lactoferrin and Leukemia--Myeloid--Acute

Topics: Cell Differentiation; Cell Division; Cytokines; Gene Expression Regulation, Leukemic; Humans; Lactoferrin; Leukemia, Myeloid, Acute; Muramidase; Myeloblastin; Peroxidase; Serine Endopeptidases

Topics: Adolescent; Adult; Aging; Bone Marrow Transplantation; Child; Child, Preschool; Humans; Immunoenzyme Techniques; Indicators and Reagents; Infant; Infant, Newborn; Lactoferrin; Leukemia, Myeloid, Acute; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Reference Values

Whereas the diagnosis of acute lymphoid leukaemia greatly depends on immunophenotyping on the leukaemic cells, the diagnosis of acute myeloid leukaemia (AML) is still only based on morphological and cytochemical criteria. Here we describe that with a monoclonal antibody, directed against myeloperoxidase (MPO), the immunological diagnosis of AML is possible in most cases. A monoclonal antibody against lactoferrin (LF) was used to detect more mature myeloperoxidase-containing cells. Of the cell samples tested from 206 different patients with AML, 95% were found to express myeloperoxidase in more than 15% of lactoferrin-negative cells. Compared with other myeloid-reactive monoclonal antibodies (VIM2, anti-CD13, anti-CD14, anti-CD15 and anti-CD33), a higher diagnostic sensitivity and specificity for AML was found. No significant correlation with the FAB classification was found. In most patients, more MPO-positive cells were detected by the monoclonal antibody than by the cytochemical staining. This could be due to the recognition of enzymatically inactive precursor forms of myeloperoxidase by the antibody. The use of anti-myeloperoxidase monoclonal antibodies for the diagnosis of AML has the advantage that objective quantification is possible.

Topics: Antibodies, Monoclonal; Bone Marrow; Humans; Lactoferrin; Leukemia, Myeloid, Acute; Peroxidase

The prevention of enterogenic infection by human lactoferrin was tested in five neutropenic patients receiving chemotherapy for acute myelogenous leukemia. Lactoferrin did not significantly delay the onset of infection but reduced its duration and severity as judged from the course of fever. Compared with nine matched controls, lactoferrin-treated patients had a lower incidence of bacteremia on the whole and of gram-negative bacteremia in particular.

Topics: Adolescent; Adult; Agranulocytosis; Female; Fever; Humans; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Male; Middle Aged; Neutropenia; Sepsis

Topics: Gene Expression Regulation, Neoplastic; Humans; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Neoplastic Stem Cells; RNA, Messenger

Neutrophils and band forms from patients with acute myeloid leukemia and myelodysplastic syndrome were stained for the presence of myeloperoxidase using a cytochemical method (diaminobenzidine/hydrogen peroxide) and the alkaline phosphatase--anti-alkaline phosphatase immunocytochemical procedure (using monoclonal anti-myeloperoxidase). Neutrophils and bands were also stained for elastase and lactoferrin using monoclonal and polyclonal antibodies, respectively. Subpopulations of neutrophils and bands from cases of acute myeloid leukemia and myelodysplasia exhibited a qualitative and/or quantitative deficiency in myeloperoxidase. In addition, a quantitative decrease in elastase and/or lactoferrin staining was detected. Thus, neutrophils and bands from patients with acute myeloid leukemia and myelodysplastic syndrome have a defect in one or more of the constituents of primary and/or secondary granules. These defects are consistent with the view that abnormal neutrophils and bands are derived from a malignant clone of myeloid precursor cells.

