Hans Tegner

Quantitation Of Human Granulocyte Protease Inhibitors In Non-Purulent Bronchial Lavage Fluids

The predominant inhibitors of granulocyte proteases in plasma (alpha1-antitrypsin, alpha1-antichymotrypsin, and alpha2-macroglobulin) were quantitated in unconcentrated bronchial lavage fluids obtained from non-infected individuals, together with the acid-stable low molecular weight inhibitor with activity against granulocyte elastolytic and chymotrypsin-like enzymes. This latter inhibitor accounted for about 90% of the total molar concentration of granulocyte protease inhibitors in the bronchial lavage fluids. The remaining 10% consisted mostly of alpha1-antitrypsin and alpha1-antichymotrypsin. About 85% of the bronchial inhibitor was in a free form with preserved enzyme reactivity. The remaining 15% of the immunoreactive bronchial inhibitor exhibited a molecular size indicating complexation with enzymes. The major portion of alpha1-antitrypsin and alpha1-antichymotrypsin showed electrophoretic mobilities and molecular sizes similar to the native proteins but had no enzyme reactivity.

Glutathione in bronchoalveolar lavage fluid from smokers is related to humoral markers of inflammatory cell activity

Cigarette smoking results in variable degrees of inflammation in the lower respiratory tract. Furthermore, smoking produces oxidant-mediated changes in the lung, important to the pathogenesis of emphysema. Since glutathione can neutralize reactive oxygen species and prevent peroxidation of unsaturated lipids, it may constitute an important component of the lungs defense against oxidant and inflammatory injury. In the present study, broncholaveolar lavage (BAL) was performed in 27 smokers, and the concentrations of total glutathione as well as the cellular and humoral markers of inflammatory activity were studied. There were significant correlations between total glutathione and neutrophils; two neutrophil granule components, myeloperoxidase and elastase; and chemotactic activity for neutrophils. Moreover, the total glutathione correlated with the eosinophil cationic protein (ECP), a granule constituent of the eosinophil, with two locally produced antiproteases, secretory leukocyte protease inhibitor (SLPI) and antichymotrypsin (ACHY), but not with an α1-protease inhibitor and albumin. These data suggest that the total glutathione levels in BAL fluid may reflect a degree of oxidative and inflammatory stress caused by cigarette smoke, and they are therefore likely to contribute to the protection against this stress.

Granulocyte Collagenase, Elastase and Plasma Protease Inhibitors in Purulent Sputum

Abstract. Sputum was collected from patients with purulent chronic bronchitis. Immuno‐chemical techniques using rabbit antiserum against human granulocyte collagenase and elastase showed the presence of both enzymes. Also the serum protease inhibitors α1‐antitrypsin and α2‐macroglobulin were demonstrated. Their protease inhibiting capacity was saturated. Granulocyte elastase and collagenase occurred not only in complexes with the inhibitors, but also as free enzymes. All sputa showed free proteolytiC., elastolytic and collagenolytic activity. The concentration of collagenase was equal in the sol phase and in the gel phase of the sputa, but most of the elastase was bound to the gel phase.

Distribution of Antileukoprotease in Upper Respiratory Mucosa

Human respiratory tract secretions contain enzyme inhibitors derived from plasma and a low molecular weight, acid-stable protease inhibitor, antileukoprotease. The distribution of antileukoprotease in normal upper respiratory tract mucosa has been studied using an immunohistologic technique. The inhibitor was localized to the serous parts of the submucosal glands of the maxillary sinus and the trachea but was not demonstrable in mucous glands and goblet cells. It is concluded that the antileukoprotease found in respiratory tract secretions is produced locally in the submucosal serous glands.

Localization of Antileukoprotease in the Parotid and the Submandibular Salivary Glands

Antileukoprotease is the dominating inhibitor of granulocyte elastase and cathepsin G in normal human mixed and parotid saliva. The distribution of antileukoprotease in the submandibular and parotid glands was analysed with an immunoperoxidase technique using specific antibodies against antileukoprotease. Antileukoprotease was demonstrated in the serous cells of both the submandibular and parotid glands. These findings suggest that there is a local production of the inhibitor in the parotid gland and submandibular gland and are in agreement with our previous work which demonstrated high concentrations of the inhibitor in parotid saliva.

The effect of cigarette smoke condensate on a,‐antitrypsin, antileukoprotease and granulocyte elastase

Abstract. Unfractionated cigarette smoke condensate was found to affect α1‐antitrypsin and antileukoprotease, which are the dominating enzyme inhibitors in the respiratory tract. The electrophoretic and crossed immunoelectrophoretic precipitate patterns of these inhibitors were grossly altered and their inhibiting capacity reduced in both a dose and time dependent way. Other plasma proteins were also affected by cigarette smoke condensate, which in high concentrations caused precipitation of most serum proteins.

Interaction of Granulocyte Proteases with Inhibitors in Pulmonary Diseases

The elastase-antielastase hypothesis of lung tissue destruction has focused our interest on the two main inhibitors of granulocyte elastase in the lung, alpha 1-antitrypsin dominating blood, interstitial tissue and alveolar fluid lining and antileukoprotease dominating the respiratory tract mucosa. Antileukoprotease as well as elastase and alpha 1-antitrypsin show increased serum levels during bronchitis and bronchopneumonia, alpha 1-antitrypsin because it is an acute phase reactant, elastase and antileukoprotease because of influx from the inflamed tissues. Elastase is identified in the bronchial expectorates, mainly in complex with antileukoprotease, but often also in a free, active form. The granulocyte elastase in serum from these patients is, however, only found in complex with alpha 1-antitrypsin. The increased amounts of antileukoprotease in serum are always in a free and largely active form. The explanation for the absence of elastase-antileukoprotease complexes in serum is offered by some of our recent results. The elastase-antileukoprotease complexes are rapidly dissociated when mixed with serum in vitro, although the equilibrium dissociation constant Ki of the complex is 1.2 X 10(-11) M. Furthermore, in a pure in vitro system, alpha 1-antitrypsin is able to dissociate a leukocyte elastase-antileukoprotease complex with the rate constant of 1.3 X 10(-4) X S-1. A small part of the antileukoprotease released from the elastase-antileukoprotease complex on mixture with serum is recovered bound by elastase-alpha 2-macroglobulin complexes. Antileukoprotease inhibits the enzymatic activity of elastase-alpha 2-macroglobulin complex relatively slowly. 1:1 elastase-alpha 2-macroglobulin complexes are, however, inhibited more readily than 2:1 saturated complexes.