Granulocyte Elastase Activity Research Articles

Relationship of changes in interleukin-8 levels and granulocyte elastase activity in gingival crevicular fluid to subgingival periodontopathogens following non-surgical periodontal therapy in subjects with chronic periodontitis.

To determine the effect of scaling and root planing (SRP) on the interrelations of subgingival periodontopathogens and both interleukin-8 (IL-8) and granulocyte elastase activity in gingival crevicular fluid (GCF), and to assess their relations to the short-term treatment response in management of chronic periodontitis. GCF and subgingival plaque were collected from 16 subjects with untreated chronic periodontitis at baseline and 4 weeks after SRP. IL-8 levels were determined by ELISA. Granulocyte elastase activity was analyzed with a specific substrate, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA) was calculated. 5 DNA-probes were used to detect the presence of A. actinomycetemcomitans (A. a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.), with a sensitivity = 103 cells/paper point. IL-8 and MR-EA levels in GCF decreased significantly after SRP (p < 0.001) with a corresponding reduction of total count of the species. Of the sites with probing depth (PD) >/= 5.0 mm and co-infection by B.f., P.g., P.i. & T.d. at baseline, the sites without persistent co-infection of these species after SRP exhibited a significant reduction of IL-8 levels (p < 0.02), MR-EA levels (p < 0.02) and PD (p < 0.01). No such change was found in the sites where such a co-infection persisted. Moreover, reduction of IL-8 levels in those pocket sites was accompanied by a concomitant reduction of MR-EA (p < 0.02) and PD (p < 0.01), while no significant change in MR-EA levels and PD was noted in those pocket sites that exhibited an increase of IL-8 levels after SRP. At baseline, the former group of sites showed significantly higher IL-8 levels than the latter group of sites (p < 0.02). IL-8-related granulocyte elastase activity was related to the change in infection patterns of the target periodontopathogens following scaling and root planing. Varying initial IL-8 levels in GCF and a corresponding shifting change of granulocyte elastase activity in GCF may characterize the different short-term treatment responses.

Granulocyte elastase activity and PGE2 levels in gingival crevicular fluid in relation to the presence of subgingival periodontopathogens in subjects with untreated adult periodontitis.

This study aimed to determine the association between the levels of granulocyte elastase and prostaglandin E2 (PGE2) in GCE and the concomitant presence of periodontopathogens in untreated adult periodontitis (AP). GCF and subgingival plaque were sampled by paper strips and paper points respectively, from various periodontal sites in 16 AP subjects. Granulocyte elastase activity in GCF was analyzed with a low molecular weight substrate specific for granulocyte elastase, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA, mAbs/min/site) was calculated. PGE2 levels in GCF were determined by radioimmunoassay. 5 species-specific DNA probes were used to detect the presence of A. actinomyceterncomitans (A.a., ATCC 43718), B. forsythus (B.f, ATCC 43037), P. gingivalis (P.g., ATCC 33277), P. intermedia (P.i., ATCC 33563), and T. denticola (T.d., ATCC 35405), with a sensitivity of 10(3) cells/paper point. No A.a. was detectable from all sites sampled. The predominant combination of species detected was B.f., P.g., P.i. & T.d. and it was significantly higher at periodontitis sites (68%) than at healthy (7%) or gingivitis sites (29%) (p<0.05). Overall, MR-EA values were strongly correlated with PGE2 levels (r=0.655, p<0.001), especially at these periodontitis sites co-infected by B.f., P.g., P.i. & T.d. (r=0.722, p<0.001). The periodontitis sites co-infected by the 4 species were observable from 15 subjects. These sites were sub-grouped into 8 subjects with a high MR-EA and 7 subjects with a low MR-EA. The PGE2 levels in the high MR-EA group were significantly higher than in the low MR-EA group (p<0.05). No significant differences in clinical or bacterial data were found between the two groups. While within the high MR-EA group, similar results were found between the paired periodontitis sites in each subject with highest and lowest MR-EA values. This study shows that the local host response to bacterial challenge in untreated periodontal pockets is diverse in terms of the intensity of inflammatory response measured by granulocyte elastase and PGE2 levels in GCE A more thorough evaluation of the risk for active periodontal disease may involve the combined approaches to the test of the dynamic bacteria-host relations.

