Neutrophil Adhesion Molecule Expression Research Articles

Listeria monocytogenes infection of cultured endothelial cells stimulates neutrophil adhesion and adhesion molecule expression.

Microbial infection of the endothelium with the resultant up-regulation of adhesion molecule expression and stimulated leukocyte adhesion to endothelial cells can promote an inflammatory response. Previous work demonstrated that Listeria monocytogenes can replicate within cultured endothelial cells; thus, we tested whether L. monocytogenes infection of HUVEC stimulated an inflammatory phenotype on these cells. Infection with 10(4) CFU of bacteria increased neutrophil adhesion to HUVEC 40-fold and up-regulated E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 expression. Approximately 80% of neutrophil adhesion to infected HUVEC was blocked by anti-E-selectin mAb, 35% was blocked by anti-CD18 mAb, and anti-vascular cell adhesion molecule-1 mAb was without effect. Microscopy of infected HUVEC monolayers showed that neutrophils bound to infected and uninfected cells and that infected and uninfected HUVEC expressed E-selectin. Interestingly, uninfected HUVEC that bound neutrophils or expressed E-selectin typically were adjacent to infected cells. However, infected monolayers did not produce soluble factors that stimulated E-selectin expression on uninfected cells. Nuclear translocation of the p65 subunit of the transcription factor NF-kappaB accompanied the HUVEC response, and hemolysin secretion appeared critical for stimulating HUVEC. These studies show that L. monocytogenes infection stimulates up-regulation of adhesion molecule expression on endothelial cells, resulting in neutrophil adhesion to them. This response includes induction of an inflammatory phenotype on uninfected cells and may be triggered by listeriolysin O-mediated activation of host response mechanisms. Additionally, cell-to-cell spread of L. monocytogenes throughout the monolayer, without stimulating secondarily infected endothelial cells for neutrophil adhesion, is a possible means of immune avoidance.

Coronary Angioplasty Results in Leukocyte and Platelet Activation With Adhesion Molecule Expression: Evidence of Inflammatory Responses in Coronary Angioplasty

Objectives. This study sought to characterize leukocyte and platelet activation and adhesion molecule expression after coronary angioplasty.Background. Coronary angioplasty can be regarded as a clinical model of postischemic inflammation because this intervention leads to the release of inflammatory mediators as a result of plaque rupture and endothelial injury.Methods. In 13 patients with stable angina (mean [±SEM] age 56.0 ± 2.4 years, range 44 to 79), blood samples were drawn from the aorta and coronary sinus immediately before and immediately and 15 min after coronary angioplasty. Subsequently, leukocyte and platelet functions were determined. Eleven control patients (57.5 ± 2.3 years, range 52 to 78) underwent coronary arteriography.Results. Coronary arteriography and angioplasty showed no difference in number of leukocytes between the coronary sinus and the aorta. However, 15 min after coronary angioplasty, there was an increase in neutrophil CD18 and CD11b, monocyte CD14 and platelet glycoprotein IIb/IIIa expression and a decrease in neutrophil L-selectin expression (189 ± 25%, 163 ± 27%, 158 ± 35%, 141 ± 22% and 31 ± 10%, respectively, p < 0.01). In the control subjects, no change in adhesion molecule expression occurred. Superoxide production and aggregation in ex vivo-stimulated neutrophils collected from the coronary sinus 15 min after coronary angioplasty was significantly decreased compared with that after coronary arteriography (54 ± 12% vs. 106 ± 30% and 58 ± 11% vs. 102 ± 29%, respectively, p < 0.01). The reduced responses to phorbol ester stimulation may be explained by previous in vivo activation of neutrophils during coronary angioplasty.Conclusions. Coronary angioplasty increases neutrophil, monocyte and platelet adhesion molecule expression and induces a significant decrease in ex vivo-stimulated neutrophil superoxide generation and aggregation. These findings suggest that coronary angioplasty triggers cellular activation with an inflammatory response that could contribute to restenosis.(J Am Coll Cardiol 1997;29:1276–83)

Neutrophil adhesion molecule expression and response to stimulation with bacterial wall products in humans is unaffected by parenteral nutrition.

