Uncorrected Version. Published on October 23, 2008 as DOI:10.1189/jlb.


Molecular and functional interactions among monocytes, platelets, and endothelial cells and their relevance for cardiovascular diseases
Janine M. van Gils,* Jaap Jan Zwaginga,*,† and Peter L. Hordijk*,1 *Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands; and †Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands

Abstract: Platelets, monocytes, and endothelial cells are instrumental in the development and progression of cardiovascular diseases. Inflammation, a key process underlying cardiovascular disorders, is accompanied and amplified by activation of platelets and consequent binding of such platelets to the endothelium. There, platelet-derived chemokines, in conjunction with increased expression of adhesion molecules, promote the recruitment of circulating monocytes that will eventually migrate across the endothelial lining of the vessel into the tissues. Additionally, platelets may already become activated in the circulation and may form plateletmonocyte complexes, which show increased adhesive and migratory capacities themselves but also facilitate recruitment of noncomplexed leukocytes. They should therefore be considered as important mediators of inflammation. In molecular terms, these events are additionally governed by chemokines released and presented by the endothelium as well as the different classes of endothelial adhesion molecules that regulate the interactions among the various cell types. Most important in this respect are the selectins and their ligands, such as P-selectin glycoprotein (GP) ligand 1, and the integrins binding to Ig-like cell adhesion molecules as well as to GP, such as von Willebrand factor, present in the extracellular matrix or on activated endothelium. This review aims to provide an overview of these complex interactions and of their functional implications for inflammation and development of cardiovascular disease. J. Leukoc. Biol. 85: 000 – 000; 2009.
Key Words: inflammation ⅐ adhesion ⅐ endothelium

cells (EC) that line the intima, the innermost layer of the vessel wall. In addition, the inflammatory response is characterized by the accumulation of inflammatory cells in the intima, thus initiating the atherogenic process [1, 2]. The role of platelets in atherosclerosis was initially only believed to be in thrombus formation upon rupture of the more developed atherosclerotic plaques. Now, platelets are also known to assist and modulate inflammatory reactions and immune responses [3]. Therefore, platelets, together with inflammatory cells, are regarded as important players by integrating inflammatory responses, thrombosis, and atherogenesis. Activated platelets, adhered to a damaged vessel wall or to activated endothelium, have been shown to further promote local recruitment of leukocytes. As monocyte adhesion to the vascular wall, transendothelial migration, and differentiation toward macrophages are critical for the formation of atherosclerotic lesions, it is important to realize that these events are subject to regulation by platelet adhesion molecules and platelet-derived chemokines and cytokines. Platelet activation results in an increase in circulating leukocyte-platelet aggregates. In particular, platelet-monocyte complexes (PMC) have been observed in clinical conditions such as peripheral vascular disease, hypertension [4], acute or stable coronary syndromes [5–7], stroke [8], or diabetes [9]. Increased levels of PMC are also an early marker of acute myocardial infarction [7]. Conversely, high dietary intake of omega-3 fatty acids induces a reduction in activated platelets and PMC level [10]. However, the presence of PMC is not just a sensitive marker for in vivo platelet activation and cardiovascular diseases but is also regarded more and more as a cardiovascular risk factor [8, 11]. The importance of activated platelets and PMC in vascular disease is underscored by several studies that showed that prevention of platelet adhesion to monocytes by interfering with the binding of platelet P-selectin to P-selectin glycoprotein (GP) ligand 1 (PSGL-1) reduces inflammation. Infusion of

Cardiovascular diseases are the number one cause of death in the Western world and are predicted to remain so. Atherosclerosis, the primary cause of cardiovascular disease, is a systemic inflammatory disease. The inflammatory nature of atherosclerosis involves chronic stimulation of the endothelial
0741-5400/09/0085-0001 © Society for Leukocyte Biology
Correspondence: Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands. E-mail: p.hordijk@sanquin.nl Received July 6, 2008; revised September 1, 2008; accepted September 2, 2008. doi: 10.1189/jlb.0708400

Journal of Leukocyte Biology Volume 85, January 2009


Copyright 2008 by The Society for Leukocyte Biology.

