LEADING ARTICLE

Activation of Adhesion Molecules in Septic Shock Patients

Marc Leone and Claude Martin
Intensive Care Unit and Trauma Center, Marseilles University Hospital System, Marseilles School of Medicine, Marseilles, France

The endothelium has been identified as the central effector in the inflammatory response. Adhesion molecules are strongly involved in the inflammatory process by modulating leukocyte trafficking. The most important adhesion molecules are the selectins (E-, L-, and P-selectins) and members of the immunoglobulin superfamily (intercellular adhesion molecule-1 and vascular cell adhesion molecule-1). Plasma levels of these molecules are increased in septic shock patients, which may be related to a marked activation of the endothelium. However, a dichotomous profile is observed between plasma and tissue expression. Current studies suggest that the expression of these molecules provides evidence for endothelial activation in septic shock patients, and that their modulation should make it possible to control the inflammatory response. The development of selective immunotherapy to inhibit the actions of adhesion molecules may hold the promise of limiting tissue injury in septic shock.
Systemic inflammatory response syndrome (SIRS) is a response to a variety of tissue insults, including sepsis and trauma. The pathogenesis of these critical illnesses is associated with marked changes in hemodynamics, organ blood flow, and immune function [1]. Sepsis results in endothelial activation and damage, and eventually multiple organ dysfunction (MOD) and death. The role of the endothelium through endothelial activation is probably crucial. During severe sepsis and septic shock [2], the lipopolysaccharide (LPS) released from the bacterial wall is believed to be central to the manifestation of shock. LPS is a potent inducer of cytokine release [3,4]. Frequent observations, including the association between the presence of tumor necrosis factor (TNF) in plasma and fatality in septic shock patients, have confirmed that cytokines are markers of severity and major mediators involved in the pathogenesis of septic shock. One of the most important targets of LPS and cytokine actions are the blood vessels, which undergo several structural and
Address for correspondence: Marc Leone, Hpital Nord, Dpartement dAnesthsie-Ranimation, 13915 Marseille Cedex 20, France. E-mail: Marc.Leone@ap-hm.fr

functional modifications that result in endothelial activation [35]. The activated endothelium expresses adhesion receptors, which control leukocyte trafficking at the site of inflammation. Endothelialleukocyte adhesion is one of the most important events in the inflammatory process. In the first phase, selectins mediate initial tethering to the vessel wall. This initial tethering makes it possible for leukocytes to roll along the blood vessel wall and to sense activating factors, such as chemokines, that are present on the surface of endothelial cells (ECs). This leads to activation of leukocyte integrins that belong to the immunoglobulin (Ig) superfamily and mediate firm adhesion, which is required to achieve the migration of leukocytes through ECs. Finally, they allow the leukocytes to transmigrate to the site of inflammation. The identification of specific molecules that mediated cell adhesion ushered in a new era in our understanding of the inflammatory process. The purpose of this review is to dissect the role of adhesion molecules in the context of inflammatory disease, discuss the profile that is observed in septic shock patients, and outline areas of future research that may have therapeutic implications.