Topics: Cytoplasmic Granules; Humans; Immunohistochemistry; Lactoferrin; Leukemia, Myeloid, Acute; Myelodysplastic Syndromes; Neutrophils; Pancreatic Elastase; Peroxidase

A panel of eight antibodies was used by the alkaline-phosphatase/anti-alkaline phosphatase (APAAP) method to stain peripheral blood films, bone marrow smears, and cytocentrifuge preparations from 29 cases of acute myeloid leukemia. These findings were correlated with the French-American-British (FAB) classification. Leukemic cells from six cases of myeloblastic leukemia (FAB;M1) were predominantly labeled by the antimyeloperoxidase monoclonal antibody (MPO-7). Leukemic cells from the majority of eight cases of myeloblastic leukemia with maturation (FAB;M2) and progranulocytic leukemia (FAB;M3) stained with monoclonal antibodies MPO-7, NP57 (anti-elastase), and EBM11 (antimonocyte/macrophage). Leukemic cells from six cases of myelomonocytic (FAB;M4) and five cases of monocytic (FAB;M5) leukemia were most often labeled with antibodies MPO-7, NP57, and EBM11 as well as monoclonal antibodies Y1/82A (anti-monocyte) and KB90 (against the p150, 95 molecule, CD11c; a monocyte/granulocyte marker), but not with monoclonal antibody C17 (antiglycoprotein IIb/IIIa) and/or monoclonal antibody Y2/51 (antiglycoprotein IIIa). Erythroblasts from a single case of erythroleukemia (FAB;M6) were not labeled with any of the antibodies from this panel. Leukemic cells from two cases of acute megakaryocytic leukemia (FAB;M7) stained strongly with the monoclonal antiglycoprotein IIIa/IIb antibody (C17) and antiglycoprotein IIIa antibody (Y2/51). Staining by the APAAP method with this panel of antibodies was easy to perform, required no expensive instrumentation, and provided useful information in the classification of acute myeloid leukemia.

Topics: Alkaline Phosphatase; Antibodies, Monoclonal; Histocytochemistry; Humans; Lactoferrin; Leukemia, Myeloid, Acute; Macrophages; Monocytes; Peroxidase; Phenotype; Platelet Membrane Glycoproteins

92 patients with acute myeloid leukemia were classified according to the FAB classification (M1 n = 20, M2 n = 43, M3 n = 1, M4 n = 19, M5a n = 2, M5b n = 2, and M6 n = 5 patients). Serum measurements of lactoferrin (LF), myeloperoxidase (MPO) and lysozyme (LYS) were performed before the start of treatment. LF was significantly lower in M1 when compared with M2 but not as compared to M4, MPO was significantly higher in M2 and M4 than in M1, but comparable MPO levels were found in M2 and M4. LYS was significantly elevated in M2 in comparison with M1, and in M4 when compared to both M1 and M2. Polymorphonuclear granulocytes (PMNs) in M1 were significantly reduced when compared with M2 and M4, whereas mononuclear cells were significantly increased in M4 in comparison with both M1 and M2. FAB classification did not generate any prognostic information. When the patients were, instead, subdivided according to LF levels were found prognostically significant differences. Of patients below 100 micrograms/l, 44% went into remission as compared to 77% with LF from 101 to 400 micrograms/l. In patients with LF levels above 400 micrograms/l the remission frequency was only 14%. Multivariate statistical analysis on the data further suggested that lactoferrin may be used as an independent prognostic indicator. We conclude that although determination of the serum-levels of lactoferrin, lysozyme and myeloperoxidase in certain cases may be valuable as a supplement to the morphological examination of acute myeloid leukemia, it is evident that none of the three determinations can be used alone to distinguish between the FAB groups.

