Elevation of Plasma Truncated Elastase α1-Proteinase Inhibitor Complexes in Patients with Inflammatory Lung Diseases

doi:10.1378/chest.102.1.129

Chest

Volume 102, Issue 1, July 1992, Pages 129-134

https://doi.org/10.1378/chest.102.1.129 Get rights and content

Human neutrophil elastase plays an important role in the development of several inflammatory lung diseases; however, there have been relatively few investigations using plasma samples. In this report, we describe alterations in the plasma elastase:α₁-PI complex in patients with chronic obstructive pulmonary disease (COPD) (15 cases), COPD with infection (8), diffuse panbronchiolitis (DPB) (8), bronchiectasis (9), pneumonia (10), and the adult respiratory distress syndrome (ARDS) (14), and in 15 normal volunteers. The elastase:α₁-PI complex concentration was determined by an enzyme-linked immunosorbent assay. Western immunoblot analysis of the elastase:α₁-PI complex was also performed. Plasma elastase:α₁-PI complex was also performed. Plasma elastase:α₁-PI complex levels in patients with COPD with infection (504μ/gL ± 93μg/L) were significantly higher, as compared with those with COPD but without infection (118μg/L ± 9μg/L) and normal volunteers (122μg/L ± 4μg/L). Increased complex concentrations were also found in patients with DPB and bronchiectasis (643μg/L ± 222μg/L and 558μg/L ± 198μg/L, respectively) as compared with normal volunteers. Increased complex concentrations were also found in patients with pneumonia and ARDS (450μg/L ± 101μg/L and 1,400μg/L ± 438μg/L, respectively). Western immunoblot analysis using anti-α₁-PI antibody and antineutrophil elastase antibody showed two types of elastase:α₁-PI complexes, one with a molecular weight of 60,000 daltons (60 kilodaltons [KD]) and the other at 50,000 daltons (50 KD). Although the native 80-KD elastase:α₁-PI complex was detected in bronchoalveolar lavage fluid, it was not found in plasma. In summary, these results demonstrated that levels of the truncated complex were increased in patients with various inflammatory lung diseases. This truncated form may play an important role in the pathophysiology of inflammatory processes. (Chest 1992; 102:129–34)

Section snippets

Patients and Controls

Plasma elastase:α₁-PI complex levels were determined in 15 normal nonsmokers, 15 patients with COPD without infection, eight patients with COPD with infection, eight with DPB, nine with bronchiectasis, ten with pneumonia, and 14 with ARDS.

The fifteen normal nonsmokers, six women and nine men with an average age of 55 yr, had no history of lung disease, and we found no clinical findings suggesting lung disease. They all had normal chest x-ray films, and their pulmonary function test results were

RESULTS

Figure 1 shows the concentrations of plasma elastase:α₁-PI complex in our study population. Data are expressed as means ± SE. Levels of plasma complexes in patients with COPD with infection (504μg/L ± 93μg/L) were significantly higher compared with COPD without infection (118μg/L ± 9μg/L; p<0.01) and normal volunteers (122μg/L ± 4μg/L; p<0.01). Increased complex concentrations were found in patients with DPB and bronchiectasis (643μg/L ± 222μg/L and 558μg/L ± 198μg/L, respectively) compared

DISCUSSION

In the present study, we measured the concentration of the plasma elastase:α₁-PI complex in patients with a variety of inflammatory lung diseases. Since epithelial inflammation is dominated by neutrophils, neutrophil elastase seemed to play an important role in the pathophysiology of lung derangements. Among the principal proteinase inhibitors in blood, α₁-PI, a 52-KD glycoprotein produced by hepatocytes and to a lesser extent by mononuclear phagocytes, is the main inhibitor, and it controls

