Serum Neutrophil Elastase Research Articles

Neutrophil elastase inhibitor improves postoperative clinical courses after thoracic esophagectomy

Sivelestat sodium hydrate is a selective inhibitor of neutrophil elastase (NE), and is effective in acute lung injury associated with systemic inflammatory response syndrome (SIRS). The effect of Sivelestat for postoperative clinical courses after transthoracic esophagectomy was investigated. Consecutive patients with carcinoma of the thoracic esophagus who underwent transthoracic esophagectomy between 2003 and 2004 were assigned to the Sivelestat-treated group (n = 18), and those between 1998 and 2003 were assigned to the control group (n = 25). The morbidity rate, duration of postoperative SIRS, mechanical ventilation, and intensive care unit (ICU) stay, and the sum of the sequential organ failure assessment scores at all time points after the operation were compared. Serum NE activities and serum concentrations of TNF-alpha, IL-1beta, IL-6, and high mobility group box chromosomal protein 1 (HMGB1) were measured. Postoperative complications developed in three patients in the control group, and one in the Sivelestat-treated group. The durations of SIRS, mechanical ventilation, and ICU stay were significantly shorter in the Sivelestat-treated group. Even in patients without complications, the durations of mechanical ventilation, and ICU stay were also significantly shorter, and the arterial oxygen pressure/fraction of inspired oxygen ratio at postoperative day 1 was significantly higher in the Sivelestat-treated group. Serum NE activities and serum concentrations of IL-1beta, IL-6, and HMGB1 were significantly suppressed in the Sivelestat-treated group. Postoperative Sivelestat treatment after transthoracic esophagectomy improves the condition of SIRS and postoperative clinical courses, even in patients without complications.

Overexpressed circulating serine proteases are a predominant source of self-antigen for potent T lymphocyte immunity to human leukemia (48.5)

Abstract From the experience of clinical transplantation, cytotoxic T lymphocytes (CTL) mediate strong anti-leukemia immunity, although a little is known about the targets of that immunity. We identified an HLA-A2- restricted nonomer PR1 (VLQELNVTV) from proteinase 3 (P3) and neutrophil elastase (NE) that was recognized by CTL, that preferentially killed leukemia cells. To determine the protein source of PR1, we studied distribution of both P3 and NE in patients with AML and CML. Serum NE (ng/ml) assessed by ELISA was increased in 13 out of 14 AML patients and in 19 out of 20 CML patients, compared to that in 4 healthy donors (HD) (1055 ± 132, and 1002 ± 103 versus 54 ± 13; p = 0.0002 and p = 0.0006, respectively). In contrast, overexpression of serum P3 (ng/ml) in 19 AML patients and 28 CML patients was not significantly different compared to the mean from 14 HD (33 ±18 and 56 ± 21 versus 10 ± 2.2; p = 0.13 and p = 0.08, respectively). In nine myeloid leukemia patients the marrow-derived P3 level, which was similar to serum P3, was also similar to that in five non-myeloid leukemia patients (23 ± 9.5 versus 27 ± 11.8, p = 0.08). Further P3 was highly expressed on plasma membrane (PM) of both marrow-derived and circulating blasts, whereas NE was only expressed on PM of marrow-derived blasts. Therefore aberrant expression of serine proteases in leukemia may play a role in reversing T cell tolerance by increasing available determinants on MHC-I in leukemia.

Elevated Serum Neutrophil Elastase but Not Proteinase 3 Is Present in Patients with Myeloid Leukemia.