Topics: Adult; Aged; Female; Genetic Variation; Granulocytes; Humans; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Male; Middle Aged; Muramidase; Peroxidase; Prognosis

Serum levels of lactoferrin, lysozyme and myeloperoxidase were measured sequentially after induction treatment in 30 patients with AML in order to test the hypothesis that these proteins may be used to monitor activity and different stages of myelopoiesis in the bone-marrow. The results showed that myeloperoxidase and lysozyme started to rise again 6-8 days after initiation of treatment as compared to 12 d for lactoferrin and 16 d for blood polymorphonuclear leukocytes. The rate of marrow regeneration was exponential and estimated to be 9-20% per d. The comparison between two different chemotherapy regimes showed that the initial reduction in lysozyme, in contrast to other measured variables, was significantly larger with the therapy that contained vincristine and glucocorticosteroids. This might reflect a reduction in lysozyme secretion in addition to the effects on the leukemic cell mass. We conclude that the measurements of lactoferrin, lysozyme and myeloperoxidase in serum under certain circumstances may be used to monitor the myelopoietic activity in the bone-marrow.

Topics: Antineoplastic Combined Chemotherapy Protocols; Bone Marrow; Cytarabine; Daunorubicin; Doxorubicin; Hematopoiesis; Humans; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Leukocyte Count; Muramidase; Neutrophils; Peroxidase; Prednisolone; Thioguanine; Time Factors; Vincristine

This study describes an ELISA technique with high specificity, sensitivity, accuracy and reproducibility for measurements of plasma lactoferrin. The detection limit was 0.001 microgram/ml and the median value obtained in EDTA plasma from 47 healthy adults was 0.100 microgram/ml (0.05 fractile: 0.046 microgram/ml, 0.95 fractile: 0.257 microgram/ml). The lactoferrin concentration in serum was on the average 2 1/2 times higher than in plasma. The ambient temperature did not influence the plasma concentration during the first 6 h from blood sampling to separation of plasma from the cells. In 8 patients with untreated acute leukaemia plasma lactoferrin was positively correlated to the peripheral neutrophil count. An almost parallel course in plasma lactoferrin and peripheral neutrophil number was observed in 4 patients with AML during chemotherapy. In 2 patients achieving complete remission, plasma lactoferrin increased about 6 d before the concomitant increase in neutrophil count, suggesting plasma lactoferrin as an early predictor of bone marrow regeneration.

Topics: Adolescent; Adult; Aged; Bone Marrow; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immune Sera; Kinetics; Lactoferrin; Lactoglobulins; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Leukocyte Count; Male; Middle Aged; Neutrophils; Temperature

Delivery of iron to K562 cells by diferric transferrin involves a cycle of binding to surface receptors, internalization into an acidic compartment, transfer of iron to ferritin, and release of apotransferrin from the cell. To evaluate potential feedback effects of iron on this system, we exposed cells to iron chelators and monitored the activity of the transferrin receptor. In the present study, we found that chelation of extracellular iron by the hydrophilic chelators desferrioxamine B, diethylenetriaminepentaacetic acid, or apolactoferrin enhanced the release from the cells of previously internalized 125I-transferrin. Presaturation of these compounds with iron blocked this effect. These chelators did not affect the uptake of iron from transferrin. In contrast, the hydrophobic chelator 2,2-bipyridine, which partitions into cell membranes, completely blocked iron uptake by chelating the iron during its transfer across the membrane. The 2,2-bipyridine did not, however, enhance the release of 125I-transferrin from the cells, indicating that extracellular iron chelation is the key to this effect. Desferrioxamine, unlike the other hydrophilic chelators, can enter the cell and chelate an intracellular pool of iron. This produced a parallel increase in surface and intracellular transferrin receptors, reaching 2-fold at 24 h and 3-fold at 48 h. This increase in receptor number required ongoing protein synthesis and could be blocked by cycloheximide. Diethylenetriaminepentaacetic acid or desferrioxamine presaturated with iron did not induce new transferrin receptors. The new receptors were functionally active and produced an increase in 59Fe uptake from 59Fe-transferrin. We conclude that the transferrin receptor in the K562 cell is regulated in part by chelatable iron: chelation of extracellular iron enhances the release of apotransferrin from the cell, while chelation of an intracellular iron pool results in the biosynthesis of new receptors.