1. 30] (Fig. RANTES contributes to smooth muscle cell proliferation [19]. the integrins ␣IIb␤3 (GPIIbIIIa. such as collagen and VWF. PMC are not merely a reflection of platelet activation. The initial contact by GPIb-IX-V and GPVI binding to VWF and collagen. The requirement for ␣IIb␤3 in mediating firm adhesion of platelets to the endothelium was shown by using plate- PLATELET ACTIVATION AND ADHESION TO THE VASCULAR WALL Molecular ligands The function of blood platelets is to arrest bleeding (hemostasis) by formation of stable blood clots following activation of the coagulation cascade. 15]. respectively. 31]. http://www. TNF superfamiliy 14 (TNFSF14)–TNFSF14R. Platelet– endothelial molecular interactions. by amplifying monocyte adhesion and migration and by promoting monocyte differentiation toward macrophages. but in view of the above. However. As a result. nonactivated endothelium normally prevents adhesion of platelets to the vessel wall by its antithrombotic properties. 32] (Fig. 1).org . ␣v␤3 integrin. or endothelial adhesion molecules or indirectly via additional bridging molecules. Upon activation of integrins. the platelet chemokines platelet factor 4 (PF4) and RANTES contribute to lesion progression by inducing monocyte survival and differentiation into macrophages [17]. Increased levels of PMC in patients with cardiovascular disease have so far been regarded as a parameterreflecting disease. but platelet binding also leads to an activated and thus more proatherogenic monocyte phenotype. ECM. These activated integrins are required and essential for stable platelet adhesion to the ECM and EC. fibrinogen receptor) and ␣2␤1 (collagen receptor) are activated [26. respectively [30. and EC and the consequences of these interactions for the development of cardiovascular diseases as well as possibilities for intervention. 13]. and GPIb are involved in the tethering and rolling of platelets on the EC. P-selectin and GPIb can mediate rolling interactions between platelets that are still in the circulation and those that are already adhered to the vessel wall. PMC also seem able to play a key role in disease pathogenesis. Finally. Additionally. platelets may contribute to the integrity of the endothelium [20] and participate in inflammatory processes [21]. even under high shearstress conditions [24. platelet adhesion at sites of vascular injury involves initial tethering and rolling 2 Journal of Leukocyte Biology Volume 85. fibronectin. Also. All of these data indicate a role of P-selectin–PSGL-1 interactions in atherosclerosis. monocytes.jleukbio. 1).human recombinant (r)PSGL-1 in animal models of vascular injury preserved vascular endothelial function [12. Furthermore. VWF especially mediates direct interaction of platelets with intact. and cyclooxygenase-2 [22]. involving release of platelet activation-inhibiting substances such as NO. also known as the VWF receptor complex. infusion of activated but not of Pselectin-deficient platelets results in increased formation of atherosclerotic lesions [16]. The VWF–GPIb-IX interaction has been shown to induce Syk phosphorylation and ␣IIb␤3 integrin activation [31]. The ligand-receptor combinations P-selectin. This process is mediated by adhesion to VWF via the membrane adhesion receptor GP complex Ib-IX-V. and to collagen via GPVI [26. results in platelet activation via a complex series of intracellular reactions. VWF. prostacyclin. In addition. Rolling on intact endothelium is also mediated by binding of GPIb to P-selectin on EC [25. CD40L and TNFSF14 binding to the endothelial ligands induces an inflammatory response in the EC. The latter involves plateletbound fibrinogen. January 2009 Fig. This can be through direct binding of the integrins to collagen. platelets activated already can tether and roll on PSGL-1 and GPIb on activated EC even under high shear [29. PSGL-1. fibrinogen (Fg). Stable adhesion. mediating progression to a fibrous plaque [2]. activated EC. also CD40 ligand (CD40L)–CD40. Upon firm adhesion. not only by inducing expression and secretion of cytokines and active substances from platelets and monocytes but most importantly. This review will discuss in more detail the molecular mechanisms involved in cell– cell interactions among platelets. Schematic overview of the various interactions between endothelial and platelet surface molecules. Platelet adherence is even more stimulated upon vessel wall damage when extracellular matrix (ECM) proteins are exposed. 28]. and GPIb. the endothelial phenotype can change to prothrombotic by release of platelet-binding and stimulating agents such as ADP and multimeric Von Willebrand factor (VWF) and the up-regulation of expression of tissue factor (TF) and of adhesion molecules [23]. and junctional adhesion molecule A (JAM-A)–JAM-A interactions are initiated. promotes foam cell development [18]. Rapid platelet adhesion to the ECM followed by their activation is the primary event in thrombus formation. requires additional contacts between the platelets and the ECM or the endothelium (Fig. which bind to endothelial ICAM-1. Clearly. 27]. are strong ligands of platelet GP. 25]. over the ECM and intact endothelium. PF4 also facilitates the esterification and promotes the uptake of oxidized lowdensity lipoprotein by macrophages and thereby. Healthy. Moreover. 1). Under physiological flow conditions. platelets firmly adhere to the endothelium. and VWF. and VWF. in an inflamed vessel wall. the absence of P-selectin in mice diminishes lesion formation [14. mainly via the additional bridging molecules fibronectin (Fn). however. Additionally.