56

ADVANCES IN SEPSIS Vol 2 No 2 2002

ACTIVATION OF ADHESION MOLECULES IN SEPTIC SHOCK PATIENTS

Pathophysiology Role of selectins The selectin family is comprised of three proteins designated by the prefixes L (leukocyte), P (platelet), and E (endothelial). It represents a class of cell adhesion molecules that are specialized for recruiting leukocytes from the flowing bloodstream. Their distribution is restricted to the leukocyte-vascular system. P-selectin, which is stored in the Weibel-Palade bodies of ECs, is rapidly mobilized to the plasma membrane in response to proinflammatory mediators. Cloning of P-selectin revealed the selectin nature of this molecule [6], and stimulated experiments that demonstrated the ability of P-selectin to mediate neutrophil binding to platelets and to ECs [7]. P-selectindeficient mice exhibit a number of defects in leukocyte behavior, including elevated numbers of circulating neutrophils, virtually total absence of leukocyte rolling in mesenteric venules, and delayed recruitment of neutrophils to the peritoneal cavity upon experimentally induced inflammation. Hence, P-selectin has a role in the early steps of leukocyte recruitment at sites of inflammation. In vivo experiments provided evidence that P-selectin glycoprotein ligand-1 is required for these early steps in the inflammatory response, although it is not required for cytokine-induced, E-selectin-mediated leukocyte rolling [8]. In addition, soluble P-selectins (sP-selectins) constitute an endogenous activator of the coagulation process through the generation of circulating microparticles in plasma. High levels of sP-selectins observed in thrombotic consumptive platelet disorders such as disseminated intravascular coagulation could be, in part, responsible for the generalized hypercoagulable state associate with this disease [9]. In addition, sP-selectins could contribute to the formation of fibrin tails [9]. In one variant, E-selectin is induced on vascular ECs by cytokines, and requires de novo mRNA and protein synthesis. E-selectin is mediated by TNF expression in most vascular beds during endotoxemia [10]. In E-selectindeficient mice, the percentage of circulating neutrophils was significantly higher than in other genotypes. E-selectin mediates granulocyte rolling at a significantly lower velocity than either L-selectin or P-selectin. Slow leukocyte rolling, as mediated by E-selectin, greatly increases the transit time of leukocytes through the microvascular network. This slowing down may serve to locally target an inflammatory response and may promote the transition from rolling to firm adhesion more rapidly after leukocytes encounter a chemoattractant. L-selectins are an important determinant for the penetration of leukocytes into tissue. They are involved

in the migration of neutrophils into inflamed tissue [11], and are expressed on most types of leukocytes. In fact, L-selectins are one of the first mediators of leukocyte rolling. A recombinant fusion protein carrying the extracellular portion of mouse L-selectin and the Fc portion of human IgG1 blocked leukocyte rolling in rat mesenteric venules [12]. An animal study provided evidence that L-selectin has a critical role in enhancing the ability of leukocytes to respond effectively to chemotactic stimuli. For instance, the response of leukocytes to platelet activating factor (PAF) was impaired in L-selectin-deficient-mice. This dysfunction can be explained by a reduced chemokinesis. These findings provide evidence that L-selectins facilitate leukocyte emigration and extravascular locomotion [13]. In mice deficient in E- and P-selectin, severe defects are observed in early leukocyte rolling and emigration of neutrophils. Hence, in models of acute inflammation, a considerable overlap in the functions of E- and P-selectin is highlighted [14]. These mice have an increased susceptibility to spontaneous infection over time [15]. Role of integrins The second step in leukocyte migration is through firm adhesion of leukocyte integrins with endothelial Ig superfamily members, of which there are >70. The roles of these family members are multiple, but are linked by the common theme of controlling cell behavior. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are among the most important ligands for leukocyte integrins. They are widely distributed on ECs and their surface expression is increased by cytokine-induced de novo protein synthesis. After activation, some adhesion molecules are also released into plasma as soluble forms with no cytoplasmic anchor sequence, either because of shedding from the ECs surface or differential mRNA splicing [16]. ICAM-1 is expressed in significant amounts on a limited number of cell types, including ECs, and is widely inducible, or up-regulated by interferon-g (IFN-g), interleukin-1b (IL-1b), TNF, and LPS. ICAM-2 is present on resting ECs. The high levels of ICAM-2 suggest that it is the major ligand for lymphocyte function-associated antigen-1 and, as such, may have a role in normal lymphocyte circulation. Mice deficient in ICAM-1 exhibit prominent abnormalities of inflammatory response including impaired neutrophil migration [17]. To define the cascade of interactions among families of adhesion molecules during leukocyteEC interactions in vivo, mice deficient in both L-selectin and ICAM-1 were generated

ADVANCES IN SEPSIS Vol 2 No 2 2002

57

MARC LEONE AND CLAUDE MARTIN

Table 1. Phenotypes of adhesion molecule-deficient mice.

Deficiency P-selectin E-selectin L-selectin P- and E-selectins ICAM-1 VCAM-1 P-selectin/ICAM-1 Health status Normal Normal Normal Spontaneous infection Normal Lethal Normal Neutrophil count Increased Normal Normal Increased Increased NA Increased Leukocyte rolling Absent Defective Reduced Absent NK NA NK Early extravasation Decreased Defective Impaired Absent Decreased neutrophil influx NA Absent NA Normal Late extravasation Normal Normal Reduced Normal

ICAM-1: intercellular adhesion molecule-1; NA: not applicable: NK: not known; VCAM-1: vascular cell adhesion molecule-1.

