Mechanical Shedding of L-selectin from the Neutrophil Surface during Rolling on Sialyl Lewis x under Flow

Lee, Dong Hoon; Schultz, Joanne; Knauf, Philip A.; King, Michael R.

doi:10.1074/jbc.m609994200

Cited by 73 publications

(94 citation statements)

References 46 publications

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“…Neutrophil movement on the HUVEC monolayer was tracked with Metamorph software (Molecular Devices). Cells were regarded as rolling when they met the following two conditions: 1) when they moved on endothelial cells for at least 2 s while remaining in the field of view, and 2) when their average velocity was ,50% of that calculated for noninteracting neutrophils (29) during the first 9 min of recording. Cells that had observable phase-bright and phase-dark periods at any time during the 6 min after the rolling period were considered transmigrated.…”

Section: Flow Chamber Experimentsmentioning

confidence: 99%

Prevention of Neutrophil Extravasation by α2-Adrenoceptor–Mediated Endothelial Stabilization

Herrera-García

Domínguez-Luis

Arce-Franco

et al. 2014

The Journal of Immunology

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Adrenergic receptors are expressed on the surface of inflammation-mediating cells, but their potential role in the regulation of the inflammatory response is still poorly understood. The objectives of this work were to study the effects of α2-adrenergic agonists on the inflammatory response in vivo and to determine their mechanism of action. In two mouse models of inflammation, zymosan air pouch and thioglycolate-induced peritonitis models, the i.m. treatment with xylazine or UK14304, two α2-adrenergic agonists, reduced neutrophil migration by 60%. The α2-adrenergic antagonist RX821002 abrogated this effect. In flow cytometry experiments, the basal surface expression of L-selectin and CD11b was modified neither in murine nor in human neutrophils upon α2-agonist treatment. Similar experiments in HUVEC showed that UK14304 prevented the activation-dependent upregulation of ICAM-1. In contrast, UK14304 augmented electrical resistance and reduced macromolecular transport through a confluent HUVEC monolayer. In flow chamber experiments, under postcapillary venule-like flow conditions, the pretreatment of HUVECs, but not neutrophils, with α2-agonists decreased transendothelial migration, without affecting neutrophil rolling. Interestingly, α2-agonists prevented the TNF-α–mediated decrease in expression of the adherens junctional molecules, VE-cadherin, β-catenin, and plakoglobin, and reduced the ICAM-1–mediated phosphorylation of VE-cadherin by immunofluorescence and confocal analysis and Western blot analysis, respectively. These findings indicate that α2-adrenoceptors trigger signals that protect the integrity of endothelial adherens junctions during the inflammatory response, thus pointing at the vascular endothelium as a therapeutic target for the management of inflammatory processes in humans.

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Section: Flow Chamber Experimentsmentioning

confidence: 99%

Prevention of Neutrophil Extravasation by α2-Adrenoceptor–Mediated Endothelial Stabilization

Herrera-García

Domínguez-Luis

Arce-Franco

et al. 2014

The Journal of Immunology

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“…On the contrary, RANTES-recruited neutrophils present a low side scatter and high Gr1 expression, and they are L-selectin-positive and CCR3-and CCR5-negative. Molecules of the selectin family such as L-selectin are constitutively expressed on leukocytes and shed from the cell surface upon activation (Lee et al, 2007). Therefore, it is possible that neutrophils transiently recruited by RANTES in the absence of additional inflammatory signals do not become activated in the absence of local inflammatory stimuli.…”

Section: Discussionmentioning

confidence: 99%

Phosphoinositide 3-Kinase γ Inhibition Plays a Crucial Role in Early Steps of Inflammation by Blocking Neutrophil Recruitment

Ferrandi

Ardissone²,

Ferro³

et al. 2007

J Pharmacol Exp Ther

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Leukocyte trafficking to inflammatory sites is a gradual process, which is dominated in its early phases by chemokine-and cytokine-mediated neutrophil recruitment. The chemokine regulated on activation normal T cell expressed and secreted (RANTES) has been shown to be highly expressed in the joints of patient with rheumatoid arthritis and to promote leukocyte trafficking into the synovial tissue. In this study, we investigated the effect of RANTES in a murine model of peritoneal chemotaxis, and we found that RANTES dose-dependently induces neutrophil recruitment. Then, through morphological and histological analyses, we observed that activated neutrophils represent the major infiltrating population in response to RANTES chemotactic stimulus. Furthermore, we demonstrated that oral administration of either nonisoform-specific phosphoinositide 3-kinase (PI3K) inhibitor LY294002 (morpholin-4-yl-8-phenylchromen-4-one) or selective PI3K␥ inhibitor AS041164 (5-benzo[1,3]dioxol-5-ylmethylene-thiazolidine-2,4-dione) blocks RANTES-induced chemotaxis and reduces the level of AKT phosphorylation. Because the two compounds showed a similar pharmacokinetic profile in terms of bioavailability and half-life after oral route administration, the selective inhibition of the PI3K␥-isoform pathway through AS041164 was three times more potent in reducing neutrophil recruitment. Finally, to confirm the blockade of neutrophil infiltration that occurs in the early phase of the inflammatory response, AS041164 was also tested in a model of carrageenan-induced paw edema in rats. Therefore, the PI3K␥ pathway plays an important role in controlling neutrophil chemotaxis during early steps of inflammation.During inflammation, neutrophils are rapidly recruited at sites of acute infection and dominate the initial influx of leukocytes (Issekutz and Movat, 1980). Later, during the progression of inflammation, monocytes and macrophages replace neutrophils, suggesting a bimodal recruitment pattern involving a switch from neutrophils to monocytes (Doherty et al., 1988;Henderson et al., 2003). The first step in approaching a site of insult requires neutrophils to transmigrate across endothelial barriers, a process that depends on chemokines (Yoshie et al., 2001). In response to a chemotactic gradient, CXC and CC chemokines activate leukocytes by binding to seven transmembrane receptors coupled to G proteins (Proudfoot, 2002) linked to heterotrimeric G protein complexes (Proudfoot, 2002). Upon stimulation, the G protein complex dissociates and subsequently recruits various signaling components, such as nucleotide exchange factors, phospholipid lipases, and lipid kinase phosphoinositide-3ЈOH-kinase isoforms, such as phosphoinositide 3-kinase (PI3K) (Akasaki et al., 1999). Neutrophils have been shown to express different chemokine receptors, including CXCR2 and CCR1 (Lee et al., 1995;Zhang et al., 1999). Furthermore, the blockade of the chemokine receptor CXCR2 or its ligands IL-8 and macrophage inflammatory protein (MIP)-2, or alternatively of t...

