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Detection of Bidirectional Signaling During Integrin Activation and Neutrophil Adhesion

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Neutrophil Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1124))

Abstract

Neutrophil arrest and migration on inflamed endothelium is dependent upon a conformational shift in CD11a/CD18 (LFA-1) from a low to high affinity and clustered state which determines the strength and lifetime of bond formation with intracellular adhesion molecule 1 (ICAM-1). Cytoskeletal adaptor proteins kindlin-3 and talin-1 anchor clustered LFA-1 to the cytoskeleton and support the transition from neutrophil rolling to arrest. We employ microfluidic flow channels and total internal reflection fluorescence microscopy to evaluate the spatiotemporal regulation of LFA-1 affinity and bond formation that facilitate the transition from neutrophil rolling to arrest. Methodology is presented to correlate the relationship between integrin conformation, bond formation with ICAM-1, and cytoskeletal engagement and adhesion strengthening necessary to achieve a migratory phenotype.

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Acknowledgements

This work was supported by National Institutes of Health (NIH) grant AI472294 to S.I.S.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, Davis, CA, USA

    Stuart M. Altman, Neha Dixit & Scott I. Simon

Authors
  1. Stuart M. Altman
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  2. Neha Dixit
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  3. Scott I. Simon
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Editor information

Editors and Affiliations

  1. Montana State University Dept. Veterinary Molecular Biology, Bozeman, Montana, USA

    Mark T. Quinn

  2. Lab. Human Bacterial Pathogenesis, National Institutes of Health Nat. Inst. Allergy & Infectious Diseases, Hamilton, Montana, USA

    Frank R. DeLeo

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© 2014 Springer Science+Business Media, LLC

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Altman, S.M., Dixit, N., Simon, S.I. (2014). Detection of Bidirectional Signaling During Integrin Activation and Neutrophil Adhesion. In: Quinn, M., DeLeo, F. (eds) Neutrophil Methods and Protocols. Methods in Molecular Biology, vol 1124. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-845-4_15

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  • DOI: https://doi.org/10.1007/978-1-62703-845-4_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-844-7

  • Online ISBN: 978-1-62703-845-4

  • eBook Packages: Springer Protocols

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