Endotoxin-inducible granulocyte-mediated hepatocytotoxicity requires adhesion and serine protease release.

In primary cultures of Kupffer cells and hepatocytes, human granulocytes potentiated toxicity of endotoxin about 1000-fold. Granulocyte elastase activity was found to correlate with toxicity. The serine protease inhibitors alpha1-antitrypsin, eglin C, and aprotinin protected against toxicity. Tumor necrosis factor-alpha (TNF-alpha) induced cytotoxicity and elastase release, whereas neutralization of TNF-alpha blocked both events. We conclude that TNF-alpha formed by Kupffer cells activates granulocytes. Experiments in cultures where cells were separated by membranes permeable to mediators indicated that cell contact is needed for toxicity. Scanning electron microscopy showed granulocytes adhering to and interdigitating with hepatocytes. Using liver cells from ICAM-1-deficient mice had no effect on toxicity. However, neutralizing CD31 inhibited toxicity and elastase release but not granulocyte adhesion. Our findings demonstrate that adhesion of granulocytes is a necessary but not sufficient condition for the synergistic interaction of endotoxin-stimulated liver macrophages and granulocytes in the proteolytic killing of hepatocytes.

The effect of prostaglandin E1 on the increase of serum lactate and plasma granulocyte elastase activity during radical surgery for esophageal cancer

Serum lactate concentrations and the lactate/pyruvate (L/P) ratio were measured in two groups of patients undergoing radical esophagectomy, as an indicator of tissue hypoxia, and β-glucuronidase and granulocyte elastase as indicators of tissue damage. One group received prostaglandin E1 (PGE1) and the other group received nothing. Serum lactate concentrations and the L/P ratio increased significantly 30 min after starting thoracotomy in the patients who were not treated with PGE1. On the contrary, intravenous drip infusion of PGE1 (0.04 μg·kg(-1)·min(-1)) suppressed the increases in serum lactate concentratons and L/P ratios. Plasma granulocyte elastase activity increased significantly at the end of surgery in both groups. There was no change in serum β-glucuronidase activity in both groups. This study suggests that low doses of PGE1 maintain organ blood flow without affecting blood pressure. However, these low doses of PGE1 could not suppress granulocyte elastase release.

Peritoneal interleukin-6, interleukin-8 and granulocyte elastase activity after elective abdominal surgery.

In this study we investigated the interleukin-8 concentration (IL-8) and granulocyte elastase activity (GE) after elective abdominal surgery. Postoperative interleukin-6 (IL-6), IL-8 concentrations and GE in the peritoneal fluid were examined in 27 patients who underwent various types of elective abdominal surgery. We compared these results with clinical parameters of surgical stress, operating time (OT) and blood loss during the operation (BL). P-IL-6 and P-IL-8 were significantly correlated with OT (P-IL-6; r = 0.67, P < 0.001: P-IL-8; r = 0.59, P < 0.001) and BL (P-IL-6; r = 0.61, P < 0.001: P-IL-8: r = 0.48, P < 0.01). P-IL-8 was significantly correlated with P-IL-6 (r = 0.68, P < 0.001) and there was a positive correlation between GE and P-IL-8 (r = 0.37, P < 0.05). These findings indicate that IL-8 might activate granulocytes in the peritoneal cavity after elective abdominal surgery and that assaying P-IL-6 and P-IL-8 is useful in assessing the host's response to surgical stress.

The effect of prostaglandin E1 on the increase of serum lactate and plasma granulocyte elastase activity during radical surgery for esophageal cancer

The effect of prostaglandin E1 on the increase of serum lactate and plasma granulocyte elastase activity during radical surgery for esophageal cancer

Altered relation between granulocyte elastase and alpha-2-macroglobulin in gingival crevicular fluid from sites with periodontal destruction.