[18]. These mice made it possible to highlight two findings. Firstly, the frequency of rolling leukocytes in L-selectin-deficient mice treated with TNF was decreased significantly by the additional loss of ICAM-1 expression. Secondly, leukocyte rolling velocities were significantly greater in ICAM-/- mice than in wild-type mice. ICAM-1 influences selectin-mediated rolling of leukocytes, in addition to its role in firm adhesion and transmigration of leukocytes at the sites of inflammation [18]. Recent data highlight the fact that the up-regulation of ICAM-1 expression of LPS-induced ECs in vitro and in vivo is mediated by the p38 mitogen-activated protein kinase (MAPK) pathway at the level of gene transcription [19]. VCAM-1 mediates the adhesion of lymphocytes, monocytes, and eosinophils to activated endothelium. Its expression on ECs is induced by IL-1b, IL-4, TNF, and IFN-g. VCAM-1-deficient mice are not viable. Table 1 summarizes these data. Members of the selectin and Ig families function synergistically to mediate optimal leukocyte rolling and entry into tissues, which is essential for the generation of effective inflammatory response in vivo [20]. Future studies should investigate the Igs that are involved in the cell-to-cell adhesion, such as platelet EC adhesion molecule-1 (PECAM-1) or CD31, vascular endothelialcadherin (VE-cadherin), and CD146. PECAM-1 is involved in the process of neutrophil and monocyte diapedesis between ECs. When an anti-PECAM-1 is administered, light and electron microscopy provide evidence that leukocytes blocked in transmigration remain bound to the endothelial surface over the intercellular junction, thus clearly defining the role of PECAM-1 in diapedesis between ECs, rather than adherence to ECs [16]. VE-cadherin is an important determinant of vascular integrity in some organs. Leukocyte activation may lead to cleavage of the extracellular domain of VE-cadherin, which would increase in permeability in the areas of

leukocyte deposition [21]. CD146 is associated with actin cytoskeleton and is expressed at areas of cell-to-cell junction, which strongly suggests its involvement as a mediator of cell-to-cell interaction [22]. Figure 1 summarizes the interactions of leukocytes with endothelium. Nitric oxide and cell adhesion molecule interactions Septic shock is defined as sepsis with hypotension despite adequate fluid replacement resulting in impaired tissue perfusion and oxygen extraction [2]. Endotoxin or LPS binds to CD14 on the surface of cells (monocytes, macrophages, and some granulocytes), resulting in the release of proinflammatory cytokines. Binding of these cytokines to their specific receptors results in activation of protein kinase leading to phosphorylation of intracellular protein and activation of the nuclear transcription factor, nuclear factor kB (NF-kB). This activation leads to the proteolytic cleavage of its inhibitor, IkB, from NF-kB, which enables the activated NF-kB to enter the nucleus, where it binds to the promoter region of inducible nitric oxide synthase (iNOS) gene to activate transcription. Translation of iNOS mRNA is related to an enhanced formation of NO, which, in turn, may contribute to the pathophysiology of septic shock [23]. The denudation of endothelium leads to an abnormal vasodilatation by blunting the NO secretion. The endothelium reacts to hypoxia by inducing a sustained vasoconstriction through the release of endothelialderived contracting factors which act by scavenging NO [24]. NO is believed to modulate leukocyteEC interactions by acting as an endogenous anti-adhesive molecule. In vitro studies indicate that NO can attenuate EC adhesion molecule expression [25]. The results of a study investigating the iNOSselectin interactions indicated that the production of NO by iNOS plays a critical role in the expression of E- and P-selectin during

58

ADVANCES IN SEPSIS Vol 2 No 2 2002

ACTIVATION OF ADHESION MOLECULES IN SEPTIC SHOCK PATIENTS

Figure 1. Interactions of leukocytes with endothelium. A cascade of multiple molecular interactions controls the entry of leukocytes into tissue. Initial tethering of leukocytes to the endothelial cell surface is mediated by selectins. The leukocyte rolls along the blood vessel wall. The signaling of chemokines leads to activation of leukocyte integrins that bind to members of the immunoglobulin superfamily and mediate firm adhesion.
Adhesion of leukocytes l ICAM-1 and ICAM-2 l VCAM-1 l MAdCAM-1 Tethering Rolling Activation Firm-adhesion Migration Chemotaxis

