Optical and fluorescence detection of neutrophil integrin activation.

Ulrich Y. Schaff, Melissa R. Sarantos, Harold Ting, Scott I. Simon

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Neutrophils are among the first cells to respond to acute inflammation through a multistep process initiated by selectin mediated rolling, which transitions to an integrin/intercellular adhesion molecule-dependent arrest and transmigration across endothelium. A conformational shift in the CD11/CD18 adhesion receptor on neutrophils is a critical determinant of the efficiency of recruitment on inflamed endothelium. For instance, beta2-integrin expression level is upregulated up to 10-fold by fusion of cytoplasmic granule pools of CD11b/CD18 (Mac-1). Furthermore, a rapid increase in affinity and membrane clustering of CD11a/CD18 (LFA-1) is necessary for efficient deceleration and arrest in shear flow. We present methods here to quantify the changes in receptor expression and affinity that support neutrophil adhesive phenotypes. Techniques involving real-time fluorescence flow cytometry and parallel plate rheometry coupled with light microscopy are presented.

Original languageEnglish (US)
Pages (from-to)203-210
Number of pages8
JournalMethods in molecular biology (Clifton, N.J.)
Volume412
StatePublished - 2007

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Neutrophil Activation
Integrins
Neutrophils
Fluorescence
Endothelium
CD18 Antigens
Cytoplasmic Granules
Selectins
Lymphocyte Function-Associated Antigen-1
Deceleration
Cell Adhesion Molecules
Adhesives
Cluster Analysis
Microscopy
Flow Cytometry
Inflammation
Phenotype
Light
Membranes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Schaff, U. Y., Sarantos, M. R., Ting, H., & Simon, S. I. (2007). Optical and fluorescence detection of neutrophil integrin activation. Methods in molecular biology (Clifton, N.J.), 412, 203-210.

Optical and fluorescence detection of neutrophil integrin activation. / Schaff, Ulrich Y.; Sarantos, Melissa R.; Ting, Harold; Simon, Scott I.

In: Methods in molecular biology (Clifton, N.J.), Vol. 412, 2007, p. 203-210.

Research output: Contribution to journalArticle

Schaff, UY, Sarantos, MR, Ting, H & Simon, SI 2007, 'Optical and fluorescence detection of neutrophil integrin activation.', Methods in molecular biology (Clifton, N.J.), vol. 412, pp. 203-210.
Schaff, Ulrich Y. ; Sarantos, Melissa R. ; Ting, Harold ; Simon, Scott I. / Optical and fluorescence detection of neutrophil integrin activation. In: Methods in molecular biology (Clifton, N.J.). 2007 ; Vol. 412. pp. 203-210.
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