Authors: Bernd P Möhrle Karsten Köhler Jan Jaehrling Roland Brock Günter Gauglitz
Publish Date: 2005/12/01
Volume: 384, Issue: 2, Pages: 407-413
Abstract
Reflectometric interference spectroscopy RIfS is a labelfree timeresolved 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 activationdependent cell spreading RIfS curves were obtained for the Jurkat derivative JCaM 16 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 labelfree techniques for the analysis of activationdependent cellsurface contactsWe thank Ludger GrosseHovest and Gundram Jung for providing antibodies The JCaM 16 cell line was a kind gift from Claus Belka Roland Brock gratefully acknowledges financial support from the Volkswagen Foundation Nachwuchsgruppen an Universitäten Jan Jaehrling and Bernd Möhrle were participants of the research training group Graduiertenkolleg 8 “Quantitative Analysis and Characterisation of Pharmaceutically and Biochemically relevant Substances” and Karsten Köhler is a member of the Graduiertenkolleg 794 “Cellular Mechanisms of ImmuneAssociated Processes” both funded by the Deutsche Forschungsgemeinschaft DFG at the Eberhard Karls University of Tübingen
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