muramidase and Cell-Transformation--Neoplastic

We report a case of acute monocytic leukemia with normal serum lysozyme but with immunoperoxidase positivity for lysozyme in the leukemic blasts. The diagnosis of acute monocytic leukemia was made on the basis of cellular morphology, immunocytochemistry and cytochemistry. Discrepancy between serum lysozyme and intracytoplasmic lysozyme has not previously been reported in the literature. Levels of serum lysozyme may be misleading in cases of acute monocytic leukemia (FAB classification M5).

Topics: Adult; Cell Transformation, Neoplastic; Humans; Leukemia, Monocytic, Acute; Male; Monocytes; Muramidase; Staining and Labeling

Recent advances in analysis of leukemic cell phenotypes using cell surface markers have provided important insights into leukocyte differentiation and the cellular origin of leukemia. In addition to the traditional cell surface markers, i.e., surface membrane immunoglobulin and receptors for sheep erythrocytes that define B and T lymphocytes, highly specific monoclonal antibodies have been developed that discriminate various stages of human lymphocyte and granulocyte differentiation. Explorations of the detailed phenotypes of leukemic cells in relation to normal hemopoietic differentiation reveal that consistent, composite phenotypes of different subclasses of lymphoid malignancies closely mimic those of corresponding normal cells at equivalent levels of maturation. This is exemplified in lymphoma cells (chronic lymphocytic leukemia of B or T type, Sezary Syndrome, immunocytoma) that resemble mature and immunocompetent T and B cells, in T cell acute lymphoblastic leukemia (T-ALL) (equivalent to thymus cells) and in non-T ALL (corresponding to lymphoid progenitor cells in the bone marrow). The major phenotypes documented in different leukemias represent the level of maturation arrest imposed on the dominant subclone; this is determined by, but not necessarily synonymous with, the target cell and associated clonogenic cell population in the leukemia. The clinical significance of immunodiagnosis of leukemia cell types becomes best evidenced in acute leukemias. Besides the improvement of diagnosis by using objective criteria, clinically useful subclassifications became evident: five major subtypes of ALL are now recognized, including unclassified or null ALL, common ALL, pre-B-ALL, B-ALL and pre-T/T-ALL. In addition to disclosing that ALL is an heterogeneous disease, such classifications have proved to be prognostically significant. This is exemplified in 248 children and 145 adults with ALL which were analysed for cell type and clinical data. In addition to their utility in leukemia classification, monoclonal antibodies that identify leukemia associated antigens are becoming used therapeutically, e.g., to lyse residual leukemia cells from remission bone marrows removed from leukemia patients before reinfusion. New approaches to the treatment of leukemia in which the objective is to encourage maturation of leukemia cells rather than to achieve leukemia eradication, can be monitored by phenotyping the alterations of the cell surface, and cell markers may hopef

Topics: 5'-Nucleotidase; Acid Phosphatase; Adenosine Deaminase; Adolescent; Adult; Age Factors; Aged; Aneuploidy; Animals; Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; Antigens, Surface; Blood Platelets; Cell Differentiation; Cell Transformation, Neoplastic; Child; Child, Preschool; Chromosome Aberrations; DNA Nucleotidylexotransferase; Erythrocytes; Female; Granulocytes; Histocytochemistry; HLA Antigens; Humans; Immunoglobulins; Indoles; Infant; Leukemia; Leukocyte Count; Lymphoma; Male; Mice; Middle Aged; Monocytes; Muramidase; Neoplastic Stem Cells; Neprilysin; Nucleotidases; Periodic Acid-Schiff Reaction; Phenotype; Prognosis; Purine-Nucleoside Phosphorylase; Receptors, Antigen, B-Cell; Receptors, Complement; Receptors, Fc; Rosette Formation; Sex Factors; T-Lymphocytes

Topics: Animals; Calcitriol; Carcinogens; Cell Differentiation; Cell Transformation, Neoplastic; Cells, Cultured; Cricetinae; Esterases; Humans; Lyngbya Toxins; Macrophages; Muramidase; Phorbols; Tetradecanoylphorbol Acetate

Topics: Animals; Antibody Affinity; Antibody Formation; Blood Bactericidal Activity; Cell Transformation, Neoplastic; Chickens; Dose-Response Relationship, Radiation; Hematopoiesis; Hypersensitivity, Delayed; Immune Tolerance; Immunity, Cellular; Leukemia Virus, Murine; Lymphocytes; Lymphoma; Mice; Muramidase; Neutrophils; Ovalbumin; Rabbits; Radiation Chimera; Radiation, Ionizing; Rats; T-Lymphocytes; T-Lymphocytes, Regulatory; X-Rays

