Measurement of neutrophil adhesion under conditions mimicking blood flow.

Department of Veterinary Molecular Biology, Montana State University, Bozeman, MT, USA.
Methods in molecular biology (Clifton, N.J.) 02/2007; 412:239-56. DOI:10.1007/978-1-59745-467-4_16
Source: PubMed

ABSTRACT Neutrophil migration from blood into tissues is required for effective innate immune responses against infection. Adhesion of the neutrophil in blood to the vascular endothelium and eventual migration through the vessel wall and accumulation at the site of infection involves different classes of adhesion molecules. In vivo intravital microscopy studies show that different adhesion molecules mediate binding events under shear forces associated with blood flow vs binding events that take place under static conditions. To fully analyze the function of these adhesion molecules in vitro, assays must reflect the hemodynamic forces associated with blood flow. We outline two approaches used to study neutrophil adhesion under conditions that mimic blood flow.

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    ABSTRACT: The recruitment of leukocytes from circulation to sites of inflammation requires several families of adhesion molecules among which are selectins expressed on a variety of cells. In addition, they have also been shown to play key roles in the activation of cells in inflammation. To explore the collective role of E-, L-, and P- selectins in OVA-induced Th2 mediated response in acute asthma pathophysiology, ELP-/- mice were used and compared with age-matched wildtype (WT). Asthma phenotype was assessed by measuring pulmonary function, inflammation and OVA-specific serum IgE, which were completely abrogated in ELP-/- mice. Adoptive transfer of sensitized L selectin+CD4+ T cells into naïve ELP-/- mice which post-OVA challenge, developed asthma, suggesting that L-selectin may be critically involved in the onset of Th2 response in asthma. Tissue resident ELP-deficient cells were otherwise functionally competent as proved by normal proliferative response. Conclusions: Comparative studies between ELP-/- and WT mice uncovered functional roles of these three integrins in inflammatory response in allergic asthma. All three selectins seem to impede inflammatory migration while only L-selectin also possibly regulates activation of specific T cell subsets in lung and airways.
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