In conclusion. and NF-␬B activation [39. PF4 drastically enhances RANTES-induced monocyte arrest on EC [58]. and endothelial cells 3 . JAM-A and platelet-associated TNFSF14 (also known as LIGHT. VWF. constitutively expressed on monocytes [46]. Besides PSGL-1. reflected by spreading and increased surface expression of adhesion molecules. 40]. and MMPs is induced by platelet binding (4). but also by secretion of potent inflammatory substances. E-selectin expression via activation of the NF-␬B pathway is also induced by platelet-released PF4 [44]. which further stimulate their biological activity. 34]. Furthermore. fibrinogen is linked to Mac-1 and its platelet surface counterpart GPIIbIIIa [32]. For instance. ligation of platelet P-selectin rapidly stimulates Weibel-Palade body release. 42].lets defective in ␣IIb␤3 or by adding ␤3-integrin antagonists or a blocking antibody [33. The endothelial deposition of plateletderived RANTES has been shown to trigger further monocyte arrest on the endothelium under high shear but not on endothelium-adherent platelets [19]. 55–57]. such as CD40L. 3). expression of adhesion molecules. similar bridging mechanisms as described above for platelets and EC also mediate plateletmonocyte binding. such as IL-1␤ and PF4 [37. also. Also. Mac-1 on leukocytes binds to GPIb [49] and to JAM-C on platelets [50]. 2). Subsequently. in mice lacking ICAM-1. which itself supports interactions with platelets. Activation of EC by adhesion of platelets Stable binding of platelets to the endothelium or to ECM results in strong activation of these platelets. platelet adhesion to activated EC is strongly reduced. P-selectin expression on the endothelium [45]. and P-selectin. predominantly mediated by CCR1 [59]. respectively [16. Monocytes. and production of reactive oxygen species (ROS) [41– 43]. 2. In conclusion. For example. Next to adhesion molecules. Intracellular signaling induced upon platelet binding results in NF-␬B activity and ROS production (2). the chemokines plateletactivating factor and MIP are secreted by platelets adhered on the endothelium. platelet adhesion endows the endothelium with a proinflammatory phenotype (Fig. Conversely. 38]. Rolling is mediated by P-selectin on activated platelets and PMC—FORMATION AND FUNCTIONAL CONSEQUENCES Although a rare event under resting conditions. and IL-6 secretion. van Gils et al. resulting in. and matrix metalloproteinase (MMP)-2 and -9. platelet adhesion to the EC or ECM and chemokines secreted by platelets greatly contribute to subsequent monocyte adhesion to the vascular wall. On monocytes. Additional interactions between platelets and monocytes include CD40L–CD40 [52] and monocyte triggering receptor expressed on myeloid cell 1 (TREM-1) to platelet-expressed TREM-1 ligand [53. including increased secretion of IL-8 and MCP-1. 1). The deposited platelet chemokines form homophilic as well as heterophilic aggregates. Besides direct interaction. Thus. reflected by up-regulation of adhesion molecules (E-selectin and VCAM-1) and release of chemokines (MCP-1 and IL-8) [38]. GMCSF. Also. bridging by thrombospondin of the CD36 antigens (present on monocytes and platelets) was shown [51]. a series of adhesion molecules is up-regulated (3). CD40L (CD154) is stored in high amounts and released by platelets within seconds after GPIIbIIIa ligation [41. known to signal inflammatory reactions within EC. RANTES increases the binding of PF4 to the monocyte surface [58]. Furthermore. 54] (Fig. platelets. urokinasetype plasminogen activator (uPAR). Finally. The initial association between platelet P-selectin and monocyte PSGL-1 leads to increased expression of the ␤2-integrin CD11b/CD18 [␣M␤2. Also. membrane-activated complex 1 (Mac-1)] on the monocytes [48]. Different mechanisms Fig. CD15 (Lewis X) on monocytes has also been shown to bind platelet P-selectin [47]. multiple interactions between surface receptors on EC and platelets result in firm adhesion of platelets at sites of vascular injury. TNFSF14. 