Topics: Animals; Binding Sites; Breast Neoplasms; Cell Aggregation; Cell Movement; Cell Transformation, Neoplastic; Female; Humans; Iatrogenic Disease; Immunity; Immunoglobulin G; Immunosuppression Therapy; Lectins; Lymphocyte Activation; Macrophages; Mammary Neoplasms, Experimental; Muramidase; Neoplasms; Protein Binding; Rats; T-Lymphocytes; Thymus Gland

Topics: Aging; Animals; Antibody Formation; Antigen-Antibody Reactions; Antigens; Autoantibodies; Binding Sites, Antibody; Cathepsins; Cell Transformation, Neoplastic; Cytotoxicity Tests, Immunologic; Endocytosis; Histocompatibility Antigens; Hydrolases; Immunity; Kidney; Leukocytes; Liver; Lymphocyte Activation; Lysosomes; Macrophages; Muramidase; Phagocytosis; Pinocytosis; Spleen

Topics: Aneuploidy; Basophils; Blood Platelets; Cell Transformation, Neoplastic; Child; Clinical Enzyme Tests; Cytogenetics; Eosinophilia; Fetal Hemoglobin; Fever; Hematologic Diseases; Humans; Leukemia, Myeloid; Leukocyte Count; Lymphatic Diseases; Muramidase; Primary Myelofibrosis; Prognosis; Skin Manifestations; Thrombocytosis; Vitamin B 12

A fraction of gastrointestinal stromal tumor (GIST) cells overexpress the platelet-derived growth factor receptor (PDGFR)A, although most overexpress KIT. It is not known if this is because these receptor tyrosine kinases have complementary oncogenic potential, or because of heterogeneity in the cellular origin of GIST. Little also is known about why Hedgehog (HH) signaling is activated in some GIST. HH binds to and inactivates the receptor protein patched homolog (PTCH).. Ptch was conditionally inactivated in mice (to achieve constitutive HH signaling) using a Cre recombinase regulated by the lysozyme M promoter. Cre-expressing cells were traced using R26R-LacZ reporter mice. Tumors were characterized by in situ hybridization, immunohistochemistry, immunoblot, and quantitative reverse-transcriptase polymerase chain reaction analyses. Cell transformation was assessed by soft agar assay.. Loss of Ptch from lysozyme M-expressing cells resulted in the development of tumors of GIST-like localization and histology; these were reduced when mice were given imatinib, a drug that targets KIT and PDGFRA. The Hh signaling pathway was activated in the tumor cells, and Pdgfrα, but not Kit, was overexpressed and activated. Lineage tracing revealed that Cre-expressing intestinal cells were Kit-negative. These cells sometimes expressed Pdgfrα and were located near Kit-positive interstitial cells of Cajal. In contrast to KIT, activation of PDGFRA increased anchorage-independent proliferation and was required for tumor formation in mice by cells with activated HH signaling.. Inactivation of Ptch in mice leads to formation of GIST-like tumors that express Pdgfrα, but not Kit. Activation of Pdgfrα signaling appears to facilitate tumorigenesis.

Topics: Animals; Benzamides; Cell Transformation, Neoplastic; Gastrointestinal Neoplasms; Gastrointestinal Stromal Tumors; Gene Expression; Genotype; Hedgehog Proteins; Humans; Imatinib Mesylate; Integrases; Intestinal Mucosa; Kruppel-Like Transcription Factors; Leiomyosarcoma; Mice; Muramidase; Nerve Tissue Proteins; Patched Receptors; Patched-1 Receptor; Piperazines; Promoter Regions, Genetic; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-kit; Pyrimidines; Receptor, Platelet-Derived Growth Factor alpha; Receptors, Cell Surface; Signal Transduction; Zinc Finger Protein GLI1; Zinc Finger Protein Gli2; Zinc Finger Protein Gli3

Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts a peptide containing 204 amino acids with a calculated molecular weight of 23,049 D. The predicted TRA1 protein is cysteine-rich and contains multiple cysteine doublets. A putative normal counterpart gene, named NOR1, was also isolated from a normal mouse kidney cDNA library and sequenced. NOR1 cDNA predicts a peptide containing 234 amino acids. The sequence of 201 amino acids from the C-terminal NOR1 was completely identical to that of TRA1, whereas the remaining N-terminal amino acids (33 amino acids) were longer than that (3 amino acids) of TRA1 and the N-terminus of NOR1 protein contained proline-rich sequence. A similarity search against current nucleotide and protein sequence databases indicated that the NOR1/TRA1 gene(s) is conserved in a wide range of eukaryotes, because apparently homologous genes were identified in Caenorhabditis elegans and Saccharomyces cerevisiae genomes. Northern blotting using TRA1-specific and NOR1-specific probes indicated that TRA1 mRNA is exclusively expressed in leukemogenic but not in nonleukemogenic Mm sublines and normal tissues and also indicated that NOR1 mRNA is expressed in normal tissues, especially in kidney, lung, liver, and bone marrow cells but not in any Mm sublines. After leukemogenic Mm-P cells were induced to differentiate into normal macrophages by sodium butyrate, the normal counterpart, NOR1, was expressed, whereas the TRA1 level decreased. Furthermore, transfection of TRA1 converted nonleukemogenic Mm-S1 cel

