title: Human neutrophil flow chamber adhesion assay.
abstract: Neutrophil firm adhesion to endothelial cells plays a critical role in inflammation in both health and disease. The process of neutrophil firm adhesion involves many different adhesion molecules including members of the β2 integrin family and their counter-receptors of the ICAM family. Recently, naturally occurring genetic variants in both β2 integrins and ICAMs are reported to be associated with autoimmune disease. Thus, the quantitative adhesive capacity of neutrophils from individuals with varying allelic forms of these adhesion molecules is important to study in relation to mechanisms underlying development of autoimmunity. Adhesion studies in flow chamber systems can create an environment with fluid shear stress similar to that observed in the blood vessel environment in vivo. Here, we present a method using a flow chamber assay system to study the quantitative adhesive properties of human peripheral blood neutrophils to human umbilical vein endothelial cell (HUVEC) and to purified ligand substrates. With this method, the neutrophil adhesive capacities from donors with different allelic variants in adhesion receptors can be assessed and compared. This method can also be modified to assess adhesion of other primary cell types or cell lines.
authors.affiliation: Genetics and Genomic Sciences Graduate Program, University of Alabama at Birmingham.
authors.forename: Yebin
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Birmingham Veterans Affairs Medical Center; Department of Pathology, University of Alabama at Birmingham; Department of Biomedical Engineering, University of Alabama at Birmingham.
authors.forename: Dennis F
authors.initials: DF
authors.lastname: Kucik
authors.affiliation: Department of Medicine, University of Alabama at Birmingham.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Szalai
authors.affiliation: Department of Medicine, University of Alabama at Birmingham; jedberg@uab.edu.
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Edberg
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism
chemical_list: D015815:Cell Adhesion Molecules