36] (Fig. chemokines deposited on the endothelium are facilitating recruitment of monocytes. Adhered platelets efficiently mediate monocyte rolling and arrest. next to VWF release. and secretion of several cytokines. even at high shear. ICAM-1 and ␣v␤3 integrin expression. Platelets adherent to EC recruit monocytes Atherosclerosis is characterized by monocyte and macrophage accumulation in the vascular intima. identified in ADP-stimulated platelets) contribute to firm adhesion of platelets to the endothelium [35. RANTES and PF4 can be deposited on EC by activated platelets or platelet microparticles upon adhesion or even during transient interaction through JAM-A or P-selectin. platelets in the circulation sometimes do get activated. PSGL-1. inducing MCP-1. TNFSF14 can induce an inflammatory response in EC. IL-1␤ is synthesized and released by platelets in significant amounts and has been identified as a key mediator of platelet-induced activation of EC. such as RANTES and PF4 (1). Endothelial activation upon platelet binding. Platelet interaction with EC mediates deposition of platelet-derived chemokines. This results in stimulation of EC through the cognate receptor CD40.

could be responsible for the activation of circulating platelets. Michelson et al. http://www. Steroid receptor coactivator. The link between human monocyte subsets and formation of PMC therefore still needs further clarification. protein kinase C. 67]. Monocyte expression of chemokines. for instance. TF expression by the monocytes is reduced by a P-selectinblocking antibody and by IL-10. to its receptor on monocytes. Furthermore. augments ␤2-integrin avidity upon PSGL-1 cross-linking [78]. Platelets bind via P-selectin. indeed had a short circulation time. in patients with percutaneous coronary intervention. The ligation of PSGL-1 induces integrin activation on monocytes. by released agents from platelets from unstable thrombi [60]. which to monocytes leads to the deposition of platelet-derived chemokines. mediated by additional bridging molecules. This could contribute to the role of monocytes in atherogenesis. no difference in PSGL-1 expression has been detected [66.jleukbio. Antibody inhibition studies indicate that the platelet-monocyte conjugation is abolished by blocking P-selectin and partially inhibited by other blocking antibodies [49. PSGL-1 [61]. The in vivo circulation time and clearance of the complexes formed between activated platelets and monocytes are also not yet well-defined. January 2009 The binding of platelets to monocytes mediated via P-selectin– PSGL-1 interactions induces L-selectin shedding from the monocyte surface [75] (Fig. Monocyte activation upon platelet binding. In vivo.org . Monocyte activation upon platelet interaction Fig. Src. conditions such as systemic inflammation and acute myocardial infarction increase the number of activated platelets in the circulation. shown to be increased dramatically in hypercholesterolemic mice [63. 71. by cytokines associated with systemic thromboembolic or inflammatory events. engagement of CD40 with CD40L. PMC were detected much longer and only returned to baseline after 24 h [74]. demonstrates a higher expression of PSGL-1 [65].g. 64]. in a study using primates. 79 – 81]. but not by a CD40L antibody [52. This 4 Journal of Leukocyte Biology Volume 85. Huo et al. induced by Fig. or by rolling interactions with activated endothelium. and CD36 –CD36 via thrombospondin. 4. in part. but also TREM-1 ligation. TREM-1 ligand–TREM-1. 69]. and contradicting data have also been presented [68.indicates that platelet P-selectin is the critical ligand initiating PMC formation via binding to PSGL-1. deposited by platelets onto the monocytes. results in an increase in monocyte adhesive capacity by up-regulation of ␤1. The increased adhesive capacity of these complexes is likely to have a major influence on their clearance. but monocytes seem most proficient in this and are therefore the focus of our review. 4).. and other ligands play only an additive role. P-selectin mediates the initial binding contact with monocytes via PSGL-1 and CD15. e. These authors also showed that the PMC were cleared by monocyte transmigration. 76]. Reports about this matter are only few. results in L-selectin shedding (2) and increased expression and activation of ␤1. 61. PKC. [74] found that the lifespan of PMC was not related to platelet P-selectin shedding. which increases the level of activated platelets. Furthermore. 