CD157 is an important mediator of neutrophil adhesion and migration

Blood ◽  
2004 ◽  
Vol 104 (13) ◽  
pp. 4269-4278 ◽  
Author(s):  
Ada Funaro ◽  
Erika Ortolan ◽  
Bruna Ferranti ◽  
Lucia Gargiulo ◽  
Rosario Notaro ◽  
...  
Keyword(s):  
Cell Shape ◽  
Adenosine Diphosphate ◽  
Enzymatic Activities ◽  
Matrix Proteins ◽  
Β2 Integrin ◽  
Important Mediator ◽  
Adp Ribosyl Cyclase ◽  
And Migration ◽  
Cyclic Adenosine Diphosphate

Abstract CD157, a glycosylphosphatidylinositol (GPI)–anchored protein encoded by a member of the CD38 NADase/ADP-ribosyl cyclase gene family, is expressed on the surface of most human circulating neutrophils. This work demonstrates that CD157 is a receptor that induces reorganization of the cytoskeleton and significant changes in cell shape, and that signals mediated by CD157 act through modulation of cytosolic Ca2+ concentration. These signals are independent of the products of CD157's enzymatic activities (ie, cyclic adenosine diphosphate [ADP]–ribose and ADP-ribose). Indeed, the enzymatic activities of CD157 in circulating neutrophils as well as in dimethyl sulfoxide (DMSO)–differentiated (CD157+/CD38-) HL-60 cells, are hardly detectable. This work also shows that the receptorial activity relies on cross-talk between CD157 and β2 integrin. CD157 localizes in GM1-enriched lipid rafts and, upon activation, it migrates to the uropod, a structure specialized in motility and adhesive functions. Indeed, CD157 is involved in adhesion to extracellular matrix proteins and in chemotaxis induced in vitro by formyl-methionyl-leucyl-phenylalanine (fMLP). These findings were consistent with the results obtained in neutrophils from patients with paroxysmal nocturnal hemoglobinuria (PNH), in which CD157 is deficient. These neutrophils showed constant defects in adhesion and migration. Our data attribute specific and crucial roles to CD157 in the regulation of innate immunity during inflammation.

scholarly journals Effects of enamel matrix proteins on adherence, proliferation and migration of epithelial cells: A real-time in vitro study

2016 ◽  
Vol 13 (1) ◽  
pp. 160-168 ◽  
Author(s):  
Marzena Wyganowska-Swiatkowska ◽  
Paulina Urbaniak ◽  
Daniel Lipinski ◽  
Marlena Szalata ◽  
Karolina Borysiak ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Real Time ◽  
In Vitro Study ◽  
Vitro Study ◽  
Matrix Proteins ◽  
Enamel Matrix ◽  
Enamel Matrix Proteins ◽  
And Migration ◽  
Proliferation And Migration

scholarly journals Antenatal betamethasone increases vascular reactivity to endothelin-1 by upregulation of CD38/cADPR signaling

2014 ◽  
Vol 5 (1) ◽  
pp. 56-62 ◽  
Author(s):  
J.-H. Lee ◽  
J. Zhang ◽  
G. A. Massmann ◽  
J. P. Figueroa
Keyword(s):  
Vascular Reactivity ◽  
Adult Life ◽  
Adenosine Diphosphate ◽  
Adp Ribosyl Cyclase ◽  
Antenatal Steroids ◽  
Pregnant Sheep ◽  
Antenatal Steroid ◽  
Increased Sensitivity ◽  
Cyclic Adenosine Diphosphate