Granulocyte elastase activity and alpha-2-macroglobulin (alpha-2-MG) were studied in the gingival crevicular fluid (GCF) from 3 categories of sites in 6 patients with gingivitis and 6 patients with periodontitis. 6 inflamed sites in each gingivitis patient were sampled on paper strips and 12 sites, 6 with and 6 without attachment loss and periodontal pockets, were selected in each periodontitis patient. To avoid the influence of increase GCF volume from deep pockets, the elastase activity and the alpha-2-MG were calculated per microliters of GCF. The proteolytic activity of elastase was measured with a low molecular weight substrate and the antiprotease, alpha-2-MG, with ELISA. The measured activity could be ascribed to elastase that had been released into the gingival tissues and into the GCF prior to sampling. In the periodontitis patients, the sites with tissue destruction had a significantly higher elastase activity per site and per microliters GCF and a significantly lower alpha-2-MG per microliters than the 2 other categories of sites without tissue destruction. The destructive inflammation seems to be associated with increased release of elastase, either from more numerous or from more active granulocytes and with an increased proteolytic consumption of the inhibitor accompanied by the fast elimination of the protease-inhibitor-complex. In conclusion, the study shows a strong relationship between elastase activity and tissue destruction, a finding that supports the pathogenic theory of an involvement of granulocytes and their proteolytic enzymes in the mechanism of periodontal destruction.

Granulocyte elastase in gingival crevicular fluid. A possible discriminator between gingivitis and periodontitis.

The granulocyte elastase activity and the immuno-reactive (antigenic) granulocyte elastase of gingival crevicular fluid (GCF) were studied in 16 periodontitis patients and in 10 gingivitis patients. The elastase activity was measured with a low molecular weight substrate specific for granulocyte elastase. The antigenic elastase was determined with specific antibodies against granulocyte elastase. Intracrevicular sampling of GCF with paper strips for 30 s seemed to provide representative values of elastase. The elastase activity correlated with probing depth and attachment loss and appeared to be a measure of the degree of tissue destruction. Antigenic elastase represents the number of granulocytes in GCF and should thus be related to the degree of inflammation. The periodontitis patients and the gingivitis patients both had a similar degree of inflammation as measured by antigenic elastase per microliter GCF and gingival index. The elastase activity per microliter GCF, however, was higher in the periodontitis group. Elevated granulocyte elastase activity in GCF seems to be independent of inflammation and could thus be an indicator of patients at risk for periodontitis.

Role of Granulocyte Elastase in Tissue Injury in Patients with Septic Shock Complicated by Multiple-organ Failure

To better understand the role of granulocyte elastase (GE) in mediating tissue injury during sepsis, GE levels were measured in plasma and bronchoalveolar lavage fluid (BALF) in patients with septic shock (n = 16) and hemorrhagic shock (n = 30). Granulocyte elastase levels were compared to levels of alpha 1-protease inhibitor (alpha 1-PI). Results show that although plasma GE-alpha 1-PI complex was initially elevated in patients with hemorrhagic and septic shock, elevations in plasma GE-alpha 1-PI complex (831 +/- 241 micrograms/L) persisted in septic shock patients. alpha 1-Protease inhibitor levels in serum were increased, resulting in an inhibition of serum GE activity. Granulocyte elastase activity in BALF, however, was significantly higher in those patients with septic, as compared to hemorrhagic shock (31.4 +/- 25.8 versus 3.7 +/- 4.0 U/L, respectively). In addition GE levels were compared to other parameters, including respiratory index, blood neutrophil count, and plasma levels of endotoxin, fibronectin, and coagulation factor XIII. Significant correlations were observed between GE-alpha 1-PI and increased endotoxin concentration and decreased fibronectin and coagulation factor XIII levels. Significant correlation was found also between GE activity in BALF and respiratory index. These findings suggest that severe tissue damage occurred in patients with septic shock complicated by multiple-organ failure. Although GE activity appeared to be adequately inhibited by alpha 1-PI in blood, increased GE activity in local tissues, such as lung alveoli, may be responsible for significant local tissue injury during septic shock.

Pregnancy-Associated Plasma Protein A Interaction with Heparin: A Critical Appraisal

Positive affinity chromatography on heparin-Sepharose has proved a most crucial step in the purification of pregnancy-associated plasma protein A (PAPP-A). In this chromatographic procedure, PAPP-A was purified almost 500-fold from term pregnancy serum. Further purification was achieved by gel filtration and negative immunoaffinity chromatography. Both PAPP-A and free heparin inhibited granulocyte elastase (HGE) activity. Whereas free heparin inhibited only in hypotonic buffers, PAPP-A inhibited HGE in hypertonic buffers also. However, PAPP-A did not inhibit other proteases (trypsin, chymotrypsin, plasmin, fibroblast collagenase) or proteolytic cascades (complement activation). Since heparin was not detected in the purified PAPP-A, the inhibition of HGE was not due to desorbed or leeched heparin ligand.