L-selectin Integrin Integrin

P-selectin E-selectin

Transendothelial migration l PECAM-1 l ICAM-1 l VCAM-1

Chemokine Activation of ECs l Up-regulation of selectins l Synthesis of chemokines l Synthesis of PAF

ECs: endothelial cells; ICAM-1, -2: intercellular adhesion molecules-1 and -2; MAdCAM-1: mucosal addressin cell adhesion molecule-1; PAF: platelet activating factor; PECAM-1: platelet endothelial cell adhesion molecule-1; VCAM-1: vascular cell adhesion molecule-1.

polymicrobial sepsis in vivo [25]. A reduction in sepsis-induced E- and P-selectin expression on ECs was shown in iNOS/ mice compared with wild-type mice. This reduction was associated with decreased polymorphonuclear leukocyte accumulation in the gut, and suggests that NO plays an important role in modulating sepsis-induced EC adhesion molecule expression, and leukocyte sequestration [25]. Recent data provide evidence that a decrease in leukocyte rolling was related to a defect in the expression of adhesion molecules on endothelial surfaces mediated by iNOSderived NO [26]. The generation of NO in sepsis reduces neutrophil migration by inhibiting leukocyte rolling and their firm adhesion to the endothelium, in effect impairing the migration of leukocytes and, consequently, their fundamental role in host cell defense mechanisms. Hypoxia and cell adhesion molecule activation Oxygen deprivation leads to an inflammatory response. Because of their localization at the interface between blood and tissues, ECs are the first cells to undergo

damage when exposed to hypoxia. The activation of ECs leads to the expression of P-selectin. In experiments designed to establish the effect of hypoxia and hypoxiareoxygenation in vitro on human ECs, data have provided evidence that hypoxia alone is sufficient to induce P-selectin expression [27]. The inhibition of protein synthesis does not abrogate the hypoxia-induced expression of P-selectin on EC surfaces, proving that de novo protein synthesis was not required for this response [24]. Furthermore, protein synthesis of adhesion molecules and cytokines are required, as a study using bovine aortic endothelial monolayers exposed to hypoxia demonstrated [28]. In this model, hypoxia alone did not induce endothelial E-selectin expression. However, enhanced induction was observed when hypoxia was combined with either TNF or LPS. In addition, data has provided evidence that both anoxia/reoxygenation-induced E-selectin expression and neutrophilEC adhesion are mediated by both protein tyrosine kinase and protein kinase C, which signal rapid activation of NF-kB [29].

ADVANCES IN SEPSIS Vol 2 No 2 2002

59

MARC LEONE AND CLAUDE MARTIN

Table 2. Studies assessing adhesion molecules in septic shock patients.

Author [Ref] Newman et al. [31] Sessler et al. [32] Disease state Bacteremia Septic shock Systemic inflammation Septic shock Trauma versus sepsis Molecules sE-selectin sICAM-1 Findings Increased levels in septic shock patients Plasma levels: septic shock > systemic inflammation Increased levels in non-survivors Plasma levels: sepsis >trauma Increased levels in non-survivors Higher levels in serum of patients with documented sepsis Correlation with organ dysfunction Plasma levels correlated with the severity, but are not an accurate reflect of tissue expression Increased levels in non-survivors Identification of patients with risk of liver dysfunction Plasma levels: septic shock >sepsis >trauma Lack of correlation with skin biopsies

Boldt et al. [33]

sE-selectin sICAM-1 sVCAM-1 sE-selectin E-selectin ICAM-1 VCAM-1 sICAM-1 E-selectin P-selectin ICAM-1 VCAM-1

Cummings et al. [34] Turner et al. [35]

Sepsis and critical illness Mild and severe malaria

Weigand et al. [36] Leone et al. [37]

Septic shock Septic shock Severe sepsis Trauma

ICAM-1: intercellular adhesion molecule-1; s: soluble; VCAM-1: vascular cell adhesion molecule-1.

As we have seen, ICAM-1 and VCAM-1 play a crucial role for extravasation of white blood cells. To assess the influence of oxygen deprivation on basal- and cytokineinduced expression of both adhesion molecules, cells were incubated for up to 24 h at 21% and 0% O2 in the absence or presence of TNF [30]. Hypoxia led to a timedependent reduction of the cellular expression of ICAM-1 and VCAM-1, but the response of ECs to TNF was not significantly different in cells exposed to 0% O2 compared with the incubation at 21% O2. Subsequent increases in O2 levels (21%, 40%, or 95% O2) led to a marked increase of ICAM-1 and VCAM-1 cell surface and mRNA expression in ECs, which, after 16 h, amounted to approximately one-third to one-half of maximal TNF-induced expression. These data suggest that anoxia followed by reperfusion results in enhanced adhesivity of neutrophils. This situation can be paralleled with the state of the endothelium during sepsis. Cell adhesion molecules in septic shock patients Soluble cell adhesion molecules The concentrations of cell adhesion molecules in plasma have been measured in septic shock patients (Table 2) [3137]. A sustained increase of TNF, sE-selectin, sP-selectin, sICAM-1, and sVCAM-1 confirms that the marked activation of endothelium is associated with the more severe septic patients [3137]. Soluble forms of these endothelial adhesion molecules have been found to be increased in various inflammatory pathological

situations, and reflect the in vivo activation status of the endothelium. Increased levels of sICAM-1, sVCAM-1, and sE-selectin provide evidence that vascular endothelium is activated in septic shock patients, probably in response to high levels of TNF found in these patients. Interestingly, patients with bacteremia and hypotension showed markedly elevated E-selectin levels than those without hypotension, suggesting that the release of this adhesion molecule is sensitive to the degree of endothelial injury caused by sepsis [31]. In another study, higher levels of adhesion molecules found in septic shock patients have been compared with patients with severe sepsis. These data also reflected the higher degree of vascular injury in the more severe stages of sepsis [37]. In addition, E-selectin levels in patients with documented bacteremia are higher than in patients without microbiological documentation, suggesting pathogenendothelium interrelations [34]. Nonsurvivors show markedly increased plasma levels of all adhesion molecules [33]. In agreement with these results, a correlation was found between the plasma levels of adhesion molecules and the outcome of patients [38]. Little is known about the change of sP-selectin levels in sepsis. High circulating levels of this molecule have been reported in sepsis [36], with higher concentrations in patients with acute lung injury [39]. The levels of sP-selectin were more elevated in patients with septic shock than in patients with severe sepsis, again suggesting that the plasma level of this selectin is

60

ADVANCES IN SEPSIS Vol 2 No 2 2002

ACTIVATION OF ADHESION MOLECULES IN SEPTIC SHOCK PATIENTS

proportional to the degree of endothelial injury caused by sepsis. In contrast to sE-selectin, sP-selectin is not specific to the endothelium, so elevated levels of sP-selectin are, therefore, consistent with signs of both endothelium and platelet activation. These data show that a marked systemic activation of adhesion molecules occurs in septic shock patients, and this activation is related to the severity of the illness. Sustained endothelial activation may explain the increase in the soluble form of adhesion molecules that are observed in plasma. The molecules involved in the cell-tocell junction, such as PECAM-1, VE-cadherin, and CD146 have not been extensively studied in this clinical setting. In situ activation of adhesion molecules An animal study provided objective quantitative data showing that sICAM-1 does not accurately reflect the density of the membranous form of ICAM-1 in the vasculature [40]. To elucidate the difference between the soluble form and in situ density of cell adhesion molecules, various models were constructed. Different states of SIRS were compared in order to characterize the endothelium in septic shock. Levels of circulating soluble cell adhesion molecules were compared in septic shock and trauma patients, in whom a state of SIRS was observed [41]. Studies found higher levels of cell adhesion molecules in septic shock than in trauma patients [33,37,42]. This finding probably reflects a lower degree of endothelial activation in trauma patients compared with septic shock patients [33,37,42]. In order to correlate plasma levels and tissue expression of adhesion molecules, tissue biopsies were collected in the two groups of patients. Endothelial activation was assessed by documenting the expression of E-selectin, ICAM-1, and VCAM-1 in vivo in dermal vessels [37]. Surprisingly, similar expressions of sE-selectin, sICAM-1, and sVCAM-1 were observed in septic shock and in trauma patients, suggesting the absence of a correlation between plasma and tissue expression. A lack of correlation between adhesion molecule density on tissues and plasma levels of adhesion molecules was also reported in clinical diseases such as malaria [35], SIRS [43], and inflammatory bowel disease [44]. For instance, the levels of soluble forms of adhesion molecules in malaria are not an accurate reflection of EC expression of adhesion molecules in a particular vascular bed, and should not be related to the severity of disease [35]. Experimental data obtained in baboons showed differences in endothelial activation according to the vascular beds, with strong activation in the endothelium of the lung, liver, and kidneys, whereas it was weakest in