Antenatal steroid administration is associated with hypertension in adult life; however, the mechanisms underlying this phenomenon are unclear. The aim of this study was to further characterize the effects of antenatal glucocorticoid exposure on the endothelin (ET-1) system, specifically to ascertain the role of the cyclic adenosine diphosphate ribose (cADPR)/ryanodine receptor pathway in the increased sensitivity to ET-1 observed in the offspring exposed to antenatal glucocorticoids. Pregnant sheep were randomly treated with betamethasone (Beta; 0.17 mg/kg) or vehicle at 80 and 81 days of gestation. In adults, we studied endothelium-denuded arterial segments of the brachial arteries. ET-1-induced vasoconstriction was significantly higher in the arteries from Beta sheep (F=3.5, P<0.05). Inhibition of ADP-ribosyl cyclase with 2-2'-dihydroxy-azobenzene significantly decreased the ET-1-induced contraction in Beta but not in vehicle-treated sheep. Nicotinamide attenuated ET-1 contraction in both, but it was significantly more pronounced in the Beta-treated sheep. No significant differences were observed following KCl-induced (6.25–75 mM) contraction. Nicotinamide (10 mM) significantly attenuated the KCl-induced vasoconstriction in both groups. In KCl (62.5 mM)-constricted arteries, the effect of nicotinamide (NIC) was significantly greater in the vehicle-treated sheep (50% relaxation v. 40% relaxation; t=2.2, P<0.05). In contrast, the sodium nitroprusside (SNP) relaxation was not statistically different. An additive effect was observed when NIC and SNP were used in combination and it was also more pronounced in vehicle-treated sheep. We conclude that the increased response to ET-1 is mediated by activation of the CD38/cADPR signaling pathway. Further studies are required to identify the effectors downstream from cADPR affected by exposure to antenatal steroids.

scholarly journals Urokinase Receptor (CD87) Regulates Leukocyte Recruitment via β2 Integrins In Vivo

1998 ◽  
Vol 188 (6) ◽  
pp. 1029-1037 ◽  
Author(s):  
Andreas E. May ◽  
Sandip M. Kanse ◽  
Leif R. Lund ◽  
Roland H. Gisler ◽  
Beat A. Imhof ◽  
...  
Keyword(s):  
Leukocyte Adhesion ◽  
Close Association ◽  
Phosphatidyl Inositol ◽  
Urokinase Receptor ◽  
Β2 Integrin ◽  
And Migration ◽  
Deficient Mice ◽  
Β2 Integrins

The urokinase receptor (CD87; uPAR) is found in close association with β2 integrins on leukocytes. We studied the functional consequence of this association for leukocyte adhesion and migration. In vivo, the β2 integrin–dependent recruitment of leukocytes to the inflamed peritoneum of uPAR-deficient mice was significantly reduced as compared with wild-type animals. In vitro, β2 integrin–mediated adhesion of leukocytes to endothelium was lost upon removal of uPAR from the leukocyte surface by phosphatidyl-inositol–specific phospholipase C. Leukocyte adhesion was reconstituted when soluble intact uPAR, but not a truncated form lacking the uPA-binding domain, was allowed to reassociate with the cell surface. uPAR ligation with a monoclonal antibody induced adhesion of monocytic cells and neutrophils to vascular endothelium by six- to eightfold, whereas ligation with inactivated uPA significantly reduced cell-to-cell adhesion irrespective of the β2 integrin–stimulating pathway. These data indicate that β2 integrin–mediated leukocyte–endothelial cell interactions and recruitment to inflamed areas require the presence of uPAR and define a new phenotype for uPAR-deficient mice. Moreover, uPAR ligation differentially modulates leukocyte adhesion to endothelium and provides novel targets for therapeutic strategies in inflammation-related vascular pathologies.

scholarly journals Insights into In Vitro Wound Closure on Two Biopolyesters—Polylactide and Polyhydroxyoctanoate

Materials ◽  
2020 ◽  
Vol 13 (12) ◽  
pp. 2793 ◽  
Author(s):  
Tomasz Witko ◽  
Daria Solarz ◽  
Karolina Feliksiak ◽  
Katarzyna Haraźna ◽  
Zenon Rajfur ◽  
...  
Keyword(s):  
Cell Shape ◽  
Wound Closure ◽  
Cellular Migration ◽  
Wound Dressings ◽  
Embryonic Fibroblasts ◽  
Cell Processes ◽  
Migration Strategies ◽  
And Migration ◽  
Microscopic Techniques