Proteinase 3 (P3) and neutrophil elastase (NE) are abnormally expressed in the cytoplasm of myeloid leukemia, and we have shown that the HLA-A2-restricted peptide PR1 is processed and cross-presented from both proteins by antigen-presenting cells (APC) to cytotoxic T lymphocytes (CTL). PR1-specific CTL kills leukemia cells and contributes to cytogenetic remission in patients with chronic myelogenous leukemia. Both exogenous P3 and NE can bind to monocytes and P3 promotes DC maturation in vitro. Since high levels of circulating P3 were found in patients with the ANCA-associated systemic vasculitis Wegener's granulomatosis, modulation of DC by P3 has been implicated in reversal of tolerance. The role of extracellular P3 and NE in reversing tolerance in patients with myeloid leukemia is unknown. We used an ELISA assay to assess P3 and NE level in the sera of patients with AML and CML. Serum NE level was highly increased in all 15 AML patients studied (1045±149 ng/ml, n = 15) and in all 15 CML patients (1013±128, n = 15), compared to a control group of healthy donors (54±13 ng/ml, n = 4; p = 0.0038 and p = 0.0014, respectively). In contrast, serum P3 was increased in only 5 out of 18 (28%) AML patients (47 ± 18, n = 18 ng/ml) and in 8 out of 28 (29%) CML patients (95 ± 44 ng/ml, n = 28) compared to a group of 14 healthy donors (10 ± 2.2 ng/ml; p = 0.09 and p = 0.06, respectively). There was no relationship between elevated serum NE and P3 levels (r=0.16, n=35). P3 level in untreated patients and healthy donors did not change within at least 13 months, although P3 decreased after allogeneic BMT in 5 out of 5 patients (p < 0.05) with high serum levels of P3 pre-BMT. Because P3 is expressed by myeloid progenitor cells in the bone marrow, we studied the concentration of P3 in bone marrow-derived serum (BMS). To our surprise, BMS P3 concentration in patients with myeloid leukemia (23 ± 9.5 ng/ml, n = 9) was similar to that of a control group of patients with non-myeloid hematological malignancies (27 ± 11.8, n = 5) (p = 0.08), and BMS P3 concentration was similar to sera P3 concentration in the same patient (p = 0.52). Interestingly, BMS P3 level correlated with the level of BMS GM-CSF (quantified by ELISA) in a small group of patients with AML and CML (r = 0.94, n = 9), suggesting that P3 secretion by leukemia cells may be downstream of GM-CSF. We also analyzed membrane expression of P3 and NE in 5 AML patients using flow cytometry and found that P3, but not NE, was expressed on circulating cells in 3 of 5 patients. This unique expression of P3 and NE suggests that circulating NE might be the predominant source of cross-presented PR1 antigen by APC or myeloid leukemia cells.

アルコール依存症と摂食障害による低栄養状態に重症肺炎による急性呼吸窮迫症候群を合併したがシベレスタットナトリウムの早期導入により血清好中球エラスターゼ活性値の改善を認め救命しえた若年女性の1例

A 30-year-old woman with malnutrition due to alcoholism and eating disorders was found to have acute respiratory distress syndrome (ARDS) and sepsis due to severe Streptococcus pneumoniae pneumonia. S. pneumoniae was detected by an in vitro rapid immunochromatographic assay for S. pneumoniae antigen in urine on the day of admission and by blood culture 2 days after admission. Symptoms and laboratory findings improved after treatment with sivelestat sodium hydrate, antibiotics, and mechanical ventilation. Treatment with sivelestat sodium hydrate also decreased serum neutrophil elastase activity. This case demonstrates the usefulness of early treatment with sivelestat sodium hydrate in ARDS due to severe pneumonia.

Neutrophil‐mediated inflammation in respiratory syncytial viral bronchiolitis

The involvement of neutrophil-mediated inflammation may play an important role in the pathogenesis of acute respiratory syncytial virus bronchiolitis. However, no measurable marker is sensitive enough to assess neutrophil-mediated inflammation in the airways. Released neutrophil elastase (NE) in intraluminal airways has been reported to induce pulmonary inflammation. The aim of this study was to determine whether the amount of urinary trypsin inhibitor (UTI) in serum, a degenerate induced by NE, reflects the degree of airway inflammation in children with respiratory syncytial viral (RSV) bronchiolitis and whether the severity of inflammation is evaluated. The pre-alpha-/inter-alpha-trypsin inhibitor is assumed to be precursors of the UTI. When NE degrades these inhibitors, UTI is liberated. Serum UTI concentrations in infants admitted with RSV bronchiolitis, other viral infections, bacterial pneumonia and control subjects were measured by means of one-step sandwich-type enzyme immunoassay. Serum UTI concentrations in 25 patients on admission were significantly higher than the 15 infantile control values (mean +/- SEM, 22.126 +/- 2.317 and 6.701 +/- 0.719 U/mL, respectively; P < 0.0001). The elevated levels returned to baseline values with improvement in the respiratory symptoms. Higher levels of serum UTI with RSV infection were consistently associated with clinical symptoms and artificial ventilation. Serum NE concentrations of patients were elevated in some patients but not significantly different from controls in the patients who showed only upper respiratory symptoms with RSV infections. The findings strongly suggested that neutrophil-mediated events are involved in the pathogenesis of RSV bronchiolitis, and the monitoring of UTI concentrations might be useful for evaluating the neutrophil-mediated airway inflammation.

Increased serum concentration of urinary trypsin inhibitor with asthma exacerbation.