RELATIONSHIP BETWEEN RESPIRATORY DISTRESS AND GRANULOCYTE ELASTASE AND ANAPHYLATOXIN LEVELS FOLLOWING OPERATION FOR ESOPHAGEAL CARCINOMA UNDER THORACOTOMY-EFFECTS OF ULINASTATIN

This paper describes an evalation of the respiratory function and kinetics of granulocyte elastase and anaphylatoxin after operation for thoracic esophageal carcinoma, as well as a pilot study of the influence of ulinastatin, a potent protease inhibition, on the respiratory funciton. The subjects were 18 patients with thoracic esophageal carcinoma undergone radical resection, twelve of whom were allocated to ulinastatin administered group and 6 to non-administered group. Increase in granulocyte elastase activity was observed in the respiratory distress after operation for esophageal carcinoma, and it was presumed that the granulocyte elastase might be an injury causing factor to the pulmonary tissues as a result of its constituting positive coefficient correlation with respiratory index (RI). A tendency of RI being improved and inibition of granulocyte elastase were observed by administration of ulinastatin, and ulinastatin in doses sufficient to control granulocyte elastase activity inhibited a series of the reactive system which was found to be the morbidity preceding ARDS, suggesting a positive contribution of ulinastatin in the remission of respiratory distress.

RU 486 induced suppression of placental neutrophil elastase inhibitor levels

After administration of RU 486 to pregnant cynomolgus monkeys, placental morphology varied from normal to pathological. In all cases (n = 5) circulating and placental PAPP-A levels were markedly suppressed by 76.0 per cent and 55.5 per cent, respectively. When fetal demise occurred within 24 h prior to caesarian section, morphological changes consistent with an active inflammatory (polymorphonuclear leukocytosis) reaction was readily observed at the utero-placental interface, degrading the chorionic villi. Whereas heparin-Sepharose fractionated aqueous extracts of placentae inhibited human neutrophil (or granulocyte) elastase (HGE) activity, extracts of placenta being degraded by host phagocytic-proteolytic defense system were rich in HGE activity. This study establishes: (1) the cynomolgus monkey as a model for PAPP-A studies, (2) that haemochorially implanted placentae produce PAPP-A, a heparin-binding inhibitor of HGE, (3) that administration of progesterone receptor antagonist suppressed placental PAPP-A synthesis, and (4) disrupted protease-protease inhibitor equilibrium at the feto-maternal interface. Thus supporting a role for progesterone in placental PAPP-A production and maintenance of a placental barrier against maternal phagocytic-proteolytic defenses.

Effects of methylprednisolone on complement activation and leukocyte counts during cardiopulmonary bypass.

Generation of the complement activation products C3dg and terminal complement complex (TCC) and numerical changes in peripheral granulocytes (PMN) and lymphocytes were assessed in patients undergoing aortocoronary bypass surgery with extracorporeal circulation (ECC). Fluid from bronchial lavage performed preoperatively and 4 hours postoperatively was analyzed for granulocyte elastase activity and PMN content. Ten of the 20 patients received methylprednisolone (30 mg/kg b.w.) immediately before ECC. No difference was found between them and the control group regarding C3dg and TCC, and both groups showed similar postoperative decrease of peripheral blood lymphocytes. The postoperative PMN count in peripheral blood was significantly higher in the methylprednisolone group than in the controls from 12 hours onwards. In bronchial lavage fluid the postoperative PMN count was unaltered in the methylprednisolone group, but significantly increased in the controls. No granulocyte elastase activity was found before or after surgery in either group. The results indicated that methylprednisolone does not affect complement activation during cardiopulmonary bypass, but increases the granulocytes in peripheral blood postoperatively.

Stimulation of human granulocyte elastase by platelet factor 4 and heparin

Highly purified platelet factor 4 (PF 4) was found to be a potent stimulator of human granulocyte elastase activity against native elastin and solubilized α elastin. Heparin neutralized this stimulation of elastolysis by PF 4, but independently stimulated granulocyte elastase activity. Chondroitin sulfate, a constituent of the PF 4 carrier molecule, also stimulated granulocyte elastase activity. The stimulation of granulocyte elastase by PF 4 occurs at known serum concentrations of PF 4.