1. Total parenteral nutrition is associated with a high incidence of septic complications. This may be partly due to neutrophil dysfunction induced by the parenteral nutrition. 2. Neutrophil adhesion molecule expression and the expression of CD11b in response to stimulation with formylmethionyl-leucyl-phenylalanine and lipopolysaccharide were determined before and after 24 h of lipid-containing parenteral nutrition. Eighteen adult patients referred for parenteral nutrition were studied. 3. There was no change in the expression of neutrophil L-selectin (CD62L), CD11a, CD11b, CD11c or CD15. Neutrophil response to stimulation with formylmethionyl-leucyl-phenylalanine and lipopolysaccharide as determined by CD11b expression was unaffected by parenteral nutrition. 4. This study has shown no evidence of parenteral nutrition-induced neutrophil dysfunction.

Haemostatic changes in the pulmonary blood during cardiopulmonary bypass.

Pulmonary injury may result from the use of cardiopulmonary bypass (CPB). We investigated changes in the haemostatic system in the pulmonary vein during CPB compared with blood that circulated through the bypass circuit. Paired samples were taken from the pulmonary vein and central venous pressure (CVP) line during the peri-operative period from ten patients. Plasma levels of factor VII (P < 0.001), prekallikrein (P < 0.05), antithrombin III (P < 0.001) and heparin cofactor II (P < 0.005) were decreased in the pulmonary vein after 20 min of bypass compared with pre-operative levels. In the pulmonary vein there was a significant increase in neutrophil expressed CD11b (P < 0.001), neutrophil elastase: alpha 1-antitrypsin complexes (P < 0.001), endothelin-1(P < 0.001) and thrombin-antithrombin complexes (P < 0.001) by the end of bypass compared with pre-operative levels. There was no significant change in monocyte expressed CD11b, factor XII or C1-esterase inhibitor in the pulmonary vein for the study period. None of these variables were significantly different in the pulmonary vein compared with CVP line. In the pulmonary vein plasma levels of activated factor VII decreased following heparin administration (P < 0.001) in the majority of patients which was coincidental to an increase (P < 0.001) in tissue factor pathway inhibitor (TFPI). This increase in TFPI was significantly higher in the pulmonary vein compared with CVP line (P < 0.05) There was a decrease in neutrophil count by 20 min on CPB in both the pulmonary vein and CVP line (P < 0.001) and this did not return to pre-operative levels in the pulmonary vein. Soluble thrombomodulin levels decreased by 20 min on CPB in the CVP line (P < 0.05) but tended to increase in the pulmonary vein, although this was not statistically significant. In conclusion we found evidence of thrombin generation and possible endothelial damage together with increased neutrophil activation and adhesion molecule expression in the pulmonary vein during CPB which may play an important role in the development of post-CPB pulmonary injury.

Obstructive jaundice causes reduced expression of polymorphonuclear leucocyte adhesion molecules and a depressed response to bacterial wall products in vitro.

Obstructive jaundice is associated with an increased incidence of infection and endotoxaemia, which may result from impaired host immunity. Neutrophil adhesion to vascular endothelium is a key part of the inflammatory response. To investigate neutrophil adhesion molecule expression and activation in obstructive jaundice. Nine adult patients with obstructive jaundice and 11 control subjects. The expression of the neutrophil adhesion receptors L-selectin, CD11a, CD11b, CD11c, and CD15 was determined using flow cytometry. CD11b expression in response to stimulation with fMLP and endotoxin was measured. The basal expression of L-selectin, CD11a, and CD15 was significantly decreased in jaundiced patients (p < 0.05) and the expression of CD11b in response to stimulation with fMLP and endotoxin was significantly impaired in the jaundiced group. Endotoxin stimulation without plasma did not reverse the impaired response showing that it is not caused by endotoxin inactivation by plasma proteins. Neutrophils from patients with obstructive jaundice show decreased adhesion receptor expression and an impaired response to stimulation with bacterial products. This cellular dysfunction may be responsible for the high incidence of septic complications in these patients.

The influence of capsulation and lipooligosaccharide structure on neutrophil adhesion molecule expression and endothelial injury by Neisseria meningitidis.

Host inflammatory response to meningococcal infection is believed to be a major determinant of disease severity. Isogenic mutants of Neisseria meningitidis serogroup B1940, which differ in expression of capsular polysaccharide and lipooligosaccharide (LOS), were used to examine host responses in a whole blood model of bacteremia and a model of endothelial injury. The parent organism caused significantly less neutrophil shedding of the adhesion molecule, L-selectin, than the three mutant organisms (P < .01) and was most resistant to the bactericidal activity of whole blood. Despite marked differences in bacterial adhesion to endothelial cells (P < .05), no damage was induced by organisms alone. Endothelial injury was observed when neutrophils were incubated with adherent, capsule-deficient organisms (P < .05). The degree of endothelial damage was related to the number of neutrophils adherent to the endothelium. Thus, bacterial capsulation and LOS structure can influence neutrophil activation and endothelial injury and, as such, may be important in the pathogenesis of meningococcal sepsis.