The aim of the study was to determine whether the amount of urinary trypsin inhibitor (UTI) in serum, a degenerate induced by neutrophil elastase (NE), reflects the degree of bronchial inflammation in children with acute asthma exacerbation. The involvement of neutrophil-mediated inflammation plays as important a role as eosinophil-mediated inflammation in the pathogenesis of acute asthma exacerbation. However, no measurable marker is sensitive enough to assess neutrophil-mediated inflammation in the airways. The pre-alpha-/inter-alpha-trypsin inhibitors are assumed to be precursors of UTI. NE degrades pre-alpha-/inter-alpha-trypsin inhibitors to liberate UTI. UTI concentrations in 25 childhood patients admitted with asthma exacerbation and 15 control subjects were measured by means of one-step sandwich-type enzyme immunoassay. Serum UTI concentrations in the patients at admission were significantly higher than control values (10.597+/-0.649 and 6.136+/-0.303 U x mL(-1), respectively (mean+/-SEM)). These levels returned to baseline values with improvement in the asthmatic symptoms. However, serum NE and alpha1 antitrypsin concentrations were not significantly different between patients and controls, even during acute exacerbation in the former. The findings suggest that neutrophil-mediated inflammatory events are involved in exacerbation of childhood asthma. The monitoring of urinary trypsin inhibitor concentrations might be useful for evaluating the neutrophil-mediated inflammation in childhood asthma attack.

Pulmonary endothelial permeability changes afte major lung resection

Increased pulmonary endothelial permeability has been proposed as a cause of postpneumonectomy pulmonary edema. This study investigated changes in pulmonary endothelial permeability after major lung resection. Lung scintigraphy was performed in 21 men (median age, 66 years; range, 34 to 73 years) after pneumonectomy (10 patients) or lobectomy (11 patients). Pulmonary endothelial permeability was measured by the net pulmonary accumulation of intravenous technetium-99m-labeled albumin, calculated as a ratio of lung:heart radioactivity counts. Pulmonary hemodynamics were monitored continuously by a pulmonary artery catheter, and serum levels of inflammatory cytokines were assayed. The lung:heart radioactivity ratio increased significantly in the initial 8 hours after pneumonectomy but not after lobectomy (p < 0.01). Mean pulmonary artery pressure and pulmonary vascular resistance both increased significantly during pneumonectomy (p < 0.05). The intraoperative increase in mean pulmonary artery pressure was inversely related to preoperative mean pulmonary artery pressure (r = -0.47; p = 0.02). The postoperative change in lung:heart radioactivity ratio to the perioperative increase in pulmonary vascular resistance (r = 0.54; p = 0.02) but not to the increase in mean pulmonary artery pressure (r = 0.14; p > 0.05). Serum interleukin-8 and neutrophil elastase levels were elevated in all patients preoperatively. The postoperative change in lung:heart radioactivity ratio was related to preoperative elastase levels (r = 0.61; p = 0.02). Pulmonary endothelial permeability appears to be increased after pneumonectomy. Preoperative neutrophil activation and the adaptation of the remaining pulmonary vasculature may be etiologic factors.

Vasculitis and antineutrophil cytoplasmic autoantibodies associated with propylthiouracil therapy

Vasculitis is a rare complication of propylthiouracil therapy. Antineutrophil cytoplasmic antibodies (ANCA) have been described in association with several vasculitic disorders. We report detection of ANCA against human neutrophil elastase, proteinase 3, and myeloperoxidase in serum from six patients who developed evidence of vasculitis during propylthiouracil treatment of hyperthyroidism. On withdrawal of the drug ANCA concentrations fell and clinical symptoms resolved completely.

Serum Antielastase and Neutrophil Elastase Levels in PiM Phenotype Cigarette Smokers with Airflow Obstruction

In order to assess blood factors which might explain why some cigarette smokers develop airflow obstruction while others do not, we compared two groups of PiM phenotype volunteers matched for age, sex and total pack-years of cigarette smoking; one group had airflow obstruction and the other did not. Functional levels of alpha-2-macroglobulin (alpha-2-M) and alpha-1-protease inhibitor (alpha-1-PI) were separately assessed by a protease binding procedure. Neutrophils were isolated from blood by counterflow centrifugation, and their elastase content was assayed with 3H-elastin-SDS (sodium dodecyl sulfate). The obstructed and nonobstructed groups were not different with respect to functional or immunoreactive levels of alpha-1-PI and alpha-2-M or elastase levels in their neutrophils. We do not find imbalances of circulating elastase or antielastase levels in PiM phenotype smokers with airflow obstruction.