Two bio-based polymers have been compared in this study, namely: polylactide (PLA) and polyhydroxyoctanoate (PHO). Due to their properties such as biocompatibility, and biointegrity they are considered to be valuable materials for medical purposes, i.e., creating scaffolds or wound dressings. Presented biopolymers were investigated for their impact on cellular migration strategies of mouse embryonic fibroblasts (MEF) 3T3 cell line. Advanced microscopic techniques, including confocal microscopy and immunofluorescent protocols, enabled the thorough analysis of the cell shape and migration. Application of wound healing assay combined with dedicated software allowed us to perform quantitative analysis of wound closure dynamics. The outcome of the experiments demonstrated that the wound closure dynamics for PLA differs from PHO. Single fibroblasts grown on PLA moved 1.5-fold faster, than those migrating on the PHO surface. However, when a layer of cells was considered, the wound closure was by 4.1 h faster for PHO material. The accomplished work confirms the potential of PLA and PHO as excellent candidates for medical applications, due to their properties that propagate cell migration, vitality, and proliferation—essential cell processes in the healing of damaged tissues.

Platelet Adhesion to Collagen Is Inhibited by Adenosine Diphosphate but Unaffected by Cell Shape

1981 ◽  
Vol 46 (02) ◽  
pp. 515-520 ◽  
Author(s):  
F A Meyer ◽  
Z Weisman ◽  
M M Frojmovic
Keyword(s):  
Binding Sites ◽  
Cell Shape ◽  
Platelet Adhesion ◽  
Shape Change ◽  
Adenosine Diphosphate ◽  
Time Dependent ◽  
Rabbit Platelets ◽  
Normal Adhesion ◽  
Per Se

SummaryThe effect of limited platelet activation, in the absence of aggregation, on the subsequent ability of rabbit platelets to adhere to collagen was studied in vitro. ADP at concentrations that initiate shape change (>0.01. μM) reduce platelet adhesion. Shape change per se however, was not responsible since the time-dependence of the effect of ADP on shape change and adhesion is different and ADP induces reduced adhesion even when shape change is prevented with PGE1 or fixatives. Platelets shape changed without ADP addition, e. g. by chilling or by low ethanol concentrations, display normal adhesion to collagenIt appears likely that upon binding to the platelet ADP induces a time-dependent alteration in the membrane, akin to refractoriness, that influences binding sites for collagen. The effect of ADP can be blocked by prior addition of AMP but not if the additions are reversed. The implications of the present findings for platelet adhesion studies are discussed.

scholarly journals Regulation of LFA-1–dependent inflammatory cell recruitment by Cbl-b and 14-3-3 proteins

Blood ◽  
2008 ◽  
Vol 111 (7) ◽  
pp. 3607-3614 ◽  
Author(s):  
Eun Young Choi ◽  
Valeria V. Orlova ◽  
Susanna C. Fagerholm ◽  
Susanna M. Nurmi ◽  
Li Zhang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Inflammatory Cell ◽  
Mononuclear Phagocytes ◽  
Cell Recruitment ◽  
Β2 Integrin ◽  
Cytoplasmic Proteins ◽  
Inflammatory Cell Recruitment ◽  
And Migration