Hypothermia during cardiopulmonary bypass delays but does not prevent neutrophil-endothelial cell adhesion. A clinical study.

An accurate evaluation of warm heart surgery cannot be limited to the assessment of the myocardial effects of warm blood cardioplegia but should also address the effects of systemic normothermia on the inflammatory response to cardiopulmonary bypass. A major component of this response is the endothelial adhesion of neutrophils, because it is linked to the release of cytotoxic compounds. This study was designed (1) to characterize the bypass-induced changes in the expression of neutrophil adhesion molecules (L-selectin and beta 2-integrins) and (2) to assess the influence of bypass temperature on these changes. Twenty case-matched patients undergoing open-heart procedures were divided into two equal groups according to the core temperature during cardiopulmonary bypass: warm (33.4 +/- 0.3 degrees C) or cold (27.1 +/- 0.4 degrees C, P < .0001 versus warm). Arterial blood samples were collected before, during, and 30 minutes after bypass and processed for the expression of L-selectin and beta 2-integrins (CD11a, CD11b, and CD11c) with flow cytometry. Warm bypass was associated with an early and sustained upregulation of CD11b. In contrast, hypothermia resulted in a strikingly less pronounced CD11b upregulation during bypass. However, CD11b expression sharply increased thereafter so that 30 minutes after bypass, it was no longer significantly different between the two groups. Changes in CD11c expression grossly paralleled those described for CD11b. Neither CD11a nor L-selectin changed significantly from baseline values in either group. Clinical cardiopulmonary bypass is associated with a marked upregulation of the neutrophil CD11b and CD11c integrins. Hypothermia delays but does not prevent the increased expression of these adhesion molecules, which could consequently represent logical targets for interventions designed to blunt the neutrophil-mediated component of bypass-induced inflammatory tissue damage.

Neutrophil activation and adhesion molecule expression in a canine model of open heart surgery with cardiopulmonary bypass

Neutrophil activation and adhesion molecule expression in a canine model of open heart surgery with cardiopulmonary bypass

Neutrophil activation and adhesion molecule expression in a canine model of open heart surgery with cardiopulmonary bypass

The aim was to determine whether, in a canine model, changes in surface expression of the neutrophil adhesion molecules CD11b/CD18 and L-selectin during and after open heart surgery with cardiopulmonary bypass can be used to identify subjects at risk for postoperative pulmonary dysfunction. Adult mixed breed dogs underwent cardiopulmonary bypass and were compared to "sham bypass" controls. Flow cytometry was performed on blood from the two groups of dogs and changes in CD11b/CD18 adhesion molecules and L-selectin were investigated. Flow cytometry on blood from bypass dogs showed increased CD18 expression during and after cardiopulmonary bypass and a reciprocal decrease in L-selectin expression. Sham animals showed no significant change. In the bypass animals, changes in adhesion molecule expression were not evenly distributed across the population of circulating neutrophils; however, they were indicative of a percentage of activated cells. There was a significant negative linear relationship between the percentage of activated cells and arterial oxygenation 3 h after bypass (r = -0.80, P < 0.001). From this analysis, 11 animals were identified as "high" responders and seven as "low" responders, with different patterns of cellular activation and oxygenation during and after bypass. High responders had an average of 40(SEM 5)% activated cells during bypass with a persistently raised percentage of activated cells [38(3)%] 3 h later, whereas low responders had only 22(6)% activated cells during bypass and 11(2)% activated cells 3 h after bypass. High responder animals had a marked and continued deterioration in PO2 after bypass [to 25(6)% of baseline 3 h after bypass] whereas low responder animals showed recovery of oxygenation after the first hour postbypass and improved to 80(8)% of baseline at 3 h. Changes in adhesion molecule expression serve as a marker of neutrophil activation during cardiopulmonary bypass. The percentage of activated neutrophils in the circulation within 3 h after cardiopulmonary bypass may be predictive of an ongoing inflammatory process that is linked to pulmonary dysfunction.

