Uptake of immunoreactive leukocyte elastase-inhibitor complexes in macrophage-like cells in abscesses.

Leukocyte elastase was demonstrated immunohistochemically not only in PMN-leukocytes in subcutaneous abscesses but also in scattered macrophage-like cells which in addition contained immunoreactive alpha 1-proteinase inhibitor (alpha 1PI) and alpha 2-macroglobulin (alpha 2M). These cells were located mainly in the marginal region of the abscess. It is concluded that the intracellular deposits in the macrophage-like cells represent phagocytized elastase and/or elastase-alpha 1PI and elastase-alpha 2M complexes. A contributing, local production of the inhibitor is, however, not excluded by the present findings.

Proteases and protease inhibitor balance in peritonitis with different causes.

Protease activation and protease-antiprotease interactions were sequentially studied in two different groups of patients with peritonitis. The biochemical changes were related to the cause of the disease and to the clinical course. Protease activation and protease inhibitor consumption were most pronounced in the peritoneal fluid, especially in bacterial peritonitis. Plasma changes also indicated activation of the complement, kinin, and fibrinolytic systems and protease inhibitor consumption, especially of alpha 2-macroglobulin and antithrombin III. There was no significant difference between chemical and bacterial peritonitis regarding these plasma changes. Immunohistologic studies showed evidence of active uptake of protease-antiprotease complexes in macrophage-like cells in the peritoneum in both groups. It is concluded that peritonitis results in protease activation and protease inhibitor consumption, especially in the peritoneal fluid. The peritoneum has an active role in the clearance of protease-antiprotease complexes. The intensity, not the type, of the intra-abdominal challenge determines the biochemical changes in peritonitis.

Abnormal proteolysis (DIC)--successful treatment with antithrombin III concentrate and a concentrate containing F XIII and native von Willebrand factor.

Two patients with life-threatening disseminated intravascular coagulation (DIC) syndrome, one caused by Gram-negative bacteria and one by premature separation of the placenta, are described. Specific substitution was given by antithrombin III concentrate and AHF-Kabi, a low purity factor VIII concentrate containing native von Willebrand factor and factor XIII. The treatment quickly returned the extremely low levels of antithrombin III, factor VIII:C, fibrinogen and factor XIII, initially found, to normal, and also returned the multimeric pattern of von Willebrand factor to normal. This resulted in diminished bleeding, enabling surgical treatment of the underlying disease.

Granulocyte collagenase and elastase and the plasma protease inhibitors in human pus.

Granulocyte collagenase and elastase was demonstrated in human pus using specific antisera. The enzymes were complexed by alpha1-antitrypsin and alpha2-macroglobulin but also were present as free proteases. All purulent exudates showed free elastolytic and collagenolytic activity.

Isolation and partial characterization of elastase from dog granulocytes.

1. An elastolytic enzyme has been isolated from dog granulocyte leukocytes. The purification procedure included preparation of the granula fraction, chromatography on Sephadex G-75 and ion-exchange chromatography on SP-Sephadex C-50 at pH 6.0. 2. The elastase isolated was homogeneous in analytical disc electrophoresis and showed in sodium dodecylsulfate electrophoresis a single protein component with the molecular weight of 24800. The enzyme lacked tyrosine and lysine and the N-terminal amino acid was phenylalanine. No carbohydrate or sialic acid were detected. 3. The dog granulocyte elastase showed similar activities as human granulocyte elastase on elastin and fibrin. The Km value for 3-carboxypropionyl-L-alanyl-L-alanyl-L-alanyl-p-nitroanilide was 2.50 mM and the pH optimum 8.5. The elastase preparation obtained was 99.5% active as judged from active-site titration. 4. The enzyme is a cationic protein and shows pronounced trailing on agarose gel electrophoresis. 5. A monospecific antiserum against the purified enzyme was produced in rabbits.

Elastases from human and canine granulocytes, I. Some proteolytic and esterolytic properties.

The lysosome-like granules of human and canine granulocytes contain an enzyme with elastinolytic activity. The enzymatic behaviour of these elastases was further characterized using the protein substrates elastin-orcein and azocasein and the synthetic substrates tert.-butyloxycarbonyl-alanine p-nitrophenylester (Boc-Ala-ONp) and 3-carboxypropionyl-L-alanyl-L-alanyl-L-alanine p-nitroanilide (Suc-Ala3-NHNp) in photometric assays. The affinities of the granulocyte elastases and of porcine pancreatic elastase to these substrates are very similar, e.g. human granulocyte elastase: KM (Boc-Ala-ONp) = 0.35mM, KM (Suc-Ala3-NHNp) = 1.25mM, porcine pancreatic elastase: KM (Boc-Ala-ONp) = 0.3mM, KM (Suc-Ala3-NHNp) - 1.15mM. The most convenient substrate for the assay of human and dog granulocyte elastases and for kinetic measurements with these enzymes is Suc-Ala3-NHNp. Using this substrate, the dissociation constant of the complex of human granulocyte elastase with human alpha1-antitrypsin could be determined (Ki = 3.5 x 10(-10)M).