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Label-free characterization of cell adhesion using reflectometric interference spectroscopy (RIfS)

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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.

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Acknowledgements

We thank Ludger Grosse-Hovest and Gundram Jung for providing antibodies. The JCaM 1.6 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 Immune-Associated Processes”, both funded by the Deutsche Forschungsgemeinschaft (DFG) at the Eberhard Karls University of Tübingen.

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Authors and Affiliations

  1. Institute of Physical and Theoretical Chemistry, Eberhard-Karls-University of Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany

    Bernd P. Möhrle, Jan Jaehrling & Günter Gauglitz

  2. Department of Molecular Biology, Institute for Cell Biology, Eberhard-Karls-University of Tübingen, Auf der Morgenstelle 15, 72076, Tübingen, Germany

    Karsten Köhler & Roland Brock

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  1. Bernd P. Möhrle
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  2. Karsten Köhler
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  3. Jan Jaehrling
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  4. Roland Brock
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  5. Günter Gauglitz
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Corresponding author

Correspondence to Bernd P. Möhrle.

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The first two authors contributed equally to this paper

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Möhrle, B.P., Köhler, K., Jaehrling, J. et al. Label-free characterization of cell adhesion using reflectometric interference spectroscopy (RIfS). Anal Bioanal Chem 384, 407–413 (2006). https://doi.org/10.1007/s00216-005-0202-7

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  • DOI: https://doi.org/10.1007/s00216-005-0202-7

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