Topics: Apoproteins; Cell Line; Cell Membrane; Deferoxamine; Humans; Iron; Iron Chelating Agents; Kinetics; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Pentetic Acid; Receptors, Cell Surface; Receptors, Transferrin; Transferrin

Blood neutrophils were studied by quantitative cytochemistry in patients with acute myeloid leukemia at diagnosis (17 patients), during remission (17 patients) and in relapse (seven patients). Scanning and integrating microdensitometry was used to quantify components of azurophilic granules (myeloperoxidase, chloroacetate esterase, beta-glucuronidase, acid phosphatase, acid mucosubstance) and also specific granules (lactoferrin). At diagnosis, neutrophil myeloperoxidase, chloroacetate esterase, and lactoferrin were significantly decreased, compared with normal neutrophils from 25 controls, with 13 of the 17 patients showing a partial or complete deficiency of at least one granule constituent. Five of seven patients, followed serially from remission into relapse, showed a fall in activity of azurophilic or specific granule components before overt blast cell infiltration of the marrow had occurred and this may predict relapse.

Topics: Acid Phosphatase; Adult; Carboxylic Ester Hydrolases; Cytoplasmic Granules; Glucuronidase; Histocytochemistry; Humans; Lactoferrin; Leukemia, Myeloid, Acute; Neutrophils; Peroxidase; Time Factors

Topics: Child; Granulocytes; Hematopoietic Stem Cells; Humans; Lactoferrin; Lactoglobulins; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Peroxidase; Peroxidases; Reference Values

Semiquantitative analysis of lactoferrin deficiency in neutrophil polymorphonuclear leucocytes in various haematological and non-haematological disease was carried out by scoring polymorphonuclear leucocytes stained for lactoferrin by the immunoperoxidase method. The staining patterns for lactoferrin were classified into four types (0-III) based on the intensity of reaction, and the sum of the ratings of 100 polymorphonuclear leucocytes was considered as "lactoferrin score" with a possible range of 0-300. As a result, significantly low lactoferrin-scores were frequently observed in acute leukaemias and the acute phase of chronic leukaemias. Of 35 cases with leukaemias, lactoferrin-negative polymorphonuclear leucocytes (type 0) were observed in the following cases: eight cases of acute myelogenous leukaemia (8/14), a case of chronic myelogenous leukaemia (1/10) in blast crisis, one of acute promyelocytic leukaemia (1/1), one of acute monocytic leukaemia (1/2), and a case of chronic myelomonocytic leukaemia (1/2) in a transitional phase to an acute myelomonocytic leukaemia. In two cases of acute myelogenous leukaemia, in which the majority of polymorphonuclear leucocytes were negative for lactoferrin, ultrastructural cytochemical study revealed total lack of specific granules in these polymorphonuclear leucocytes. This suggests that lactoferrin is localised in the specific granules of neutrophils as has been postulated previously by others.

Topics: Adolescent; Adult; Aged; Bone Marrow; Child; Child, Preschool; Female; Histocytochemistry; Humans; Immunoenzyme Techniques; Lactoferrin; Lactoglobulins; Leukemia; Leukemia, Myeloid, Acute; Male; Microscopy, Electron; Middle Aged; Neutrophils; Peroxidase

Myeloperoxidase (MPO) and elastase, restricted to azurophil granules of neutrophils, as well as lactoferrin, restricted to specific granules of neutrophils, were determined in plasma and serum from patients with acute myeloid leukaemia (AML). Highly sensitive radio immuno assays were developed for detection of these proteins. Serum MPO was increased in 12/35 and decreased in 2/35 patients without correlation to WBC or neutrophil counts; these levels may reflect an abnormal production by leukaemic blasts or ineffective granulopoiesis in the bone marrow. Serum elastase was increased in 6/22 patients. Serum lactoferrin was decreased in 12/25 patients without correlation to neutrophil counts probably reflecting abnormal production. Serum elastase and MPO showed a covariation in chronic myeloid leukaemia but not in AML; the latter finding may indicate that the synthesis of these two proteins is not synchronized in AML-cells. Sequential studies of patients with AML demonstrated fluctuations of serum MPO and lactoferrin during remission most likely because of chemotherapeutic pertubation. Although a limited number of patients has been studied it is suggested that serum lactoferrin may be of help for prediction of relapse in AML.

