands; D011948:Receptors, Antigen, T-Cell; D049452:Green Fluorescent Proteins; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human