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“…on May 10, 2018. by guest www.bloodjournal.org From trafficking potential of neutrophils and T cells, respectively. 23,31,32 To test whether L-selectin shedding could account for the observed rolling defect, cell lines were challenged with increasing amounts of PMA, a potent artificial stimulator of L-selectin shedding. No differences in activation-induced shedding of L-selectin were seen ( Figure 6A).…”

Section: Wasp I294t Affects Lymphocyte Adhesion Under Flow 5359mentioning

confidence: 99%

“…Sialyl Lewis-X (sLe x ; GlycoTech) was prepared as previously described 23 (see supplemental Methods for more information). Excess sLe x was removed by aspiration and gentle washing with PBS, after which slides were blocked in PBS/1% bovine serum albumin (BSA) for 2 hours at room temperature.…”

Section: Parallel Plate Flow Chamber Assaymentioning

confidence: 99%

A congenital activating mutant of WASp causes altered plasma membrane topography and adhesion under flow in lymphocytes

Burns

Killock

Moulding

et al. 2010

Blood

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Leukocytes rely on dynamic actindependent changes in cell shape to pass through blood vessels, which is fundamental to immune surveillance. WiskottAldrich Syndrome protein (WASp) is a hematopoietic cell-restricted cytoskeletal regulator important for modulating cell shape through Arp2/3-mediated actin polymerization. A recently identified WASp I294T mutation was shown to render WASp constitutively active in vivo, causing increased filamentous (F)-actin polymerization, high podosome turnover in macrophages, and myelodysplasia. The aim of this study was to determine the effect of WASp I294T expression in lymphocytes. Here, we report that lymphocytes isolated from a patient with WASp I294T , and in a cellular model of WASp I294T IntroductionThe Wiskott-Aldrich Syndrome protein (WASp) is a key cytoskeletal regulator in hematopoietic cells. 1 Through its multidomain structure, WASp integrates inputs from disparate signaling pathways to initiate and regulate Arp2/3-mediated actin polymerization. This is important for the normal formation of various actin-rich structures including the T-cell immune synapse, phagocytic cups, and specialized adhesion structures called podosomes in myeloid cells. [2][3][4] The majority of human disease-causing mutations in WASp are hypomorphic or null loss of function mutations, which result in Wiskott-Aldrich Syndrome (WAS). 5 Patients with WAS classically suffer from thrombocytopenia, eczema, and a broad immunodeficiency, reflecting the functional deficiency of almost all hematopoietic lineages.Recently, 3 novel human mutations (L270P, S272P, and I294T) leading to constitutive WASp activation have been reported to cause a distinct disease, X-linked neutropoenia (XLN). 6,7 All 3 are positioned in the GTPase binding domain (GBD; residues 230-288) of WASp and disrupt its autoinhibitory interaction with the Arp2/3 binding, C-terminal verprolin homology, central, acidic (VCA) domain. The result is enhanced and dysregulated WASp activity characterized by an increase in cellular filamentous (F)-actin and abnormalities of actin cytoskeletal structure and dynamics. A large kindred bearing the rare I294T mutation was recently reported, informing our understanding of the clinical spectrum of WASp I294T disease, which appears variable but is usually associated with neutropoenia and recurrent infections. 7,8 Although fatal infections have been described, infectious complications are surprisingly mild and not correlated with the degree of neutropoenia. 8 Other immunopathology is also seen, including low levels of CD4 and CD8 T cells, natural killer (NK) cell and B-cell lymphopenia, and reduced levels of IgA. To date, specific cellular effects have only been examined in myeloid cells, where defects of myelopoiesis and podosome formation have been reported. 7,9 Our aim in this report was to determine whether WASp I294T also attenuates aspects of lymphocyte function.The lymphocyte surface is dominated by microvilli: prominent actin-rich, fingerlike membrane projections involved in mediating cell-c...

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Mechanical Shedding of L-selectin from the Neutrophil Surface during Rolling on Sialyl Lewis x under Flow

Cited by 73 publications

References 46 publications

Prevention of Neutrophil Extravasation by α2-Adrenoceptor–Mediated Endothelial Stabilization

Prevention of Neutrophil Extravasation by α2-Adrenoceptor–Mediated Endothelial Stabilization

Phosphoinositide 3-Kinase γ Inhibition Plays a Crucial Role in Early Steps of Inflammation by Blocking Neutrophil Recruitment

A congenital activating mutant of WASp causes altered plasma membrane topography and adhesion under flow in lymphocytes

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