title: Optical and fluorescence detection of neutrophil integrin activation.
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.
authors.affiliation: Department of Biomedical Engineering, University of California, Davis, CA, USA.
authors.forename: Ulrich Y
authors.initials: UY
authors.lastname: Schaff
authors.forename: Melissa R
authors.initials: MR
authors.lastname: Sarantos
authors.forename: Harold
authors.initials: H
authors.lastname: Ting
authors.forename: Scott I
authors.initials: SI
authors.lastname: Simon
mesh_terms: D000954:Antigens, Surface / Q000032:analysis; D000939:Epitopes; D005453:Fluorescence; D016023:Integrins / Q000378:metabolism*; D008954:Models, Biological; D018375:Neutrophil Activation / Q000502:physiology*; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D055095:Optics and Photonics
chemical_list: D000954:Antigens, Surface; D000939:Epitopes; D016023:Integrins