title: Label-free characterization of cell adhesion using reflectometric interference spectroscopy (RIfS).
abstract: Reflectometric interference spectroscopy (RIfS) is a label-free, time-resolved technique for detecting interactions of molecules immobilized on a surface with ligands in solution. Here we show that RIfS also permits the detection of the adhesion of tissue culture cells to a functionalized surface in a flow system. Interactions of T cells with other leukocytes or epithelial cells of blood vessels are crucial steps in the regulating immune response and inflammatory reactions. Jurkat T cell leukemia cells rapidly attached to a transducer functionalized with a monoclonal antibody directed against the T cell receptor (TCR)/CD3 complex, followed by activation-dependent cell spreading. RIfS curves were obtained for the Jurkat derivative JCaM 1.6 (which lacks the key signaling protein Lck), cells preincubated with cytochalasin D (an inhibitor of actin polymerization), and for surfaces functionalized with an antibody directed against the coreceptor CD28. These curves differed with respect to the maximum signal and the initial slope of the increase in optical thickness. The testing of chemical inhibitors, cell surface molecules and gene products relevant to a key event in T cell immunity illustrates the potential of label-free techniques for the analysis of activation-dependent cell-surface contacts.
authors.affiliation: Institute of Physical and Theoretical Chemistry, Eberhard-Karls-University of Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany. bernd.moehrle@ipc.uni-tuebingen.de
authors.forename: Bernd P
authors.initials: BP
authors.lastname: Möhrle
authors.forename: Karsten
authors.initials: K
authors.lastname: Köhler
authors.forename: Jan
authors.initials: J
authors.lastname: Jaehrling
authors.forename: Roland
authors.initials: R
authors.lastname: Brock
authors.forename: Günter
authors.initials: G
authors.lastname: Gauglitz
mesh_terms: D000906:Antibodies / Q000276:immunology; D002448:Cell Adhesion / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D015458:Leukemia, T-Cell / Q000235:genetics / Q000473:pathology*; D009154:Mutation / Q000235:genetics; D013057:Spectrum Analysis / Q000379:methods*
chemical_list: D000906:Antibodies