3. These activated platelets are able to bind to all types of leukocytes. Intracellular signaling induced upon platelet binding via PSGL-1.and ␤2-integrins [54. although so far in human monocyte subsets. However. resulting in further binding interactions. The platelet-binding capacity between the different subsets of monocytes is still unknown. The extent of PMC formation is mostly dependent on platelet activation [61] and to a limited extentl. which may also promote integrin expression. Whatever the cause. Monocyte binding to activated platelets has also been shown to increase the production of various proinflammatory mediators and TF expression. by turbulent flow. The presence of the chemokines RANTES and CXCL10. In mice. However. expressed on the surface of activated platelets. [16] have shown in mice that PMC. 72]. Further interactions are through CD40L–CD40.and ␤2-integrins (3). P-selectin is expressed upon platelet activation for several hours before it is shed from the surface [73]. fagocytic uptake of the platelets by monocytes might also contribute to a reduction in PMC levels. on monocyte activation [70]. this interaction between platelets and monocytes was found to increase expression and activity of the ␣4␤1 and ␣M␤2-integrins [75. Similarly. Comparing monocytes with neutrophils in this respect showed more and initially faster binding of activated platelets to monocytes [61. Platelet–monocyte molecular interactions. 77]. such as RANTES and PF4 (1). Finally. formed upon injection of activated platelets. the inflammatory monocyte Ly-6Chi subset. P-selectin–PSGL-1 interactions are important but not exclusively responsible for these processes. 62]. monocyte NF-␬B activity (4) and the secretion of several cytokines and TF (5) are induced upon binding of activated platelets.

The activation of mTOR is essential for the transcription and synthesis of uPAR and Rho kinase 1 [94. result in more monocyte adhesion upon platelet binding to the monocytes. lymphocytes. and IL-8 [54. Rolling is mediated by monocyte-expressed L-selectin and endothelialexpressed P. Syk. 95]. 5. shown in lymphocytes [78] (Fig. however. adhesion.or L-selectin [98]. TNF-␣. PSGL-1 ligation induces production of superoxide anion radicals from monocytes and neutrophils [85]. ERK. 94]. 100]. adhesion. 5). van Gils et al. initiating the atherogenic program.thrombin-activated platelets. Secondary tethering represents monocyte adhesion to other already-adhered monocytes [99. Also PKC isoforms are activated. Src kinase. activation of GTPase Ras as shown in neutrophils [86]. Chronic activation of EC results in increased Fig. together with the L-selectin shedding. activation. which is important for the activation of LFA-1 (␣L␤2) integrins [92] and for the induction of serum response element-dependent transcriptional activity [87]. Ligation of TREM-1 or the ligation of monocyte PSGL-1 together with RANTES. platelets. 102]. although PF4 has been shown to induce the secretion of TNF-␣ by monocytes as well [17]. was found to bind to the cytoplasmic domain of PSGL-1. a novel protein selectin ligand interactor cytoplasmic 1 (SLIC-1). is regulated by NF-␬B activity [82]. The ERM proteins also mediate PSGL-1 association with Syk [87]. platelet binding alters the adhesive and migratory phenotype of monocytes. Monocytes. demonstrated in neutrophils. SLIC-1 serves as a sorting molecule that promotes traffic of PSGL-1 to endosomes [96]. Furthermore. Platelet binding to monocytes also results in shedding of L-selectin from the monocyte surface [75]. PMC show increased primary and secondary tethering on EC and on already-adhered inflammatory cells [101. resulting in firm. chemoattractant signals and increased expression and activity of various adhesion molecules on the cell surface that mediate adhesion to and migration across the endothelium of inflammatory cells. The inflammatory nature of atherosclerosis involves chronic stimulation of the EC by lipids in the intima [1–3]. and paxillin. induce activation of monocytes. stable adhesion to the EC through binding to ICAM-1. Rolling triggers further expression and activation of monocyte integrins. which are both involved in adhesion and migration processes. 84]. CD44. Taken together. plateletderived chemokines. or E-selectin-ligand-1 (ESL-1) [97] (Fig. As described above. mediating integrin activation. interacting with PSGL-1. is not only an adhesion but also a signaling molecule. which is crucial for leukocyte rolling [91]. and various monocytic cellular models [86 – 89]. 83. Primary tethering occurs directly at the endothelial surface. 