C5a- and Tumor Necrosis Factor-α-Induced Leukocytosis Occurs Independently of β2 Integrins and L-Selectin: Differential Effects on Neutrophil Adhesion Molecule Expression In Vivo

We previously demonstrated in rabbits that various neutrophil chemotactic factors share an ability to induce recruitment of polymorphonuclear leukocytes (PMN) from bone marrow when administered intravenously (Jagels and Hugli, J Immunol 148:1119, 1992). In the study reported here, we investigated the effects of chemotactic factors on the expression of beta 2 integrins and L-selectin in vivo and the roles of these adhesionmolecules in the recruitment process. Leukocytosis was induced by infusion of either C5a (5 micrograms/kg), N-formyl-methionyl-leucyl-phenylalanine (f-MLP; 2.5 micrograms/kg), or tumor necrosis factor alpha (TNF-alpha; 100 ng/kg). C5a increased the expression of CD18 (the common subunit of beta 2 integrins) on PMN by nearly twofold and decreased levels of L-selectin by 50% within 15 minutes after administration. Levels of beta 2 integrins returned to baseline 2 to 3 hours after induction of leukocytosis. L-selectin remained depressed for more than 5 hours, demonstrating that shedding was induced in the recruited bone marrow leukocytes as well as in circulating PMN. In contrast to the response to C5a, TNF-alpha did not cause upregulation of CD18 or shedding of L-selectin. Levels of L-selectin were consistently increased 60 minutes after administration of TNF-alpha, coinciding with a rapid rise in the number of band-form PMN in the circulation. Intact IgG and F(ab)2 forms of the anti-CD18 monoclonal antibody IB4 or the anti-L-selectin antibody DREG-200 were administered intravenously 15 minutes before induction of leukocytosis by the chemotactic factors. Neither IB4 nor its F(ab)2 fragments blocked leukocytosis induced by C5a, f-MLP, or TNF-alpha. DREG-200 also did not block leukocytosis induced by f-MLP, C5a, or TNF-alpha. These results suggest that leukocyte emigration from the bone marrow into the circulation proceeds through interactions distinct from those involved in neutrophil chemotaxis and diapedesis. Shedding of L-selectin from C5a-recruited bone marrow leukocytes demonstrates activation of these cells in the recruitment process and may reflect a potential mechanism for their release. The dissimilar effects of C5a and TNF-alpha on expression of adhesion molecules may result from distinct stimulatory pathways and suggests differential activation states for cellular recruitment by these inflammatory factors.

Induction of neutrophil adhesion molecule (CD18) expression and polarization by rectal dialysates from patients with colitis and Crohn's disease

Induction of neutrophil adhesion molecule (CD18) expression and polarization by rectal dialysates from patients with colitis and Crohn's disease

Inhibition of neutrophil adhesion during cardiopulmonary bypass

Blood contact with synthetic surfaces during cardiopulmonary bypass (CPB) causes a diffuse inflammatory reaction that includes neutrophil activation. The purpose of this study was to determine if inhibition of neutrophil adhesion with a new antiinflammatory agent NPC 15669 ( N-(9H-(2,7-dimethylfluorenyl-9-methoxy)-carbonyl)- l-leucine) could reduce pulmonary injury in a porcine model of CPB. NPC 15669 blocks adherence of activated neutrophils by inhibiting upregulation of the Mac-1 ( CD11b / CD18 ) adhesion molecule. Sixteen piglets underwent 2 hours of hypothermic CPB followed by 2 hours of observation; 8 received NPC 15669 (10 mg/kg intravenous bolus followed by 6 mg · kg −1 · h −1 intravenous infusion) and 8 received equal volumes of vehicle. After 90 minutes of CPB, expression of neutrophil adhesion molecule subunit CD18 increased 118% in control piglets but only 36% in piglets treated with NPC 15669 ( p < 0.01). Although neutropenia developed in all animals during CPB, lung tissue myeloperoxidase content was significantly lower in treated than in control animals 2 hours after CPB (94.9 ± 10.4 versus 46.9 ± 5.5 μmol · 10 mg −1 · min −1; p < 0.002). Free radical-mediated lipid peroxidation (quantitated by spectrophotometric assay of plasma conjugated dienes) was significantly reduced by treatment with NPC 15669 during and after CPB. Pulmonary function was better in MPC 15669-treated animals: 2 hours after CPB, pulmonary vascular resistance increased 477% in control piglets but only 140% in piglets receiving NPC 15669 ( p < 0.03); arterial oxygen tension was significantly greater in piglets receiving NPC 15669 (428 ± 33 mm Hg) than in controls (141 ± 46; p < 0.0001. Histologic examination revealed neutrophil sequestration and interstitial and intraalveolar edema in control animals, but virtually normal lung architecture in animals that received NPC 15669. These results demonstrate that NPC 15669 reduced neutrophil adhesion molecule expression, pulmonary leukocyte sequestration, and free radical generation during CPB with a corresponding reduction in lung injury. These findings suggest that neutrophils are important mediators of CPB-associated pulmonary damage and that inhibition of neutrophil-endothelial adhesion is a promising new modality for reducing organ injury associated with CPB.