muscle [42]. This suggests that other tissues should be analyzed in order to have a better view of endothelial activation in sepsis. This could be explained by the phenotypic differences between activated endothelial phenotype in dermal vessels compared with other tissues, by the extraordinarily high levels of cytokines found in systemic sepsis [41], or by a delay in soluble adhesion molecule clearance in septic shock patients [35]. Therapeutic perspectives The potential therapeutic implications of these findings should be considered. Attempts to attenuate neutrophilmediated EC damage have been aimed at the blockade of essential adhesion molecule interactions. Selectin-derived peptides have been shown to inhibit neutrophil infiltration into sites of inflammation in a mouse peritonitis model [45]. In rats, anti-E-selectin antibodies inhibited neutrophil influx in glycogen-induced peritonitis, and attenuated neutrophil-mediate lung injury following immune complex deposition [46]. Anti-ICAM-1 antibodies have been shown to inhibit leukocyte infiltration and tissue injury in several models of lung inflammatory disease [47,48]. For instance, inhibition of the p38 MAPK signal pathway may be used as an approach to attenuate ICAM-1 production in the treatment of septic shock [19]. However, these therapies, limiting tissue injury in septic shock and reducing the progression of MOD syndromes, cannot be considered at the present time in humans [49]. Conclusion The higher levels of soluble forms of cell adhesion molecules in septic shock than in trauma patients indicate a greater systemic activation of the endothelium in sepsis and septic shock. However, the comparable endothelial activation, as evidenced by skin biopsies, suggests that caution is required in the interpretation of levels of soluble cell adhesion molecules in plasma a black box that does not necessarily reflect the in situ activation state of the endothelium. An understanding of the pathway of adhesion molecules involved in septic shock could lead to novel therapeutic approaches. References
1. Esmon CT, Fukudome K, Mather T et al. Inflammation, sepsis, and coagulation. Haematologica 1999;84:2549. 2. Parrillo JE. Pathogenetic mechanisms of septic shock. N Engl J Med 1993;328:14717. 3. Bevilacqua MP, Stengelin S, Gimbrone MA et al. Endothelial leukocyte adhesion molecule 1: An inducible receptor for neutrophils related to complement regulatory proteins and lectins. Science 1989;243:11605.

ADVANCES IN SEPSIS Vol 2 No 2 2002

61

MARC LEONE AND CLAUDE MARTIN

4. Bochner BS, Luscinskas FW, Gimbrone MA et al. Adhesion of humans basophils, eosinophils, and neutrophils to interleukin 1-activated human vascular endothelial cells: Contributions of endothelial cell adhesion molecules. J Exp Med 1991;173:15537. 5. Engelberts I, Samyo SK, Leeuwenberg JF et al. A role for ELAM-1 in the pathogenesis of MOF during septic shock. J Surg Res 1992;53:13644. 6. Johnston GI, Cook RG, McEver RP. Cloning of GMP-140, a granule membrane protein of platelets and endothelium: Sequence similarity to proteins involved in cell adhesion and inflammation. Cell 1989;56:103344. 7. Geng JG, Bevilacqua MP, Moore KL et al. Rapid neutrophil adhesion to activated endothelium mediated by GMP-140. Nature 1990;343:75760. 8. Yang J, Hirata T, Croce K et al. Targeted gene disruption demonstrates that P-selectin glycoprotein ligand 1 (PSGL-1) is required for P-selectin-mediated but not E-selectin-mediated neutrophil rolling and migration. J Exp Med 1999;190:176982. 9. Andre P, Hartwell D, Hrachovinova I et al. Pro-coagulant state resulting from high levels of soluble P-selectin in blood. Proc Natl Acad Sci USA 2000;97:1383540. 10. Eppihimer MJ, Russell J, Langley R et al. Role of tumor necrosis factor and interferon gamma in endotoxin-induced E-selectin expression. Shock 1999;11:937. 11. Vestweber D, Blanks JE. Mechanisms that regulate the function of the selectins and their ligands. Physiol Rev 1999;79:181213. 12. Ley K, Gaehtgens P, Fennie C et al. Lectin-like cell adhesion molecule 1 mediates leukocyte rolling in mesenteric venules in vivo. Blood 1991;77:25535. 13. Hickey MJ, Forster M, Mitchell D et al. L-selectin facilitates emigration and extravascular locomotion of leukocytes during acute inflammatory responses in vivo. J Immunol 2000;165:716470. 14. Homeister JW, Zhang M, Frenette PS et al. Overlapping functions of E- and P-selectin in neutrophil recruitment during acute inflammation. Blood 1998;92:234552. 15. Bullard DC, Kunkel EJ, Kubo H et al. Infectious susceptibility and severe deficiency of leukocyte rolling and recruitment in E-selectin and P-selectin double mutant mice. J Exp Med 1996;183:232936. 16. Cines DB, Pollak ES, Buck CA et al. Endothelial cells in physiology and in the pathophysiology of vascular disorders. Blood 1998;91:352761. 17. Sligh JE Jr, Ballantyne CM, Rich SS et al. Inflammatory and immune responses are impaired in mice deficient in intercellular adhesion molecule 1. Proc Natl Acad Sci USA 1993;90:852933. 18. Steeber DA, Campbell MA, Basit A et al. Optimal selectin-mediated rolling of leukocytes during inflammation in vivo requires intercellular adhesion molecule-1 expression. Proc Natl Acad Sci USA 1998;95:75627.

19. Yan W, Zhao K, Jiang Y et al. Role of p38 MAPK in ICAM-1 expression of vascular endothelial cells induced by lipopolysaccharide. Shock 2002;17:4338. 20. Steeber DA, Tang ML, Green NE et al. Leukocyte entry into sites of inflammation requires overlapping interactions between the L-selectin and ICAM-1 pathways. J Immunol 1999;163:217686. 21. Corada M, Mariotti M, Thurston G et al. Vascular endothelialcadherin is an important determinant of microvascular integrity in vivo. Proc Natl Acad Sci USA 1999;96:981520. 22. Bardin N, Anfosso F, Mass JM et al. Identification of CD146 as a component of the endothelial junction involved in the control of cellcell cohesion. Blood 2001;98:367784. 23. Thiemermann C. Nitric oxide and septic shock. Gen Pharmac 1997;29:15966. 24. Ten VS, Pinsky DJ. Endothelial response to hypoxia: Physiologic adaptation and pathologic dysfunction. Curr Opin Crit Care Med 2002;8:24250. 25. Lush CW, Cepinskas G, Sibbald WJ et al. Endothelial E- and P-selectin expression in iNOS-deficient mice exposed to polymicrobial sepsis. Am J Physiol Gastrointest Liver Physiol 2001;280:G2917. 26. Benjamim CF, Silva JS, Fortes ZB et al. Inhibition of leukocyte rolling by nitric oxide during sepsis leads to reduced migration of active microbicidal neutrophils. Infect Immun 2002;70:360210. 27. Closse C, Seigneur M, Renard M et al. Influence of hypoxia and hypoxia-reoxygenation on endothelial P-selectin expression. Thromb Res 1997;85:15964. 28. Zund G, Nelson DP, Neufeld EJ et al. Hypoxia enhances stimulusdependent induction of E-selectin on aortic endothelial cells. Proc Natl Acad Sci USA 1996;93:707580. 29. Kokura S, Rhoads CA, Wolf RE et al. NF kappa b signaling in posthypoxic endothelial cells: Relevance to E-selectin expression and neutrophil adhesion. J Vasc Res 2001;38:4758. 30. Willam C, Schindler R, Frei U et al. Increases in oxygen tension stimulate expression of ICAM-1 and VCAM-1 on human endothelial cells. Am J Physiol 1999;276:H204452. 31. Newman W, Beall LD, Carson CW et al. Soluble E-selectin is found in supernatants of activated endothelial cells and is elevated in the serum of patients with septic shock. J Immunol 1993;150:64454. 32. Sessler CN, Windsor AC, Schwartz M et al. Circulating ICAM-1 is increased in septic shock. Am J Respir Crit Care Med 1995;151:14207. 33. Boldt J, Mller M, Khun D et al. Circulating adhesion molecules in the critically ill: A comparison between trauma and sepsis patients. Intensive Care Med 1996;22:1228. 34. Cummings CJ, Sessler CN, Fisher BJ et al. Soluble E-selectin levels in sepsis and critical illness. Correlation with infection and hemodynamic dysfunction. Am J Respir Crit Care Med 1997;156:4317.

62

ADVANCES IN SEPSIS Vol 2 No 2 2002

ACTIVATION OF ADHESION MOLECULES IN SEPTIC SHOCK PATIENTS

35. Turner GD, Ly VC, Nguyen TH et al. Systemic endothelial activation occurs in both mild and severe malaria. Correlating dermal microvascular endothelial cell phenotype and soluble cell adhesion molecules with disease severity. Am J Path 1998;152:147787. 36. Weigand MA, Schmidt H, Pourmahmoud M et al. Circulating intercellular adhesion molecule-1 as an early predictor of hepatic failure in patients with septic shock. Crit Care Med 1999;27:265661. 37. Leone M, Boutire B, Camoin-Jau L et al. Systemic endothelial activation is greater in septic than in traumatic-hemorrhagic shock but does not correlate with endothelial activation in skin biopsies. Crit Care Med 2002;30:80814. 38. Kayal S, Jais JP, Aguini N et al. Elevated circulating E-selectin, intercellular adhesion molecule 1, and von Willebrand factor in patients with severe infection. Am J Respir Crit Care Med 1998;157:77684. 39. Sakamaki F, Ishizaka A, Handa M et al. Soluble form of P-selectin is elevated in acute lung injury. Am J Respir Crit Care Med 1995;151:18216. 40. Komatsu S, Flores S, Gerritsen ME et al. Differential up-regulation of circulating soluble and endothelial cell intercellular adhesion molecule-1 in mice. Am J Path 1997;151:20514. 41. Martin C, Boisson C, Haccoun M et al. Patterns of cytokine evolution (tumor necrosis factor-alpha and interleukin-6) after septic shock, hemorrhagic shock, and severe trauma. Crit Care Med 1997;25:18139.

42. Redl H, Dinges HP, Buurman WA et al. Expression of endothelial leukocyte adhesion molecule-1 in septic but not traumatic/hypovolemic shock in the baboon. Am J Pathol 1991;139:4616. 43. McGill SN, Ahmed NA, Hu F et al. Shedding of L-selectin as a mechanism for reduced polymorphonuclear neutrophil exudation in patients with systemic inflammatory response syndrome. Arch Surg 1996;131:11417. 44. Jones SC, Banks RE, Haidar A et al. Adhesion molecules in inflammatory bowel disease. Gut 1995;36:72430. 45. Briggs JB, Oda Y, Gilbert JH et al. Peptides inhibit selectin-mediated cell adhesion in vitro, and neutrophil influx into inflammatory sites in vivo. Glycobiology 1995;5:5838. 46. Mulligan MS, Varani J, Dame MK et al. Role of endothelialleukocyte adhesion molecule-1 (ELAM-1) in neutrophil-mediated lung injury in rats. J Clin Invest 1991;88:1396406. 47. Wegner CD, Gundel RH, Reilly P et al. Intracellular adhesion molecule-1 (ICAM-1) in the pathogenesis of asthma. Science 1990;247:4569. 48. Horgan MJ, Ge M, Gu J et al. Role of ICAM-1 in neutrophilmediated lung vascular injury after occlusion and reperfusion. Am J Physiol 1991;261:H157884. 49. Lin E, Calvano SE, Lowry SF. Selectin neutralization: Does it make biological sense? Crit Care Med 1999;27:20503.

ADVANCES IN SEPSIS Vol 2 No 2 2002

63