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Association of plasma neutrophil elastase levels with other inflammatory mediators and clinical features in adult patients with moderate and severe pneumonia
2007, Respiratory Medicine
Citation Excerpt :
Plasma NE has received focus as a primary inflammatory mediator, since levels are significantly increased in patients with ARDS, sepsis, post-surgery, trauma and acute pancreatitis.5–18 However, few studies have measured plasma NE level in pneumonia,3,19,20 and the time course of plasma NE in relation to the clinical features of pneumonia has not been evaluated. Clarifying the association between the pathogenesis of pneumonia and inflammatory markers might lead to new therapeutic strategies for pneumonia and to the identification of a predictive marker for its prognosis.
Plasma levels of neutrophil elastase (NE) are elevated in several inflammatory diseases and thus this enzyme might be a critical inflammatory marker. However, the role of NE in the pathogenesis of pneumonia has not been determined. The association between the severity of pneumonia and blood levels of inflammatory markers could be relevant to developing a useful indicator of severity and new therapeutic strategies for pneumonia.
We searched for a useful predictive marker and a new therapeutic strategy against pneumonia, using a prospective, multicenter, population-based investigation. Several inflammatory markers in the circulation including NE, cytokines, defensins, C-reactive protein (CRP) and white blood cell (WBC) counts as well as clinical features were prospectively monitored in 28 adult patients with moderate (n=11) and severe pneumonia (n=17) over a period of 14 days.
The value of plasma NE was the highest at entry and significantly declined 2 days later. Trends of cytokines, defensins, CRP and WBC counts were similar but blunter. Microorganisms and the outcome of initial treatment did not significantly affect plasma NE levels. Baseline values of plasma NE were significantly higher in severe, than in moderate pneumonia and this difference between the two types of pneumonia persisted longer than those of any other markers.
Neutrophil elastase appears to play a critical role in severe pneumonia and determination of its concentration in blood could be a useful indicator of severity. Furthermore, clinical trials of anti-NE drugs in patients with severe pneumonia should be promising.
Effects of noninhibitory α-1-antitrypsin on primary human monocyte activation in vitro
2001, Archives of Biochemistry and Biophysics
A major function of α-1-antitrypsin (AAT) is the inhibition of overexpressed serine proteinases during inflammation. However, it is also known that the biological activity of AAT is affected by chemical modifications, including oxidation of the reactive-site methionine, polymerization, and cleavage by unspecific proteases, all of which will result in AAT inactivation and/or degradation. All inactive forms of AAT can be detected in tissues and fluids recovered from inflammatory sites. To test for a possible link between the inflammation-generated, noninhibitory, cleaved form of AAT and cellular processes associated with inflammation, we studied the effects of this form at varying concentrations on human monocytes in culture. We found that cleaved AAT at concentrations ranging between 1 and 10 μM in monocyte cultures over 24 h induces elevation in monocyte chemoattractant protein-1 (MCP-1) and pro-inflammatory cytokines such as TNFα and IL-6 and also increases production of interstitial collagenase (MMP-1) and gelatinase B (MMP-9), members of two different classes of matrix metalloproteinase. Moreover, monocytes stimulated with higher doses of cleaved AAT show an increase in cellular oxygen consumption by about 30%, while native AAT under the same experimental conditions inhibits oxygen consumption by about 50%. These results indicate that the cleaved form of AAT may play a role in monocyte recruitment and pro-inflammatory activation during inflammatory processes, and also suggest that changes in structure occurring upon AAT cleavage could alter its functional properties with potential pathological consequences.
Source of elastin-degrading enzymes in mycotic aortic aneurysms: Bacteria or host inflammatory response?
1999, Cardiovascular Surgery
Elastolytic matrix metalloproteinases play a central role in the development of chronic atherosclerotic aortic aneurysms, but mycotic aortic aneurysms are a distinct and unusual form of aneurysm disease caused by bacterial infection. Mycotic aortic aneurysms follow a more rapid and unpredictable course than chronic aneurysm disease and they exhibit a predilection for the suprarenal aorta, further implying unique pathophysiologic mechanisms. The purpose of this study was to examine the nature and source of elastin-degrading enzymes in mycotic aortic aneurysm. Bacterial isolates and aortic tissues were obtained from four consecutive patients undergoing surgical repair of suprarenal mycotic aortic aneurysm. Using an in vitro ³H-labeled elastin degradation assay, elastin-degrading enzyme activity was only observed in the bacteria-conditioned medium from an isolate of Pseudomonas aeruginosa. Elastin-degrading enzyme activity in the aortic tissue homogenate of this patient was abolished by the serine protease inhibitor, phenylmethylsulfonyl fluoride, but it was not suppressed by the metalloproteinase inhibitor, ethylenediamine tetraacetic acid (EDTA). In contrast, elastin-degrading enzyme activity in the bacterial-conditioned medium was decreased by about half by both phenylmethylsulfonyl fluoride and EDTA. Elastin substrate zymography revealed two phenylmethylsulfonyl fluoride-inhibitable elastin-degrading enzyme activities in the aortic tissue homogenate that corresponded to human neutrophil elastase (~30 kDa) and its stable complex with alpha 1-proteinase inhibitor (~80 kDa), but no activity attributable to Pseudomonas elastase, a 33-kDa metal-dependent enzyme. Human neutrophil elastase was readily detected throughout mycotic aortic aneurysm tissues by immunohistochemistry, but elastolytic metalloproteinases were only occasionally observed. The results of this study suggest that the elastin-degrading enzyme produced in mycotic aortic aneurysm are largely serine proteases of host neutrophil origin, rather than elastases produced by the infecting microorganisms or the macrophage-derived metalloproteinases typically observed in atherosclerotic aneurysm disease. Further studies will be needed to extend these findings to a larger number of patients with mycotic aortic aneurysm and those caused by additional microorganisms.
A leukocyte elastase inhibitor reduces thrombin-induced pulmonary oedema in the rat: Mechanisms of action
1998, Pulmonary Pharmacology and Therapeutics
The effect of a selective leukocyte elastase inhibitor, ICI 200,355, on thrombin-induced pulmonary oedema was studied in rats. Thrombin administration produced an increase in lung weight (P<0.05), wet weight/dry weight ratio (P<0.05), and relative lung water content (P<0.05). The lung weight increase was reduced by the elastase inhibitor in doses of 2000, 200 and 20 μg/kg per h (P<0.05), but not by 2 μg/kg per h. A dose of 20 μg/kg per h seems to be optimal, since 10-fold and 100-fold increases in dose did not further improve the effect. Free elastase activity in lung tissue was higher after thrombin infusion than in controls, but was not depleted by the elastase inhibitor in vivo (P<0.05). This elastase activity in the lung was, however, inhibited by the elastase inhibitor in vitro, indicating that the inhibitor can block extracellular, but not intracellular elastase activity. Thrombin infusion resulted in a significant decrease in plasma elastase inhibitory capacity (P<0.05), which was depleted by the elastase inhibitor (20 μg/kg per h) (P<0.05). Myeloperoxidase activity was significantly increased in lung tissue after thrombin infusion (P<0.05). Lung myeloperoxidase activity 5 min after thrombin infusion was not affected by the elastase inhibitor, but the inhibitor induced a further increase in myeloperoxidase as seen 90 min after thrombin infusion, indicating that the effect of this inhibitor on pulmonary oedema is not due to reduction of leukocyte infiltration in the lungs, but may partly be exerted by prevention of neutrophil destruction.
Oxidant-Antioxidant Balance in Granulocytes during ARDS: Effect of N-Acetylcysteine
1996, Chest
The production of cytotoxic oxygen radicals by activated granulocytes is a proposed mechanism of lung injury in ARDS. Protective effects of N-acetylcysteine (NAC) have been described in experimental and clinical ARDS. NAC could act in part by replenishing the intracellular stores of glutathione (GSH) in activated granulocytes, leading to detoxification of oxygen radicals produced by these cells. To test this hypothesis, 16 patients in the early phase of ARDS were randomized to receive either NAC (n=8) or placebo (n=8); granulocyte GSH, granulocyte oxygen radical production, and plasma levels of granulocyte elastase were measured in blood samples drawn sequentially within 8 h after the onset of ARDS (day 0), and then 24 (day 1), 72 (day 3), and 120 h (day 5) after the first sample; treatment with NAC or placebo was started immediately after day 0 and stopped just after day 3. Granulocyte GSH was significantly higher on days 1 and 3 when NAC was received by the patient. Unstimulated oxygen radical production, as measured ex vivo by luminol- and lucigenin-amplified chemiluminescence (CL), was higher in granulocytes from ARDS patients than from healthy control subjects, but was not influenced by NAC. The plasma levels of granulocyte elastase were five to eight times above the upper normal limit on day 0, decreased steadily until day 5, and were uninfluenced by NAC. In summary, parenteral NAC treatment started within 8 h of diagnosis increases the intracellular GSH in the granulocytes of ARDS patients without decreasing spontaneous oxidant production by these cells. The mechanisms of the protective effects of this drug previously reported in experimental and clinical ARDS remain to be established.
The value of polymorphonuclear elastase in adult respiratory distress syndrome
1995, Clinica Chimica Acta
The clinical usefulness of quantitative plasma polymorphonuclear elastase (PMN-elastase) determinations as prognostic markers of adult respiratory distress syndrome (ARDS) in polytraumatized patients was analyzed. PMN-elastase and C-reactive protein (CRP) levels were determined in 55 polytraumatized patients admitted into the Intensive Care Unit. Eight patients developed ARDS and 47 patients did not. These parameters were also analyzed in a control group (n = 34). PMN-elastase levels in ARDS cases reached significantly higher values than in patients who did not develop this syndrome (P < 0.01). We conclude that the increase in plasma PMN-elastase levels can be useful in predicting the development of ARDS in polytraumatized patients, in instituting prophylactic actions and monitoring the course of the disease in these high risk patients. This test is easily adaptable to the routine of any hospital laboratory.