Topics: Amino Acid Sequence; Animals; Base Sequence; Caenorhabditis elegans; Carrier Proteins; Cell Transformation, Neoplastic; Cloning, Molecular; DNA-Binding Proteins; DNA, Complementary; Evolution, Molecular; Fungal Proteins; Gene Expression Regulation, Leukemic; Genes; Helminth Proteins; Leukemia, Myeloid, Acute; Membrane Proteins; Mice; Mice, Inbred Strains; Molecular Sequence Data; Monocytes; Muramidase; Neoplasm Proteins; Neoplasm Transplantation; Nerve Tissue Proteins; Nuclear Proteins; Organ Specificity; Phospholipid Transfer Proteins; Polymerase Chain Reaction; Proteins; Receptors, Steroid; Receptors, Thyroid Hormone; RNA, Messenger; RNA, Neoplasm; Saccharomyces cerevisiae; Sequence Alignment; Sequence Homology, Amino Acid; Species Specificity; Subtraction Technique; Transfection; Tumor Cells, Cultured

The in vitro characteristics of spontaneous thymomas from BUF/Mna rats, a strain with a high incidence of these tumors, were studied. In primary cultures, the adherent cells consisted of mononuclear macrophages, mono- and multi-nuclear epithelial cells and some fibroblastic cells on day 3. The macrophages rapidly increased in number with the formation of large multinuclear cells by day 9. A modest increase in the number and nuclearity of macrophages was also noted in adherent cultures of normal thymuses from 5-week-old BUF/Mna rats. On the other hand, in cultures of thymic cells from 1-year-old or 5-week-old ACI/NMs rats, a normal control rat strain, macrophages did not increase in number and only rarely formed multinuclear cells in adherent cell cultures. These results suggest that abnormal proliferation signal(s) to thymic macrophages and/or their progenitor cells accompanies and may be involved in the development of thymomas in BUF/Mna rats.

Topics: Animals; Cell Fusion; Cell Transformation, Neoplastic; Cells, Cultured; Immunohistochemistry; Macrophages; Male; Muramidase; Rats; Rats, Inbred ACI; Rats, Inbred BUF; Thymoma; Thymus Gland; Thymus Neoplasms; Tumor Cells, Cultured

Paneth cell-like change (PCLC) of the prostatic glandular epithelium was focally observed in one case of normal glandular epithelium, two cases of glandular and stromal hyperplasia, one case of prostatic intraepithelial neoplasia, and four cases of prostatic adenocarcinoma. The distinctive cells were characterized by bright, eosinophilic cytoplasmic granules on routine hematoxylin and eosin-stained material. The cytoplasmic granules in the benign prostatic epithelium were periodate-Schiff's procedure (PAS)-positive and diastase resistant and immunohistochemically negative for lysozyme, neuron-specific enolase, chromogranin, and serotonin. The eosinophilic granules in the prostatic intraepithelial neoplasia and adenocarcinoma cases were immunohistochemically positive for chromogranin, serotonin, and neuron-specific enolase, and negative for lysozyme. By electron microscopy the eosinophilic granules represented exocrine-like or lysosomal-like vesicles in the benign epithelium and neuro-endocrine granules in the malignant epithelium. The lesion represents a prostatic epithelial PCLC rather than a Paneth cell metaplasia. PCLC is the common histological manifestation of two different phenomena: (a) a PAS-positive and diastase-resistant eosinophilic cytoplasmic granular change in benign prostatic epithelium, and (b) endocrine differentiation with neuroendocrine granules in dysplastic and malignant prostatic epithelia. The importance of recognizing PCLC lies in its differentiation from other possible prostatic cytoplasmic inclusions.