Topics: Antineoplastic Agents; Humans; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Neutrophils; Pancreatic Elastase; Peroxidase; Peroxidases; Radioimmunoassay

Activity was seen in the breasts of a patient with galactorrhea 72 h after intravenous injection of Ga-67 citrate. Differential protein separation of breast secretion, extracted from the breast, revealed that the Ga-67 was contained primarily in the lactoferrin-rich protein fraction. Additional studies on partially purified lactoferrin revealed that lactoferrin binds Ga-67 more avidly than does transferrin. Since lactoferrin is present in high concentration non only in human colostrum and milk, but also in neutrophilic leukocytes, bone marrow, spleen, colon, tears, and in genital, salivary, and nasopharyngeal secretions, binding of Ga-67 to lactoferrin may explain the localization of Ga-67 in certain normal tissues and inflammatory lesions.

Topics: Adult; Female; Galactorrhea; Gallium Radioisotopes; Humans; Lactation Disorders; Lactoferrin; Lactoglobulins; Leukemia, Myeloid, Acute; Neoplasm Metastasis; Pregnancy; Protein Binding; Radionuclide Imaging; Spinal Cord Neoplasms

Samples from 49 cases of myeloproliferative diseases were tested by an immunocytochemical technique for leucocyte lysozyme and lactoferrin. The presence of these constituents in myeloid precursors from cases of acute and chronic myeloid leukaemia reflected the degree of cellular maturation, lysozyme appearing (as it does in normal myeloid cells) at the stage of primary granule production (in promyelocytes), while lactoferrin wad detectable only in more mature, secondary granule-containing myeloid cells. Auer rods stained positively for lysozyme, in keeping with their relationship to primary granules. Monocytes from five cases of leukaemia showing predominantly monocytic differentiation were indistinguishable from normal monocytes in their staining reactions for lysozyme despite the presence of raised serum and urinary lysozyme levels. In four cases of acute myeloid leukaemia circulating polymorphs deficient in lactoferrin were detected: in one of these cases a similar percentage of polymorphs was lysozyme negative.

Topics: Humans; Immunoenzyme Techniques; Intracellular Fluid; Lactoferrin; Lactoglobulins; Leukemia, Monocytic, Acute; Leukemia, Myeloid, Acute; Leukocytes; Monocytes; Muramidase; Myeloproliferative Disorders; Neutrophils

A solid-phase radioimmunoassay is described for measuring lactoferrin levels in normal human plasma. The sensitivity of the assay was 6 ng. per milliliter with an intraassay coefficient of variation of 4 per cent and an interassay value of 9 per cent. Healthy adult males had a mean plasma level of 1.62 mug per milliliter which was significantly higher than adult females, 1.07 mug per milliliter. Postmenopausal females had levels similar to men, 1.74 mug per milliliter, while younger women had a significantly lower mean value, 0.75 mug per milliliter. Two menstruating women and 2 pregnant women had moderately elevated levels. Consistently elevated levels were found in patients with untreated or relapsing chronic myeloid leukemia--all over 12.0 mug per milliliter, while patients on marrow suppressant therapy tended to have subnormal levels. The collection of serum specimens as opposed to plasma, resulted in inconsistently elevated levels: EDTA was the anticoagulant of choice, as heparin interfered in the radioimmunoassay system.

Topics: Adult; Age Factors; Blood Specimen Collection; Female; Humans; Immune Sera; Iodine Radioisotopes; Lactoferrin; Lactoglobulins; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Male; Menopause; Menstruation; Middle Aged; Pregnancy; Radioimmunoassay; Sex Factors