4).and E-selectin. and transmigration. The cytoplasmic tail of PSGL-1 is linked to the actin cytoskeleton through the ezrin-radixin-moesin (ERM) proteins [90]. decreasing the rolling velocity of activated monocytes. Tethering and rolling Similar to the interactions between platelets and EC under physiological flow. resulting in changes in expression of adhesion molecules and secretion of cytokines (Fig. or ␣v␤3 via fibronectin. PSGL-1. CD44 is subsequently important for reducing the rolling velocity of leukocytes after they have tethered through P. such as pp125 focal adhesion kinase. which has no apparent signaling role upon leukocyte adhesion. 4). PMC ADHESION AND TRANSENDOTHELIAL MIGRATION The migration of monocytes across the vascular endothelium is required for immune surveillance and for monocyte recruitment at inflammatory sites. rolling. and tyrosine phosphorylation of various cytoplasmic proteins. Monocyte extravasation is tightly regulated by a multi-step process of tethering. Recently. the cytoplasmic tail of PSGL-1 also interacts with Nef-associated factor 1 (Naf1) [89]. induced by PSGL-1 in rolling. VCAM-1. Upon PSGL-1 engagement. Monocyte–EC molecular interactions. and migration of monocytes during inflammatory responses. monocyte adhesion to the vessel wall also involves tethering and rolling over the endothelium. leading to activation of ␤2integrins. which results in activation of Akt and mammalian target of rapamycin (mTOR) [89. Naf1 is phosphorylated via Src kinase. The initial interaction of monocytes with the endothelium is by tethering and rolling. PSGL-1 and ESL-1 are primarily responsible for tethering and rolling of leukocytes on the endothelium. induces NF-␬B activity and subsequently secretion of MCP-1. These findings emphasize a critical role for intracellular signaling. PSGL-1 signaling PSGL-1 plays a major role in the binding of monocytes to activated platelets. and endothelial cells 5 . together with the ligation of various adhesion molecules on the monocyte following the interaction with activated platelets. Two types of monocyte tethering can be distinguished. but not PF4. The increased tethering and rolling. The Naf1-binding sites in the PSGL-1 cytoplasmic domain are distinct from the residues critical for the recognition of ERM proteins [93]. This is mediated by the selectins and their ligands.

124. However. favoring the probability of monocytes to encounter and to be activated by chemokines or lipid mediators presented on the endothelial surface [103]. 113]. chemokines from the underlying intima stimulate them to migrate through the endothelial monolayer into the subendothelial space. Blockade of JAM-B/-C decreased the 6 Journal of Leukocyte Biology Volume 85. 130. Abciximab did not significantly reduce the formation of PMC [126]. Clopidogrel greatly reduces PMC in patients with atherosclerotic diseases and has been shown to reduce P-selectin expression and CD40L release [122–124]. Rac1. in contrast to targeting platelet aggregation. 131]. is a good candidate for a future drug. In addition. PMC represent a potential therapeutic target for limiting cardiovascular diseases. adherent monocytes interact. Leukocyte arrest is induced further by leukocyte integrins ␣L␤2 (LFA-1) or ␣M␤2 (Mac-1) and VLA-4 ligation by the endothelial Ig superfamily members ICAM-1 and VCAM-1. revealing a role for these adhesion molecules in leukocyte recruitment. Leukocyte adhesion and ligation of ICAM-1 and VCAM-1 and the subsequent increase in endothelial actin stress-fiber formation and monolayer permeability are controlled by the GTPases RhoA. During this process. that show no or very little effect on PMC formation [122. Bradfield et al. induce a rapid increase in the binding affinity and avidity of ␤2-integrins of the leukocytes [104.and ␤1-integrins. The highaffinity binding of chemokines to specific G-protein-coupled receptors initiates the intracellular signaling cascade from these receptors to phospholipase C signaling. 128]. CD99. number of monocytes in the extravascular compartment by allowing multiple reverse-transmigration events. The EC participate actively in the transmigration event. much evidence points to an efficient inhibition of PMC formation by clopidogrel. Targeting inhibition of proinflammatory platelet activation or interaction. and JAMs also actively mediate leukocyte transendothelial migration through homophilic interactions [116 –118]. 