Topics: Acid Phosphatase; Adenocarcinoma; Aged; alpha 1-Antichymotrypsin; Antigens, Neoplasm; Carcinoma in Situ; Cell Transformation, Neoplastic; Chromogranins; Cytoplasmic Granules; Epithelium; Humans; Immunohistochemistry; Male; Microscopy, Electron; Middle Aged; Muramidase; Phosphopyruvate Hydratase; Prostate; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Serotonin

Immunologic studies have demonstrated that the vast majority of hematolymphoid neoplasms previously designated as "histiocytic" are lymphoid in origin. Consequently, malignancies of macrophage lineage are considered rare by most authors; indeed, their existence is doubted by some. Herein we report two cases of malignant histiocytic neoplasms (malignancies of macrophage lineage) of the small intestine. Both patients presented in the 7th decade with symptoms related to an abdominal mass. The polypoid tumors protruded into the intestinal lumen, extended through the entire thickness of the bowel wall, and involved regional lymph nodes. Microscopically, sheets of large pleomorphic histiocytic cells infiltrated around crypts and were associated with an admixture of bizarre giant cells and inflammatory cells. Mitotic figures were easily found. Ultrastructurally, the cells lacked desmosomes and had indented or kidney-shaped nuclei and cytoplasm containing mostly lysosomes and dense lipid droplets. In both cases, paraffin section immunohistochemistry revealed reactivity of tumor cells for CD45RB (LCA), CD45RO (A6), CD68 (KP1), CD15 (LeuM1), and lysozyme. Frozen section immunohistochemistry performed in one case further supported the macrophage phenotype. Southern blot studies of this case did not reveal immunoglobulin or T-cell receptor beta chain gene rearrangements. One patient initially treated by surgery only died of disease 3 years after diagnosis. The second patient is alive and disease-free 2 years following postoperative combination chemotherapy. The diagnosis of malignant histiocytic neoplasms requires the use of a panel of immunohistochemical markers and may be supported by electron-microscopic studies.

Topics: Antigens, CD; Biomarkers, Tumor; Blotting, Southern; Cell Transformation, Neoplastic; DNA, Neoplasm; Gene Rearrangement; Humans; Immunohistochemistry; Intestinal Neoplasms; Intestine, Small; Lymph Nodes; Lymphoma, Large B-Cell, Diffuse; Macrophages; Male; Microscopy, Electron; Middle Aged; Muramidase

Pathologic features of eight cases of undifferentiated (embryonal) sarcoma of the liver (USL) in childhood were studied. Light microscopic examination showed a diffuse growth of spindle cells with occasional polygonal cells and multinucleated giant cells and also revealed focal areas of storiform pattern in four tumors, cambium layer formation in one tumor, and alveolar arrangement in one tumor. Immunohistochemical study showed positive staining of proliferating cells for suggestive histiocytic markers (A1AT in 6/6, A1ACT in 5/6, lysozyme in 4/6, and KP1 in 4/6) and for muscle markers (desmin in 4/6 and HHF35 in 3/6). Ultrastructural examination demonstrated that the individual tumors were composed of a mixture of cells having fibroblastic, histiocytoid, fibrohistiocytoid, myofibroblastic, and undifferentiated (primitive mesenchymal) morphologies. Also identified were cells with definite myoblastic morphology in three tumors: leiomyoblastic in one and rhabdomyoblastic in two. In conclusion, the tumor cells in USL show phenotypical diversity comparable to those of malignant fibrous histiocytoma with or without additional rhabdomyosarcomatous or leiomyosarcomatous differentiation.

Topics: Actins; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; alpha-Fetoproteins; Cell Transformation, Neoplastic; Desmin; Female; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Male; Membrane Glycoproteins; Mesenchymoma; Microscopy, Electron; Mucin-1; Muramidase; S100 Proteins; Vimentin

Topics: Animals; Biomarkers, Tumor; Carcinoembryonic Antigen; Cell Transformation, Neoplastic; Cytodiagnosis; Gastric Mucosa; Immunoenzyme Techniques; Intestinal Mucosa; Lactoferrin; Metaplasia; Muramidase; Serous Membrane

Normal colonic epithelial cells consist of several cell types or lineages that are thought to arise from a common stem cell precursor. Neoplastic transformation may occur at different stages in the differentiation of a colonic stem cell to produce tumors that may retain characteristic cell lineage phenotypes. In this study, immunohistochemical techniques were used to identify cell lineage-related markers in fetal, normal, hyperplastic, adenomatous, and cancerous colonic tissue. These markers consisted of secretory component (columnar cells), a purified mucin antigen (mucous or goblet cells), chromogranin A (enteroendocrine cells), lysozyme (Paneth cells), and carcinoembryonic antigen (panepithelial cell marker). Colonic neoplasms, like normal mucosa, predominantly expressed the markers of columnar and goblet cell lineages. Chromogranin A was expressed in a small population of cells in most normal and fetal colonic crypts. Chromogranin A reactive cells were found in 55% of hyperplastic polyps, 31% of adenomatous polyps, and 33% of carcinomas. Lysozyme reactivity was rare in fetal, normal, and hyperplastic specimens, but was present in 86% of adenomas and 40% of carcinomas. