alpha-chymotrypsin and Leukemia--Myeloid

Cytogenetic abnormalities are currently the most important predictors of response and clinical outcome for patients with acute myeloid leukemia (AML) or advanced-stage myelodysplastic syndrome (MDS). Because clinical outcomes vary markedly within cytogenetic subgroups, additional biological markers are needed for risk stratification.. We assessed the utility of measuring pretreatment proteasome chymotrypsin-like, caspase-like, and trypsin-like activities in plasma to predict response and survival of patients with AML (n = 174) or advanced-stage MDS (n = 52).. All three enzymatic activities were significantly (P < 0.001) increased in the plasma of patients with AML and MDS compared with normal controls. Both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, correlated with outcome. Chymotrypsin-like and caspase-like activities, but not trypsin-like activity, predicted response in univariate analysis (P = 0.002). However, only chymotrypsin-like activity was independent predictor of response from age grouping (<70 versus > or =70 years), cytogenetics, and blood urea nitrogen in multivariate analysis. Similarly, both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, were predictors of overall survival in univariate analysis (P < 0.0001), but only chymotrypsin-like activity was independent of cytogenetics, age, performance status, blood urea nitrogen, and beta(2)-microglobulin in multivariate Cox regression models. Chymotrypsin-like activity was also a strong independent predictor of survival in patients with intermediate karyotype (n = 124).. Measuring plasma chymotrypsin-like activity may provide a powerful biomarker for risk stratification in patients with AML and advanced-stage MDS, including those with normal karyotype.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Caspases; Chymotrypsin; Female; Humans; Leukemia, Myeloid; Male; Middle Aged; Multivariate Analysis; Myelodysplastic Syndromes; Neoplasm Staging; Prognosis; Proteasome Endopeptidase Complex; Risk Factors; Survival Analysis; Trypsin; Young Adult

A case of chronic monocytic leukemia with specific dermal lesions is presented. The patient developed an adenocarcinoma of the cecum.

Topics: Aged; alpha 1-Antichymotrypsin; Chymotrypsin; Female; Humans; Immunoenzyme Techniques; Leukemia, Myeloid; Microscopy, Electron; Monocytes; Skin; Skin Neoplasms

Blood and bone marrow samples from 20 individuals with reactive conditions and 26 cases of acute and chronic myeloid leukemias were tested for the presence of lysozyme, alpha-1-antitrypsin (alpha-1-AT), and alpha-1-antichymotrypsin (alpha-1-ACT). We compared the reactivity of samples in smears, cytocentrifuge preparations, and paraffin sections. Lysozyme, alpha-1-antitrypsin, and alpha-1-antichymotrypsin were found only in polymorphonuclear leukocytes and monocytes and their precursors. Lymphocytes, E-rosetting cells, Con A-activated lymphocytes, natural killer (NK) cells, red blood cells, erythroblasts, and megakaryocytes were consistently negative. Leukemic myeloblasts showed definite reactivity for both alpha-1-antitrypsin and alpha-1-ACT, but not for lysozyme. By contrast, lysozyme was present in poorly differentiated leukemic monoblasts, while alpha-1-antitrypsin and alpha-1-antichymotrypsin showed only weak reactivity. More mature myeloid and moncytic cells showed positive staining for all three antigens tested with differences in staining distribution and intensity. In four cases of chronic myeloid leukemia (CML), circulating mature polymorphonuclear leukocytes were deficient in both lysozymne and alpha-1-antitrypsin. The use of lysozyme, alpha-1-antitrypsin, and alpha-1-antichymotrypsin identifies normal and leukemic cells of the myeloid-monocytic series at all stages of maturation and is applicable to a variety of sample preparations.

Topics: alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Bone Marrow; Cell Separation; Chymotrypsin; Clinical Enzyme Tests; Flow Cytometry; Hematopoietic Stem Cells; Humans; Immunoenzyme Techniques; Killer Cells, Natural; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Microtomy; Monocytes; Muramidase; Paraffin

Two cytochemical methods for detection of granulocytic elastase and chymotrypsin employing alanine and phenylalanine naphthyl esters were developed. Specificity of reaction with the ester substrates was proven by chloromethyl ketone inhibitors. The results of both staining methods were almost identical with the staining for naphthol AS-D chloroacetate (Cl Ac-O Nap AS-D) esterase, since Cl Ac-O Nap AS-D also reacts with granulocyte elastase and chymotrypsin. Mature neutrophils and myeloid precursors except myeloblasts are stained with all three substrates in peripheral blood and bone marrow. Mast cells, however, only react with Cl Ac-O Nap AS-D and the chymotrypsin substrate and not with the elastase substrate. In acute myeloid leukemia the three esterases appear in parallel at a somewhat later stage of maturation than myeloperoxidase. In blood smears from 380 hospital patients no hereditary elastase or chymotrypsin deficiency could be demonstrated. Staining for elastase and chymotrypsin was also normal in hereditary myeloperoxidase deficiency and chronic granulomatous disease. On the other hand 6% of the hospital patients and about two-thirds of patients with acute myeloid leukemia showed a partial elastase deficiency in more than 25% of the peripheral neutrophils.

Topics: Chymotrypsin; Granulocytes; Granulomatous Disease, Chronic; Humans; Hydrogen-Ion Concentration; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Naphthol AS D Esterase; Pancreatic Elastase; Peroxidase; Phenylalanine

To show whether direct proteolysis of coagulation factors may play a role in patients with so-called consumption coagulopathy, granulocytic neutral proteases in the plasma of patients with acute myelocytic leukemia and septicemia were assayed by one- and two-dimensional Laurell electrophoresis. Complexes between serum alpha1-antitrypsin and elastase-like granulocytic protease could be demonstrated in those patients with acute myelocytic leukemia and septicemia who also had moderate or severe coagulation defects. Despite the presence of a high antiprotease potential, addition of the elastase-like enzyme to normal plasma resulted in coagulation defects in vitro comparable to those seen in the patients. These results and the ability of the elastase-like protease to destroy isolated clotting factors suggested that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to disseminated intravascular coagulation may be operational.

Topics: alpha 1-Antitrypsin; Blood Coagulation Disorders; Chymotrypsin; Electrophoresis, Agar Gel; Factor XIII; Fibrin Fibrinogen Degradation Products; Granulocytes; Humans; Immunoelectrophoresis, Two-Dimensional; Leukemia, Myeloid; Leukocytes; Pancreatic Elastase; Peptide Hydrolases; Sepsis

Three cationic proteins from the granules of human neutrophil granulocytes were obtained in a high degree of purity be means of affinity chromatography on 4-phenylbutylamine-Sepharose. Together with lysozyme, the three cationic proteins exhibit the highest electrophoretic mobility toward the cathode in acrylamide gels at moderately acid pH, among the granule constituents that are solubilized in 0.1 M phosphate buffer, pH 7.0, containing 1 M NaCl. The three cationic proteins represent a group of "neutral proteases" distinct from elastase and collagenase. They hydrolyze casein, azocasein and the chymotrypsin substrate N-acetyl-L-tyrosine ethyl ester. Optimal activity is found at pH 7.4-7;5. The enzymes are inhibited by the specific chymotrypsin inhibitor N-tosyl-L-phenylalanylchloromethane and by the naturally occurring inhibitors alpha-antichymotrypsin, alpha-1-antitrypsin, as well as by the trypsin inhibitors from soy beans and limabeans.

Topics: Blood Proteins; Chromatography, Affinity; Chymotrypsin; Electrophoresis, Polyacrylamide Gel; Granulocytes; Humans; Hydrogen-Ion Concentration; Kinetics; Leukemia, Myeloid; Leukocytes; Neutrophils; Tosylphenylalanyl Chloromethyl Ketone

Previous studies have demonstrated that some chronic myelogenous leukemia cells contain a macromolecular binding factor for folic acid. This binder, which previously was believed to be a single factor, has now been resolved into two distinct binding proteins. Separation of each binder was obtained by DEAE chromatography of the partially purified lysate of chronic myelogenous leukemia cells. One binder has a molecular weight of 30;000-35,000, and the second binder has a molecular weight of 40,000-45,000. Both proteins bind the mono-, di-, and triglutamates of folic acid, N10-methyl-folate, dihydro-folate, and N5-methyltetrahydrofolate. Neither binder has determinants for N5-formyltetrahydrofolate or methotrexate. The preferred substrates for both binders appear to be the fully oxidized and partially reduced folates rather than the fully reduced folates. The lower-molecular-weight folate binding protein shows reversible binding with partially and fully reduced folates but irreversible binding with oxidized folates. This property suggests that this binder may have some function in the transport and storage of folate. The higher-molecular-weight folate binding protein, however, has only slight reversibility of binding with the partially and fully reduced folates, and it is therefore more difficult to postulate a physiologic function for this binding factor.

Topics: Carrier Proteins; Cell-Free System; Chromatography, DEAE-Cellulose; Chymotrypsin; Deoxyribonucleases; Folic Acid; Humans; Leukemia, Myeloid; Leukocytes; Molecular Weight; Pepsin A; Ribonucleases; Trypsin

Topics: Biological Availability; Cell Division; Cells, Cultured; Chromatography, Thin Layer; Chymotrypsin; Clone Cells; Culture Media; Hematopoietic Stem Cells; Humans; Iodine Radioisotopes; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Molecular Weight; Trypsin

Topics: Aprotinin; Chromatography, Affinity; Chymotrypsin; Electrophoresis, Polyacrylamide Gel; Humans; Leukemia, Myeloid; Macroglobulins; Neutrophils; Peptide Hydrolases; Trypsin Inhibitors

Topics: Acetates; Anhydrides; Chymotrypsin; Cytoplasmic Granules; Electrophoresis, Polyacrylamide Gel; Esterases; Esters; Humans; Hydrogen-Ion Concentration; Hydrolysis; Leukemia, Myeloid; Leukocytes; Naphthols; Pancreatic Elastase; Trypsin; Tyrosine

Topics: Antigens; Chymotrypsin; Enzyme Precursors; Humans; Leukemia; Leukemia, Myeloid; Spleen