121]. Furthermore. as well as angiogenesis [110. Use of rPSGL-1 in animal models indeed results in reduced platelet and leukocyte adhesion to the endothelium and better vascular function after injury [12. In contrast. Pselectin and PSGL-1 are logical. such as ICAM-1 and VCAM-1. platelets. the increased http://www. activation of small GTPases (Rap1). it can be concluded that platelet binding to monocytes results in increased monocyte adhesion and transmigration and subsequent platelet deposition on the endothelium. respectively [107] (Fig. 5).jleukbio. Importantly. JAM-A and JAM-C in various inflammation-related models. including clopidogrel (inhibition of ADP-mediated platelet activation) and Abciximab (GPIIbIIIa antibody). Although some studies suggest otherwise by reporting an increase in the expression of RANTES upon clopidogrel administration [125]. RANTES receptor antagonists inhibit the infiltration of monocytes and limit atherosclerotic plaque formation in proatherogenic mice models [55. [119] discovered a novel role for endothelial JAM-C in mice in regulating monocyte retention in the abluminal compartment after primary transmigration in vivo. and MCP-1 secreted by platelets and EC trigger arrest of rolling monocytes on EC [19. neointimal lesion formation. supporting adhesion of PMC. and receptors involved. are concentrated [114]. with JAM family members at the most apical regions of the interendothelial junctions. Thus. One of the results is a pronounced morphological response of the EC by forming “docking structures” [114] or “transmigratory cups” [115]. are shed from the monocyte as a result of monocytic PSGL-1 redistribution and mechanical stress. and transitional changes in integrin conformation through the association with actin-binding proteins. potential targets for intervention with antibodies or recombinant proteins. RANTES. which leads to stable adhesion [110]. 13. the cell– cell junctions disengage transiently and locally to allow the leukocyte to cross [112. 129]. another platelet aggregation inhibitor. in particular. Moreover. In these structures. Although Abciximab resulted in vitro in less platelet binding to monocytes and a decrease in TF expression on monocytes. On monocytes. and Rap1 in the EC [112].org . IL-8. 120]. PMC have induced integrin expression and activity compared with platelet-free monocytes. integrin ligands. The involvement of the JAMs in endothelial permeability and monocyte adhesion and transmigration suggests a broad relevance for JAMs in vascular inflammation. chemokines. Rolling and adhesion of leukocytes over activated endothelium are accompanied by a complex response from the EC. via their ␤2. involving extensive reorganization of the endothelial actin cytoskeleton and the activation of intracellular signaling pathways. As traditional platelet activation inhibitors show varying success in preventing PMC formation. monocytes. as shown in lymphocytes and monocytes [107–109]. there are some studies with aspirin. PMC show increased transmigration compared with plateletfree monocytes [75. This is corroborated further by a large number of studies that have established a role for. January 2009 Conclusions and future considerations Atherosclerosis and cardiovascular disease involve multifactorial mechanisms with interactions among coagulation. Also. increasing monocyte adhesion and transmigration capacity. targeting CD40L or RANTES may be beneficial. 106]. JAM-A also contributes to monocyte adhesion to atherosclerotic endothelium through its binding to LFA-1 [111] and indirectly through its homophilic binding to platelet JAM-A. Transmigration Upon binding of monocytes to the vessel wall. We have observed that the platelets do not remain attached to the monocyte following transmigration [121] but instead. and EC with multiple adhesion molecules. 105]. 127]. the ␣4␤1 (VLA-4) integrin is known to further slow the selectin ligand-dependent rolling. in cooperation with PSGL-1 ligation to endothelial and platelet ligands. chemokines on the luminal-endothelial surface. During transendothelial migration.Monocyte activation and firm adhesion Low velocities of rolling increase monocyte transit time through inflamed vessels. no effects or even an increase in PMC levels are observed [126. The junctional adhesion receptors PECAM-1. INTERVENTION POSSIBILITIES A number of therapeutic molecules have been used to investigate the inhibition of PMC.

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