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: Manuscript received April 16; revision accepted October 8.

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Chest

Elevation of Plasma Truncated Elastase α1-Proteinase Inhibitor Complexes in Patients with Inflammatory Lung Diseases

Section snippets

Patients and Controls

RESULTS

DISCUSSION

J Pediatr

J Pediatr

J Pediatr

Chest

Biochem Biophys Acta

J Biol Chem

Proteins of the cystic fibrosis respiratory tract: fragmented immunoglobulin G opsonic antibody causing defective opsopha-gocytosis

J Clin Invest

Evaluation of elastase and antielastase balance in patients with chronic bronchitis and pulmonary emphysema

Am Rev Respir Dis

Elastinolytic activity in pulmonary lavage fluid from patients with adult respiratory distress syndrome

N Engl J Med

Studies on the pathogenesis of the adult respiratory distress syndrome

J Clin Invest

Enzyme-linked immunoassay for human granulocyte elastase in complex with α1-proteinase inhibitor

“PMN-elastase assay:” enzyme immunoassay for human polymorphonuclear elastase complexed with α1-proteinase inhibitor

J Clin Chem Clin Biochem

Elevation of Plasma Truncated Elastase α₁-Proteinase Inhibitor Complexes in Patients with Inflammatory Lung Diseases

Enzyme-linked immunoassay for human granulocyte elastase in complex with α₁-proteinase inhibitor

“PMN-elastase assay:” enzyme immunoassay for human polymorphonuclear elastase complexed with α₁-proteinase inhibitor