Humoral and cellular activation in a simulated extracorporeal circuit

Endothelial injury consequent upon widespread humoral and cellular activation is probably a major contributor to the phenomenon of cardiopulmonaty bypassinduced organ dysfunction. This article reviews some of the mechanisms by which complement and neutrophit activation and interleukin-8 may be involved in this inflammatory response. In a model consisting of a simulated extracorporeal circulation we were able to demonstrate complement activation, profound and specific changes in neutrophil adhesion molecule expression, and interleukin-8 generation. The importance of these changes and their potential interactions are discussed.

Eosinophil major basic protein enhances the expression of neutrophil CR3 and p150,95

Background: We examined the effect of major basic protein (MBP) stimulation on the expression of the neutrophilβ 2-integrins: LFA-1 (CD11a/CD18), CR3 (CD11b/CD18), and p150,95 (CD11c/CD18). Methods: Incubation of neutrophils with 0.75 to 3.0 μmol/L MBP for 30 minutes at 37 °C resulted in concentration-dependent increases in CR3 expression as detected by staining with the CD11b-specific monoclonal antibody, Leu-15. Results: The expression of CR3 was significantly (p < 0.001) higher when neutrophils were stimulated with 1.5 μmol/L (53% ± 9% increase) or 3.0 μmol/L (100% ± 17% increase) MBP as compared with unstimulated neutrophils. The kinetics of MBP-stimulated CR3 expression were rapid and were similar to those of 100 nmol/L N-formyl-methionyl-leucyl-phenylalanine-stimulated enhancement of CR3 expression. Incubation of neutrophils with reduced and alkylated MBP resulted in significantly lower (p < 0.05) increases in CR3 expression as compared with stimulation with native MBP. In addition, neither eosinophil cationic protein nor eosinophil-derived neurotoxin altered neutrophil CR3 expression. MBP stimulated minimal increases in LFA-1 expression. However, staining with the monoclonal antibody Leu-M5 (anti-CD11c) revealed that MBP stimulated significant increases in the expression of p150,95. Conclusions: These results indicate that MBP stimulates the increased expression of neutrophil adhesion molecules, which in turn may enhance the inflammatory role of neutrophils in the late-phase events of allergic diseases.

Neutrophil adhesion molecule expression during cardiopulmonary bypass with bubble and membrane oxygenators

The neutrophil-mediated tissue injury associated with cardiopulmonary bypass (CPB) is thought to require the interaction of specific neutrophil and endothelial adhesion molecules. In this study, the effects of CPB on the expression of neutrophil CD11b and CD18 (the components of the Mac-1 adhesion molecule) were examined; the effects of membrane versus bubble oxygenators on the expression of neutrophil CD11b and CD18 were compared; and the plasma levels of the intercellular adhesion molecule-1 (cICAM-1), an inducible endothelial adhesion molecule, were measured. In addition, the time courses of complement activation and neutrophil granule release were measured to determine their temporal relationship to the expression of the neutrophil adhesion molecule. Fifteen adult patients underwent procedures requiring cardiopulmonary bypass; hollow-fiber membrane oxygenators were used in 8 (group M) and bubble oxygenators were used in 7 (group B). Blood samples were drawn before, during, and after CPB for determination of the expression of neutrophil CD11b and CD18 (immunofluorescent flow cytometry), and the plasma cICAM-1, elastase, lactoferrin (enzyme-linked immunoabsorbent assay), and plasma C3a (radioimmunoassay) levels. CPB caused an immediate and sustained increase in the neutrophil CD11b and CD18 expression in both groups; after 60 minutes of CPB, CD11b expression had increased by 116.9% ± 19.1% in group B and by 79.3% ± 8.5% in group M ( p = 0.78). Over the same period, CD18 expression increased by 97.2% ± 17.9% in group B and by 72.4% ± 16.8% in group M ( p = 0.67). Increased neutrophil adhesion molecule expression was accompanied by elevations in the plasma elastase, lactoferrin, and C3a levels; there were no differences between oxygenator groups for any of these parameters. Before CPB, the plasma cICAM-1 levels were similar in the two groups (group B, 175.6 ± 25.6 ng/ml; group M, 200.2 ± 30 ng/ml). Twenty-four hours after CPB, the plasma cICAM-1 level increased to 289.1 ± 31.1 ng/ml in group B patients ( p < 0.004 versus the baseline) but did not increase in group M patients (220.5 ± 20.6 ng/ml). These results demonstrate that (1) CPB causes upregulation of the neutrophil adhesion molecule subunits CD11b and CD18; (2) the bubble and membrane oxygenators used in this study cause similar degrees of neutrophil adhesion molecule upregulation, neutrophil degranulation, and complement activation; and (3) use of bubble, but not membrane, oxygenators results in increased plasma cICAM-1 levels.

Effects of Human Neutrophil Elastase (HNE) on Neutrophil Function In Vitro and in Inflamed Microvessels

The primary objective of this study was to test the hypothesis that human neutrophil elastase (HNE) affects neutrophil infiltration (adhesion and emigration) into inflamed vessels. To determine whether HNE contributes to neutrophil adhesion in vivo, intravital microscopy was used to study neutrophil-endothelial cell interactions in single inflamed postcapillary venules. Superfusion of platelet-activating factor (PAF) (100 nmol/L) onto the mesentery caused an increase in neutrophil-neutrophil interactions, neutrophil adhesion to postcapillary venules, and cellular emigration out of the vasculature. Both L658 758 (an elastase-specific inhibitor), and Eglin C (an elastase and cathepsin G inhibitor) significantly attenuated all of these parameters in vivo. To further characterize the mechanism(s) involved, various in vitro parameters were assessed. HNE, but not trypsin, caused a dose-dependent (0.01 to 1.0 microgram/mL) increase in the expression of the beta subunit (CD18) of the CD11/CD18 adhesive glycoprotein complex on neutrophils. An HNE-dependent increase in CD11b expression was also observed; however, HNE did not affect the expression of other neutrophil adhesion molecules (L-selectin), superoxide production, or degranulation. PAF-enhanced CD18 expression on neutrophils and neutrophil migration were both abolished by L658 758 but PAF-induced neutrophil adhesion to endothelial monolayers was not affected by the antiproteinase. The in vitro data suggest that the antiproteinases do not directly prevent neutrophil adhesion in vivo but may be important in other CD18-dependent events such as neutrophil-neutrophil interaction or neutrophil infiltration (chemotaxis). These results translate into an important, rate-limiting role for elastase in the process of leukocyte infiltration and accumulation in inflamed microvessels.

Effects of human neutrophil elastase (HNE) on neutrophil function in vitro and in inflamed microvessels

The primary objective of this study was to test the hypothesis that human neutrophil elastase (HNE) affects neutrophil infiltration (adhesion and emigration) into inflamed vessels. To determine whether HNE contributes to neutrophil adhesion in vivo, intravital microscopy was used to study neutrophil-endothelial cell interactions in single inflamed postcapillary venules. Superfusion of platelet-activating factor (PAF) (100 nmol/L) onto the mesentery caused an increase in neutrophil-neutrophil interactions, neutrophil adhesion to postcapillary venules, and cellular emigration out of the vasculature. Both L658 758 (an elastase-specific inhibitor), and Eglin C (an elastase and cathepsin G inhibitor) significantly attenuated all of these parameters in vivo. To further characterize the mechanism(s) involved, various in vitro parameters were assessed. HNE, but not trypsin, caused a dose-dependent (0.01 to 1.0 microgram/mL) increase in the expression of the beta subunit (CD18) of the CD11/CD18 adhesive glycoprotein complex on neutrophils. An HNE-dependent increase in CD11b expression was also observed; however, HNE did not affect the expression of other neutrophil adhesion molecules (L-selectin), superoxide production, or degranulation. PAF-enhanced CD18 expression on neutrophils and neutrophil migration were both abolished by L658 758 but PAF-induced neutrophil adhesion to endothelial monolayers was not affected by the antiproteinase. The in vitro data suggest that the antiproteinases do not directly prevent neutrophil adhesion in vivo but may be important in other CD18-dependent events such as neutrophil- neutrophil interaction or neutrophil infiltration (chemotaxis). These results translate into an important, rate-limiting role for elastase in the process of leukocyte infiltration and accumulation in inflamed microvessels.

Heparin-coated bypass circuits reduce pulmonary injury

Heparin coating of the extracorporeal circuit not only reduces heparin requirements during cardiac operations but also may reduce organ injury associated with cardiopulmonary bypass (CPB). To examine this possibility, pulmonary injury and neutrophil adhesion molecule expression after CPB were studied in pigs undergoing CPB with a standard extracorporeal circuit (group S, n = 6) or a heparin-coated CPB circuit (Carmeda BioActive Surface) (group HC, n = 6). Pigs received heparin sodium (300 U/kg intravenously) and then underwent 90 minutes of hypothermie (28 °C) CPB using membrane oxygenators, followed by 2 hours of observation. Blood samples were obtained for determination of neutrophil number and expression of the neutrophil adhesion molecule subunit CD18 (by immunofluorescence flow cytometry). The CPB-associated injury was less in group HC. Two hours after CPB, the arterial oxygen tension group was higher in group HC (597.2 ± 31.2 versus 220.5 ± 42.3 mm Hg; p < 0.0001), the pulmonary vascular resistance was lower in these animals (408.6 ± 69.4 versus 1,159.8 ± 202.4 dyne · s · cm −5; p = 0.02), and the static compliance was higher in group HC (66.4 ± 5.4 versus 39.8 ± 5.8 mL/mm Hg; p = 0.004). After 60 minutes of CPB, both groups had similar increases in expression of the neutrophil adhesion molecule subunit CD18 (29.4% ± 19.5% versus 26.0% ± 24.4%, group S and group HC, respectively) and similar decreases in neutrophil counts (6,056 ± 1,285 to 2,453 ± 979 cells/μL versus 6,010 ± 1,748 to 3,197 ± 1,225 cells/μL, group S and group HC, respectively). This study demonstrates that heparin coating improves pulmonary function after CPB and that this effect is not mediated by altered neutrophil kinetics or adhesion molecule expression.

Increased expression of neutrophil and monocyte adhesion molecules in unstable coronary artery disease.

A rapid increase in leukocyte adhesion to endothelial cells is one of the first events in the acute inflammatory response and in the pathogenesis of vascular diseases. A subgroup of cell surface glycoproteins (the CD11/CD18 complex) play a major role in the leukocyte adhesion process; in particular, the CD11b/CD18 receptor can be upregulated severalfold in response to chemotactic factors. The purpose of this study was to assess whether upmodulation of granulocyte and monocyte CD11b/CD18 receptors takes place during the passage of blood through the coronary tree of patients with clinical manifestations of ischemic heart disease. Thirty-nine patients who underwent diagnostic coronary arteriography were studied. Group 1 (15 patients) had a clinical diagnosis of unstable angina, group 2 (14 patients) had stable exertional angina, and group 3 (10 patients) had atypical chest pain. Simultaneous sampling from the coronary sinus and aorta was obtained before coronary arteriography. Cell surface receptors were detected by direct immunofluorescence evaluated by flow cytofluorimetry using monoclonal antibodies tagged with fluorescent markers. Leukocytes were stained in unseparated blood to avoid in vitro manipulation that could activate phagocytes. Group 1 and 2 patients had significant coronary artery disease (> 50% coronary narrowing in at least one major coronary vessel), whereas group 3 patients had normal coronary arteries. In group 1, granulocytes and monocytes showed a significantly higher expression of the CD11b/CD18 adhesion receptor in the coronary sinus than in the aorta (both P < .01), whereas no difference in CD11b/CD18 expression was seen in groups 2 and 3. Patients with unstable angina have an increased expression of granulocyte and monocyte CD11b/CD18 adhesion receptors, indicating that an inflammatory reaction takes place within their coronary tree. Activation of these leukocytes may induce coronary vasoconstriction, favor thrombotic processes, and further activate platelets, thus having potential implications on the pathogenesis of unstable coronary artery disease.

5 Neutrophil activation and adhesion molecule expression in a cannie model of cardiopulmonary bypass (CPB)

5 Neutrophil activation and adhesion molecule expression in a cannie model of cardiopulmonary bypass (CPB)