lactoferrin and Leukemia

Topics: Collagenases; Cytoplasmic Granules; Gene Expression Regulation; Hematopoiesis; Humans; Lactoferrin; Leukemia; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Neutrophils; Transcobalamins

The role of iron in certain clinical infections is revealed. In normal persons the antibacterial and antifungal properties of blood and other tissue fluids cannot be maintained unless there are exceptionally low levels of available iron. This is controlled by the presence of the unsaturated iron-binding proteins, transferrin and lactoferrin. In several clinical conditions an abnormal availability of iron is responsible for fatal septicaemia. This is because the phagocytic system is overwhelmed by rapidly growing organisms when iron is freely available.

Topics: Bacterial Infections; Candidiasis; Carrier Proteins; Disease Susceptibility; Humans; Iron; Iron-Binding Proteins; Lactoferrin; Leukemia; Transferrin; Transferrin-Binding Proteins; Virulence

Topics: Animals; Ferritins; Hematopoiesis; Hematopoietic Stem Cells; Histocompatibility Antigens; HLA Antigens; Humans; Interferons; Lactoferrin; Leukemia; Mice; Prostaglandins E; Transferrin

Topics: Colony-Stimulating Factors; Granulocytes; Hematopoiesis; Humans; Lactoferrin; Lactoglobulins; Leukemia; Macrophages; Oncogenes; Transferrin

It is apparent from the above that molecules can have more than one role, but these roles need not be mutually exclusive. A clear understanding of cell regulations will require knowledge of all interacting molecules and the cells producing and responding to these molecules. This will be especially important when studies on the roles of these molecules in maintenance of long-term marrow and blood cultures are investigated further.

Topics: Animals; Cells, Cultured; Colony-Stimulating Factors; Depression, Chemical; Ferritins; Growth Inhibitors; Hematopoiesis; Hematopoietic Stem Cells; Histocompatibility Antigens Class II; Humans; Hydrogen-Ion Concentration; Interferon Type I; Interferon-gamma; Lactoferrin; Leukemia; Leukemia, Experimental; Leukocytes; Mice; Models, Biological; Monocytes; Prostaglandins E; Transferrin

The transferrins are iron-binding proteins with molecular weights of around 80,000, which interact with a maximum of two ferric atoms per each protein molecule. The best known transferrins are the serotransferrins from animal sera, lactoferrins from milk, and conalbumin from egg-white. The iron-deficient transferrins will inhibit the growth of certain bacteria and fungi by making iron unavailable for bacterial metabolism. Such activity is abolished if the transferrin is saturated with iron. Many organisms can produce small molecular-weight iron-binding compounds called siderophores that can successfully utilize the iron sequestered by the transferrins. Such organisms are very virulent. Overwhelming evidence is now available to indicate that the transferrins play an important role in mammalian host-defense mechanisms. Thus, iron injections into animals infected with virulent bacteria result in increased death rates, and parenteral iron administration to human infants predisposes them to fatal septicemia. On the other hand, in cases of systemic infection, the organism responds by lowering its total serum iron, so as to make the serotransferrin present less saturated with iron. This phenomenon is called nutritional immunity. The iron apparently moves into the storage tissues from the circulation, and furthermore, it is withheld from circulation by the reticuloendothelial system. Laboratory results in such cases indicate low total serum iron levels and high unsaturated iron-binding activity values, thus increasing the bacteriostatic effects of the serotransferrins. Increased lactoferrin levels are observed in the milks of mastitic cattle.

Topics: Anemia, Hypochromic; Animals; Bacteria; Bacterial Infections; Carrier Proteins; Conalbumin; Female; Fungi; Humans; Hydroxamic Acids; Immunity, Innate; Iron; Iron Chelating Agents; Iron-Binding Proteins; Lactoferrin; Leukemia; Milk, Human; Mycoses; Pregnancy; Siderophores; Transferrin; Transferrin-Binding Proteins

Topics: Humans; Lactoferrin; Leukemia; Leukocyte Count; Neutrophils; Peroxidase

LF11-322 (PFWRIRIRR-NH2) (PFR peptide), a nine amino acid-residue peptide fragment derived from human lactoferricin, possesses potent cytotoxicity against bacteria. We report here the discovery and characterization of its antitumor activity in leukemia cells. PFR peptide inhibited the proliferation of MEL and HL-60 leukemia cells by inducing cell death in the absence of the classical features of apoptosis, including chromatin condensation, Annexin V staining, Caspase activation and increase of abundance of pro-apoptotic proteins. Instead, necrotic cell death as evidenced by increasing intracellular PI staining and LDH release, inducing membrane disruption and up-regulating intracellular calcium level, was observed following PFR peptide treatment. In addition to necrotic cell death, PFR peptide also induced G0/G1 cell cycle arrest. Moreover, PFR peptide exhibited favorable antitumor activity and tolerability in vivo. These findings thus provide a new clue of antimicrobial peptides as a potential novel therapy for leukemia.

Topics: Animals; Antimicrobial Cationic Peptides; Antineoplastic Agents; Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; Female; Hemolysis; HL-60 Cells; Humans; Lactoferrin; Leukemia; Mice; Necrosis; Peptides

Cationic antimicrobial peptides such as bovine lactoferricin (LfcinB) constitute an important innate defense mechanism against many microbial pathogens. LfcinB also binds to and selectively kills human cancer cells via a mechanism that involves reactive oxygen species (ROS) generation and caspase activation. The antimicrobial core of LfcinB consists of only six amino acids (RRWQWR), referred to in this study as LfcinB6. Although free LfcinB6 is devoid of cytotoxic activity against cancer cells, we show here that adding a cell-penetrating hepta-arginine sequence via a glycine-glycine linker to LfcinB6 generates a peptide (MPLfcinB6) that is selectively cytotoxic for human T-leukemia and B-lymphoma cells. Flow cytometric analysis of propidium iodide and fluorescein isothiocyanate-dextran uptake by MPLfcinB6-treated cancer cells revealed extensive damage to the cell membrane, which was confirmed by scanning electron microscopy. MPLfcinB6-induced cytotoxicity was also associated with sequential ROS production and mitochondrial membrane permeabilization; however, neither ROS nor caspase activation caused by the loss of mitochondrial membrane integrity was essential for peptide-mediated cell death. We conclude that MPLfcinB6 selectively kills human T-leukemia and B-lymphoma cells by causing extensive and irreparable damage to the cell membrane.

Topics: Amino Acid Sequence; Animals; Antimicrobial Cationic Peptides; Cattle; Cell Line, Tumor; Cell Survival; Flow Cytometry; Humans; Lactoferrin; Leukemia; Lymphoma, B-Cell; Membrane Potential, Mitochondrial; Microscopy, Electron, Scanning; Reactive Oxygen Species

The objectives of this study were to evaluate the local oral defense mechanisms during the course of leukemia, and to define the correlation between the activity of salivary antibacterial factors and the oral clinical findings.. A total of 44 children with newly diagnosed acute leukemia participated in the study. The control group consisted of 23 healthy children. The examination took place at the time of the diagnosis, and during and at the end of the chemotherapy treatment course. During the collection of resting mixed saliva samples the salivary flow rate was measured. In the saliva's supernatant the following parameters were determined: total protein, peroxidase, myeloperoxidase, lysozyme, lactoferrin, and secretory immunoglobulin A.. The introduction of chemotherapy caused a slight decrease of salivary secretion rate (P < .05), as well as the decrease of S-IgA concentration (P < .01), which remained at the same level after the end of chemotherapy (P < .001). Patients with aplasia had decreased levels of peroxidase (P = .014) and myeloperoxidase (P = .013). Patients with oral mucositis presented with lower myeloperoxidase (P = .026) and peroxidase (P = .003) activity levels as well as the drop of S-IgA (P = .000) concentration compared with subjects with no mucositis.. Antileukemic treatment contributes to the compromise of salivary defense mechanisms, therefore it is reasonable to support pharmacologically the saliva's antibacterial potential of leukemic patients to impede the development of local infection.

Topics: Adolescent; Anti-Infective Agents; Antineoplastic Agents; Antioxidants; Child; Child, Preschool; Female; Humans; Immunoglobulin A, Secretory; Lactoferrin; Leukemia; Male; Mucositis; Muramidase; Opportunistic Infections; Peroxidase; Saliva; Salivary Proteins and Peptides; Secretory Rate; Stomatitis

Model studies have shown that peptides derived from the N-terminal region of bovine lactoferrin (Lf-B) exhibit antitumor activity against certain cell lines. This activity is due primarily to the peptides' apoptogenic effect. Several reports indicate that cationic residues clustered in two regions of the peptide sequence can be shuffled into one region and thereby increase cytotoxic activity, although the mechanism of this enhanced cytotoxic effect has not been clarified. In this paper, we considered several parameters that determine the mode of cell death after exposure to a native Lf-B derived peptide (Pep1, residues 17-34), and a modified peptide (mPep1) wherein the cationic residues of Pep1 are clustered in a single region of its helical structure. We found that the cytotoxic activity of mPep1 was about 9.6 fold-higher than that of Pep1 against HL-60 cells, as determined by the 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2 H-tetrazolium (MTS) assay. In investigating the expression of phosphatidylserine, we observed that the native peptide (Pep1) caused both apoptotic cell death and necrotic cell death, depending on the concentration of the peptide. In contrast, the action of mPep1 was exclusively characteristic of necrotic cell death. This observation was further confirmed by agarose gel electrophoresis, in which clear ladder-like DNA bands were observed from cells exposed to Pep1, whereas DNA from cells treated with mPep1 produced a smeared pattern. We extended the study by investigating the release of mitochondrial cytochrome c into the cytosol, and the activation of caspase-3; both peptides caused the release of cytochrome c into the cytosol, and the activation of caspase-3.These results suggest that Pep1 may kill cancer cells by activating an apoptosis-inducing pathway, whereas mPep1 causes necrotic cell death by destroying cellular membrane structure notwithstanding sharing some cellular events with apoptotic cell death.

Topics: Amino Acid Sequence; Apoptosis; Blotting, Western; Caspase 3; Cell Survival; Cytochromes c; Flow Cytometry; HL-60 Cells; Humans; Lactoferrin; Leukemia; Molecular Sequence Data; Necrosis; Peptides

Bovine lactoferricin (LfcinB) is a cationic, amphipathic peptide that is cytotoxic for human and rodent cancer cells. However, the mechanism by which LfcinB causes the death of cancer cells is not well understood. Here, we show that in vitro treatment with LfcinB rapidly induced apoptosis in several different human leukemia and carcinoma cell lines as determined by DNA fragmentation assays and phosphatidylserine headgroup inversion detected by Annexin V binding to the surface of cancer cells. Importantly, LfcinB treatment did not adversely affect the viability of untransformed human lymphocytes, fibroblasts, or endothelial cells. Studies with different LfcinB-derived peptide fragments revealed that the cytotoxic activity of LfcinB resided within the amino acid sequence FKCRRWQWRM. Treatment of Jurkat T leukemia cells with LfcinB resulted in the production of reactive oxygen species followed by caspase-2-induced dissipation of mitochondrial transmembrane potential and subsequent activation of caspase-9 and caspase-3. Selective inhibitors of caspase-2 (Z-VDVAD-FMK), caspase-9 (Z-LEHD-FMK), and caspase-3 (Z-DEVD-FMK) protected both leukemia and carcinoma cells from LfcinB-induced apoptosis. Conversely, a caspase-8 inhibitor (Z-IETD-FMK) had no effect, which argued against a role for caspase-8 and was consistent with the finding that death receptors were not involved in LfcinB-induced apoptosis. Furthermore, Jurkat T leukemia cells that overexpressed Bcl-2 were less sensitive to LfcinB-induced apoptosis, which was characterized by mitochondrial swelling and the release of cytochrome c from mitochondria into the cytosolic compartment. We conclude that LfcinB kills cancer cells by triggering the mitochondrial pathway of apoptosis at least in part through the generation of reactive oxygen species.

Topics: Animals; Annexin A5; Anti-Bacterial Agents; Antioxidants; Apoptosis; Carcinoma; Caspase 2; Caspase 3; Caspase 9; Caspases; Cattle; Cell Line, Tumor; Cell Survival; DNA Fragmentation; Dose-Response Relationship, Drug; Endothelial Cells; Enzyme Activation; Enzyme Inhibitors; Fibroblasts; Humans; Immunoblotting; Jurkat Cells; Lactoferrin; Leukemia; Lymphocytes; Membrane Potentials; Microscopy, Electron; Mitochondria; Oligopeptides; Phosphatidylserines; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles; Time Factors

We previously demonstrated that lactoferrin gene polymorphisms occur in cancer cells of patients with leukemia and breast cancer. In this study, we established a non-radioactive polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, one of the most sensitive and simplest methods to detect polymorphisms and mutations of the human lactoferrin gene. We optimized the PCR conditions for nine different DNA templates and 16 pairs of exon primers for SSCP analysis. The DNA templates used in the analyses were prepared from a cosmid clone (CT6-1) that contains the human lactoferrin gene, human placental tissue, leukocytes from 10 normal volunteers, leukemic cells of two patients, and previously established three breast and two leukemic cell lines. With the appropriate exon-primer sets, PCR products from exon I to exon 16 of the lactoferrin gene were generated from the DNA templates and analyzed by SSCP. Compared with the homogenous cloned DNA, lactoferrin gene polymorphisms were detected within exons 2, 5, 7, 9, 13, 14, and 15 of the normal placental and leukocyte DNA. In addition, abnormal migration patterns of the lactoferrin gene in cancer cells were detected in exons 4, 5, 13, 14, and 15. The PCR-SSCP band migration patterns can be attributed either to gene polymorphism in normal cells or to DNA mutations in cancer cells and the employed method cannot distinguish between them. Nonetheless, the present analysis suggests that genetic polymorphisms of the lactoferrin gene exist in selected exons and additional mutations of the lactoferrin gene do occur in the cancer cells.

Topics: Breast Neoplasms; DNA Mutational Analysis; Exons; Genetic Testing; Humans; Lactoferrin; Leukemia; Placenta; Polymerase Chain Reaction; Polymorphism, Genetic; Polymorphism, Single-Stranded Conformational; Tumor Cells, Cultured

The lactoferrin gene is highly expressed in many different tissues, and its expression is controlled by different regulators. In this report, we have defined a retinoic acid response element (RARE) in the 5'-flanking region of the lactoferrin gene promoter. The lactoferrin-RARE is composed of two AGGTCA-like motifs arranged as a direct repeat with 1-bp spacing (DR-1). A gel retardation assay demonstrated that it bound strongly with retinoid X receptor (RXR) homodimers and RXR-retinoic acid receptor (RAR) heterodimers as well as chicken ovalbumin upstream promoter transcription factor (COUP-TF) orphan receptor. In CV-1 cells, the lactoferrin-RARE linked with a heterologous thymidine kinase promoter was strongly activated by RXR homodimers in response to 9-cis-retinoic acid (9-cis-RA) but not to all-trans-RA. When the COUP-TF orphan receptor was cotransfected, the 9-cis-RA-induced RXR homodimer activity was strongly repressed. A unique feature of the lactoferrin-RARE is that it has an AGGTCA-like motif in common with an estrogen-responsive element (ERE). The composite RARE/ERE contributes to the functional interaction between retinoid receptors and the estrogen receptor (ER) and their ligands. In CV-1 cells, cotransfection of the retinoid and estrogen receptors led to mutual inhibition of the other's activity, while an RA-dependent inhibition of ER activity was observed in breast cancer cells. Furthermore, the lactoferrin-RARE/ERE showed differential transactivation activity in different cell types. RAs could activate the lactoferrin-RARE/ERE in human leukemia HL-60 cells and U937 cells but not in human breast cancer cells. By gel retardation analyses, we demonstrated that strong binding of the endogenous COUP-TF in breast cancer cells to the composite element contributed to diminished RA response in these cells. Thus, the lactoferrin-RARE/ERE functions as a signaling switch module that mediates multihormonal responsiveness in the regulation of lactoferrin gene expression.

Topics: Base Sequence; COUP Transcription Factor I; DNA-Binding Proteins; Estrogens; Female; Gene Expression Regulation; Humans; Lactoferrin; Leukemia; Molecular Sequence Data; Promoter Regions, Genetic; Protein Binding; Receptors, Retinoic Acid; Recombinant Proteins; Retinoid X Receptors; Signal Transduction; Transcription Factors; Transcription, Genetic; Tretinoin; Tumor Cells, Cultured

New technology allows highly sensitive flow cytometric detection and quantitative analysis of intracellular antigens in normal and malignant hemopoietic cells. With this technology, the earliest stages of myeloid and lymphoid differentiation can easily and reliably be identified using antibodies directed against (pro-)myeloperoxidase/MPO, CD22 and CD3 antigens, respectively. Particularly for the analysis of undifferentiated acute myeloblastic leukemia (AML) cells, the immunological demonstration of intracellular MPO or its enzymatically inactive proforms is highly relevant, since other myeloid marker molecules such as CD33, CD13, or CDw65 are either not restricted to the granulomonocytic lineage or appear later in differentiation. By combining MPO staining with staining for lactoferrin (LF), undifferentiated cells can be distinguished from the granulomonocytic maturation compartment in bone marrow, since LF is selectively expressed from the myelocyte stage of differentiation onward. The list of informative intracellular antigens to be used in leukemia cell analysis will certainly expand in the near future. One candidate, intracellular CD68, has already been tested by us, and results are presented. Also dealt within this article are surface marker molecules not (as yet) widely used in leukemia cell analysis but with the potential to provide important additional information. Among them are the surface structures CD15, CD15s, CDw65, CD79a (MB-1), CD79b (B29), CD87 (uPA-R), and CD117 (c-kit).

Topics: Antigens, CD; Antigens, CD34; Antigens, Differentiation, B-Lymphocyte; Antigens, Differentiation, Myelomonocytic; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; Antigens, Surface; Biomarkers, Tumor; CD3 Complex; CD79 Antigens; Cell Adhesion Molecules; Flow Cytometry; Humans; Immunophenotyping; Lactoferrin; Lectins; Leukemia; Lewis X Antigen; Membrane Glycoproteins; Peroxidase; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-kit; Receptor Protein-Tyrosine Kinases; Receptors, Antigen, B-Cell; Receptors, Cell Surface; Receptors, Colony-Stimulating Factor; Receptors, Urokinase Plasminogen Activator; Sialic Acid Binding Ig-like Lectin 2

Lactotransferrin (Lf), an iron-binding glycoprotein present as a major component in the specific granules of human neutrophilic granulocytes is released in the blood during the acute phase of infection and participates in the regulation of the host-defence mechanisms. Our previous observations (Mazurier et al., 1989) showing i) that the activation by PHA of T-lymphocytes induces the appearance at the cell surface of Lf-receptors which are absent from the membrane of resting lymphocytes and ii) that Lf becomes a growth factor for the activated lymphocytes, led us to undertake a series of researches on the presence of Lf receptors at the surface of different blood cells. Characterization of Lf receptors was performed by flow cytofluorimetry using either Lf labelled on its glycan moiety with fluorescein or purified anti-lymphocyte Lf receptor antibodies. High affinity receptors for Lf were characterized only at the surface of human activated lymphocytes and of non-activated platelets. These two receptors possess common physicochemical properties and antigenic epitopes. Low affinity receptors for Lf were characterized on monocytes, eosinophils and neutrophils. These receptors are immunologically different from those found on activated lymphocytes and on non-activated platelets. Cell-lines of human lymphocyte T and megakaryocyte possess lactotransferrin receptors whose properties are similar to those found on peripheral blood cells. The soluble form of the receptor identified in the lymphocytes T culture medium possesses a molecular mass close to that of the membrane receptor suggesting that the cytoplasmic tail of the receptor should be very short.

Topics: Blood Platelets; Eosinophils; Flow Cytometry; Humans; Lactoferrin; Leukemia; Leukocytes; Lymphocytes; Monocytes; Receptors, Cell Surface; Tumor Cells, Cultured

Topics: Antigens, CD; Antigens, Differentiation, B-Lymphocyte; Bone Marrow; Bone Marrow Cells; CD3 Complex; Cell Adhesion Molecules; Flow Cytometry; Humans; Immunophenotyping; Lactoferrin; Lectins; Leukemia; Peroxidase; Sialic Acid Binding Ig-like Lectin 2

Human lactoferrin has been found to be decreased or absent in most breast cancer and leukemia cells. In order to examine the lactoferrin gene for both structural alterations and the degree of methylation, we isolated a 2117-kilobase complementary DNA from human breast tissue. This complementary DNA was used to probe DNA extracted from normal peripheral blood, leukemia cells from patients, leukemia cell lines, and breast cancer cell lines. Immunocytochemical staining of these cells confirmed the decreased production of lactoferrin in malignancy. MspI restriction enzyme fragment patterns demonstrated genetic polymorphism which occurred in DNA from both normal and malignant cells. Polymorphism was also noted with XbaI. In this case, there were two fragment patterns that were only found in DNA from malignant cells. The degree of DNA methylation was also evaluated. The methylation pattern of DNA extracted from malignant cells was highly variable and generally less methylated than DNA extracted from normal WBCs. It is possible that the decrease in lactoferrin associated with cancer is multifactorial and includes gene structural changes as well as altered regulation. Further study is needed to determine whether the changes found in this study are the result of the malignancy or contribute to its onset or maintenance.

Topics: Amino Acid Sequence; Breast Neoplasms; Gene Library; Genes; Humans; Lactoferrin; Leukemia; Leukocytes; Methylation; Molecular Sequence Data; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length

We have developed a technique that permits evaluation and semi-quantification of iron-binding function in mature neutrophils. Neutrophil iron-binding reactivity (NFeBR) visualized using the iron nitrilotriacetate-acid ferrocyanide technique was rated 0 to 5+ in 100 segmented cells; the ratings were totaled to yield a score (NFeBRS). Males and post-menopausal females had significantly higher NFeBRS than pre-menopausal females. Neonates had low values, and a homogeneous distribution of NFeBR among neutrophils. In pregnancy and acute infection, NFeBRS were significantly increased. In a patient with congenital lactoferrin (Lf) deficiency, the NFeBRS was very low. In Ph1-positive chronic myelogenous leukemia, 13 of 17 patients had low NFeBRS due to decreased NFeBR, which was heterogeneously distributed among mature neutrophils. By ultrastructural analysis of mature neutrophils in two such patients, the stain deposits in FeBR-positive granules were of normal intensity, but the numbers of positive granules were decreased in many cells. NFeBRS were also low in 12 of 23 patients with other myeloproliferative disorders, and in seven of 15 patients with acute non-lymphoblastic leukemia, but in only seven of 63 patients with other neoplasms. NFeBRS were significantly correlated (p less than 0.008) with values of neutrophil Lf content quantified by immunologic assays in a wide variety of conditions and over a broad range of values. These results augment observations of neutrophil Lf made using immunological methods.

Topics: Adult; Age Factors; Cell Compartmentation; Histocytochemistry; Humans; In Vitro Techniques; Iron; Lactoferrin; Leukemia; Myeloproliferative Disorders; Neoplasms; Neutrophils

Bone-marrow regeneration after chemo- and radiotherapy-induced aplasia can be monitored by serum levels of myeloperoxidase (MPO), lysozyme (LYS) and lactoferrin (LF). In 10 patients with leukemia, serum measurements were performed before and after bone-marrow transplantation. Bone-marrow regeneration was suggested by increments in serum MPO and LYS 5 and 4 days prior to the increase in mononuclear cells (Mono) and 10 and 9 d before the increase in polymorphonuclear leukocytes (PMN) in the peripheral blood. LF started to rise 4.5 d before detectable circulating PMNs. 2 patients with early relapses of leukemia post transplantation are shown to display atypical patterns of serum MPO and LYS. We conclude that serum measurements of MPO, LYS and LF may be used as early and sensitive means to monitor bone-marrow activity during hematological regeneration. However, the findings also strongly support the earlier proposal that MPO alone may be used to reflect myeloid activity in the bone-marrow in general.

Topics: Adolescent; Adult; Bone Marrow; Bone Marrow Transplantation; Child, Preschool; Female; Humans; Lactoferrin; Lactoglobulins; Leukemia; Leukocyte Count; Male; Muramidase; Peroxidase; Regeneration; Remission Induction

Central nervous system (CNS) involvement in patients with leukaemia or lymphoma presents a diagnostic problem. This study was conducted to test whether combined measurements of various cellular markers such as beta 2-microglobulin (beta 2m), lactoferrin (LF) and lysozyme (LYS) in the cerebrospinal fluid (CSF) might aid in the diagnosis of CNS involvement in such patients. Forty-two patients were studied. Sixteen were considered to have CNS involvement and 26 showed no signs of such involvement. In the group with symptoms or signs of CNS involvement, nine patients out of 12 had increased total protein in CSF, 14 of 14 increased beta 2m, 14 of 16 increased LYS and five of 15 increased LF. In patients without CNS involvement total protein was increased in four of 25, beta 2m in three of 21, LYS in four of 28 and LF in one of 28 patients. The differences were statistically significant (P less than 0.01, P less than 0.001, P less than 0.001 and P less than 0.05, respectively). Prophylactic intrathecal methotrexate treatment in patients with acute lymphoblastic leukaemia caused an increase in the CSF of beta 2m, LYS and LF but not of total protein, which may reflect a drug-induced inflammatory reaction in the CNS. We conclude that combined measurements of the three cell markers add to our understanding of the cellular reaction to malignant cells in the CNS in leukaemia and lymphoma and may be valuable supplements in the diagnosis of this CNS involvement.

Topics: Adolescent; Adult; Aged; beta 2-Microglobulin; Central Nervous System Diseases; Female; Humans; Injections, Spinal; Lactoferrin; Lactoglobulins; Leukemia; Lymphoma; Male; Methotrexate; Middle Aged; Muramidase; Time Factors

A solid-phase, one-step radioimmunoassay was developed for the determination of plasma lactoferrin concentration. The detection limit of the assay is 150 micrograms/l. Leakage of cellular lactoferrin was minimal when EDTA was used as anticoagulant, while results obtained from serum and from heparinized plasma were not reproducible. The plasma lactoferrin concentration of 35 female and 44 male healthy adults was measured in order to determine normal values. The geometric mean of lactoferrin levels in men is about 10% higher than in women: 483 (200-1500) micrograms/l in men and 446 (200-870) micrograms/l in women. Patients with acute and chronic leukaemias were also studied. In 38 patients with chronic myeloid leukaemia plasma lactoferrin levels were increased by three times while the neutrophil count was ten times higher than normal. Normal lactoferrin concentrations were measured in plasma samples from 15 patients with chronic lymphocytic leukaemia in incomplete remission while no detectable lactoferrin was found in samples from those in relapse (10 patients). In the untreated patients or those in relapse (19 cases) of both acute lymphocytic and myeloid leukaemias, plasma lactoferrin concentrations were undetectable while they seemed to return to normal during remission (3 cases). The data obtained indicate that the determination of plasma lactoferrin concentration might play an important role in facilitating the assessment of total blood granulocyte pool (TBGP).

Topics: Blood Cell Count; Female; Humans; Lactoferrin; Lactoglobulins; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid; Male; Neutrophils; Prognosis; Radioimmunoassay; Reference Values

Lactoferrin is a major constituent of polymorphonuclear leukocyte granules and is present in mature neutrophils but not in blasts or promyelocytes. We have isolated a cDNA probe for lactoferrin and used it to study the synthesis of lactoferrin mRNA by normal and leukemic granulocyte precursors. The probe pHL-41 has been subcloned in phage m13 and characterized by restriction endonuclease analysis and nucleic acid sequencing. pHL-41 contains approximately 40% of the coding sequence of the lactoferrin gene. The 3' untranslated region includes a stop codon and a possible polyadenylation signal. There is a greater than 98% agreement between the cDNA-deduced amino acid sequence and that determined by analysis of the protein. Myeloid cells from normal bone marrow and circulating leukocytes from patients with chronic granulocytic leukemia contain lactoferrin mRNA transcripts that are indistinguishable in size and relative quantity. The human promyelocytic leukemia cell line HL-60 contains no lactoferrin mRNA. Induction of monocytic or granulocytic differentiation fails to induce the synthesis of detectable lactoferrin message. Similarly, studies with the human myeloblastic leukemia cell line PLB-985 reveal the inability of these cells to produce lactoferrin mRNA even under conditions that bring about morphologically demonstrable granulocytic differentiation. These data suggest that granulocytic differentiation in the leukemic cell lines is incomplete or defective. The presence of lactoferrin may play a role in the orderly expression of the genetic program leading to the development of the normal mature granulocyte.

Topics: Base Sequence; Bone Marrow; Bone Marrow Cells; Cell Line; DNA; Humans; Lactoferrin; Lactoglobulins; Leukemia; Leukemia, Experimental; Leukemia, Myeloid; Molecular Sequence Data; RNA, Messenger

Plasma concentrations of lactoferrin were measured in immediately separated EDTA samples from 5 subjects who had received HLA identical bone marrow transplants for leukaemia or aplastic anaemia and from 7 subjects who were leukopenic as a consequence of chemotherapy for a variety of malignant conditions. Plasma lactoferrin concentrations were found to closely parallel the leucocyte count and were not found to either predict or to antedate leucocyte regeneration. Serial measurements of plasma lactoferrin in a subject with no circulating neutrophils who received a bone marrow graft revealed that the clearance of lactoferrin followed an exponential pattern and had an initial half time of 2.2 h.

Topics: Agranulocytosis; Anemia, Aplastic; Bone Marrow Transplantation; Hematopoiesis; Humans; Kinetics; Lactoferrin; Lactoglobulins; Leukemia; Leukocyte Count; Neoplasms; Neutropenia; Neutrophils

Serum levels of lactoferrin, lysozyme and myeloperoxidase have been established in 31 healthy children. On average, serum lactoferrin was 330 micrograms/1, serum lysozyme 1638 micrograms/1 and serum myeloperoxidase 174 micrograms/1. Serum myeloperoxidase was, on average, significantly higher in children than in adults (p = 0.01), whereas serum lactoferrin and serum lysozyme were equal to those of adults. In a group of infection-prone children (n = 31), both serum lactoferrin and serum myeloperoxidase, but not the serum lysozyme levels, were significantly lower (p less than 0.001 and p = 0.002, respectively) than those of the reference children in spite of normal intracellular contents and even somewhat higher peripheral blood polymorphonuclear counts. Based on the assumption that serum lactoferrin and serum myeloperoxidase reflect turnover and activity of neutrophil granulocytes, the findings could suggest reduction in these respects and could be one contributing factor to the high infection propensity of these children. Serum levels of the three proteins have also been measured in 10 children with suspected or various forms of manifest leukemia. It is suggested that the levels reflect turnover and stage of maturation of the myeloid and monocytic cells and could, therefore, aid in the understanding and diagnosis of these diseases.

Topics: Adolescent; Adult; Bacterial Infections; Cell Division; Child; Child, Preschool; Female; Humans; Infant; Lactoferrin; Lactoglobulins; Leukemia; Leukocyte Count; Leukocytes; Male; Muramidase; Peroxidase; Peroxidases; Probability

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

A serum-free medium supplemented with growth factor(s) was devised to grow human lymphocytic cell lines. The medium was developed using human lymphocytic cell line, Bri 7 cells. In the process of constructing the medium, human lactoferrin was found to be an essential growth factor for the cell line. Human lactoferrin has higher growth stimulatory activity than human transferrin, and was sensitive to heat. Long-term cultivation of the cells was achieved in the defined medium supplemented with human lactoferrin only. The defined medium specifically supported the growth of various other human B- and T-lymphocytic cell lines but not the growth of various mouse lymphocytic cell lines. In lactoferrin-supplemented medium, the growth of some human cell lines were further stimulated by the addition of a combination of insulin, ethanolamine and selenium, or another combination of 2-mercaptoethanol and the above three factors. Bovine lactoferrin could be substituted for human lactoferrin.

Topics: Animals; Cattle; Cell Division; Cell Line; Female; Growth Substances; Humans; Kinetics; Lactoferrin; Lactoglobulins; Leukemia; Lymphocytes; Mice; Milk; Milk, Human; Pregnancy

The concentration of lactoferrin, a non-heme iron binding glycoprotein, was determined in more than 1500 EDTA plasma samples by a 2-site solid phase immunoradiometric assay to assess the significance of lactoferrin in plasma and to investigate applications for this assay. The use of commercially available antibody and antigen and a relatively short assay time make this assay more suitable for use in routine clinical laboratories than previous methods. A normal range of 250-750 micrograms/l was established. There was a correlation between plasma lactoferrin concentration and the circulating blood neutrophil count in most patients except those with splenomegaly, post-splenectomy and undergoing intensive chemotherapy. Patients with gross splenomegaly usually had an increased and post-splenectomy patients a decreased lactoferrin/neutrophil ratio indicating a respective increase and decrease in the marginated pool. In patients with acute leukemia after chemotherapy or transplantation plasma lactoferrin levels increased 1 to 5 d before blood neutrophil counts rose. As plasma lactoferrin seems to be derived from neutrophils, its concentration is probably related to the size of the total blood granulocyte pool. Calculation of the lactoferrin/neutrophil ratio demonstrated variations in the size of the bone marrow reserve and the marginated neutrophil pool.

Topics: Humans; Lactoferrin; Lactoglobulins; Leukemia; Leukocytes; Neutrophils; Postoperative Period; Radioimmunoassay; Splenectomy; Splenomegaly

In the present paper we apply the "ecotaxis hypothesis" to the analysis of lymphocyte distribution in Hodgkin's disease and other forms of lymphoid malignancy. The results lead us to consider the possiblity that metal-binding proteins, namely ferritin, transferrin and lactoferrin, play a role in lymphocyte ecotaxopahty. It is suggested that in Hodgkin's disease, a failure of lymph node and spleen monocytes to handle iron normally could explain most of the hematologic, immunologic, pathologic, and epidemiologic features of the disease.

Topics: Cell Movement; Female; Ferritins; Hodgkin Disease; Humans; Iron; Lactoferrin; Leukemia; Lymph Nodes; Lymphoma; Male; Monocytes; Phagocytosis; Receptors, Antigen, B-Cell; Rosette Formation; Spleen; Splenectomy; Splenic Neoplasms; T-Lymphocytes; Transferrin

In 31 patients, covering a wide range of blood neutrophil counts and turnover rates, the plasma concentrations of myeloperoxidase and lactoferrin have been measured with radioimmunoassays and compared to neutrophil kinetic parameters, measured with DF32P-labeled neutrophils. It was found that the plasma concentrations of both proteins correlated significantly with the total number of neutrophils in the blood (TBGP=total blood granulocyte pool) as well as with the neutrophil turnover rate (GTR=granulocyte turnover rate), which is evidence that neutrophilic granulocytes are the main suppliers of myeloperoxidase and lactoferrin to the plasma. In contrast to the previously demonstrated better relationship between the GTR and plasma lysozyme, a protein also originating in neutrophil granules, both myeloperoxidase and lactoferrin correlated better with the TBGP. These differences may reflect differences in the mode of release of intragranular proteins from neutrophils to the plasma. The correlation of the plasma lactoferrin concentration with the TBGP was so good as to suggest its use in the clinical assessment of the TBGP.

Topics: Arthritis, Rheumatoid; Granulocytes; Hematologic Diseases; Hodgkin Disease; Humans; Lactoferrin; Lactoglobulins; Leukemia; Leukocyte Count; Liver Cirrhosis; Lymphoma, Non-Hodgkin; Neutrophils; Peroxidase; Peroxidases