Abstract Inside-out signaling regulation of the β2-integrin leukocyte function–associated antigen-1 (LFA-1) by different cytoplasmic proteins, including 14-3-3 proteins, is essential for adhesion and migration of immune cells. Here, we identify a new pathway for the regulation of LFA-1 activity by Cbl-b, an adapter molecule and ubiquitin ligase that modulates several signaling pathways. Cbl-b−/− mice displayed increased macrophage recruitment in thioglycollate-induced peritonitis, which was attributed to Cbl-b deficiency in macrophages, as assessed by bone marrow chimera experiments. In vitro, Cbl-b−/− bone marrow–derived mononuclear phagocytes (BMDMs) displayed increased adhesion to endothelial cells. Activation of LFA-1 in Cbl-b–deficient cells was responsible for their increased endothelial adhesion in vitro and peritoneal recruitment in vivo, as the phenotype of Cbl-b deficiency was reversed in Cbl-b−/−LFA-1−/− mice. Consistently, LFA-1–mediated adhesion of BMDM to ICAM-1 but not VLA-4–mediated adhesion to VCAM-1 was enhanced by Cbl-b deficiency. Cbl-b deficiency resulted in increased phosphorylation of T758 in the β2-chain of LFA-1 and thereby in enhanced association of 14-3-3β protein with the β2-chain, leading to activation of LFA-1. Consistently, disruption of the 14-3-3/β2-integrin interaction abrogated the enhanced ICAM-1 adhesion of Cbl-b−/− BMDMs. In conclusion, Cbl-b deficiency activates LFA-1 and LFA-1–mediated inflammatory cell recruitment by stimulating the interaction between the LFA-1 β-chain and 14-3-3 proteins.

scholarly journals Stem Cell Factor Combined with Matrix Proteins Regulates the Attachment and Migration of Melanocyte Precursors of Human Hair Follicles in Vitro

2013 ◽  
Vol 36 (8) ◽  
pp. 1317-1325 ◽  
Author(s):  
Da-guang Wang ◽  
Xi-hui Xu ◽  
Hui-jun Ma ◽  
Cheng-rang Li ◽  
Xue-zhuang Yue ◽  
...  
Keyword(s):  
Stem Cell ◽  
Human Hair ◽  
Stem Cell Factor ◽  
Hair Follicles ◽  
Matrix Proteins ◽  
And Migration

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

1997 ◽  
Vol 10 (01) ◽  
pp. 6-11 ◽  
Author(s):  
R. F. Rosenbusch ◽  
L. C. Booth ◽  
L. A. Dahlgren
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Tendon Healing ◽  
Matrix Proteins ◽  
Isolated Cells ◽  
Superficial Digital Flexor Tendon ◽  
Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

Haemostatic Platelet Plug Formation in the Isolated Rabbit Mesenteric Preparation - An Analysis of Red Blood Cell Participation

1980 ◽  
Vol 44 (01) ◽  
pp. 006-008 ◽  
Author(s):  
D Bergqvist ◽  
K-E Arfors
Keyword(s):  
Whole Blood ◽  
Platelet Rich Plasma ◽  
Adenosine Diphosphate ◽  
In Vitro Test ◽  
Pharmacological Agents ◽  
Vitro Test ◽  
Releasing Effect ◽  
Plug Formation

SummaryIn a model using an isolated rabbit mesenteric preparation microvessels were transected and the time until haemostatic plugs formed was registered. Perfusion of platelet rich plasma gave no haemostasis whereas whole blood did. Addition of chlorpromazine or adenosine to the whole blood significantly prolonged the time for haemostasis, and addition of ADP to the platelet rich plasma significantly shortened it. It is concluded that red cells are necessary for a normal haemostasis in this model, probably by a combination of a haemodynamic and ADP releasing effect.The fundamental role of platelets in haemostatic plug formation is unquestionable but there are still problems concerning the stimulus for this process to start. Three platelet aggregating substances have been discussed – thrombin, adenosine diphosphate (ADP) and collagen. Evidence speaking in favour of thrombin is, however, very minimal, and the discussion has to be focused on collagen and ADP. In an in vitro system using polyethylene tubings we have shown that "haemostasis" can be obtained without the presence of collagen but against these results can be argued that it is only another in vitro test for platelet aggregation (1).To be able to induce haemostasis in this model, however, the presence of red blood cells is necessary. To further study this problem we have developed a model where haemostatic plug formation can be studied in the isolated rabbit mesentery and we have briefly reported on this (2).Thus, it is possible to perfuse the vessels with whole blood as well as with platelet rich plasma (PRP) and different pharmacological agents of importance.

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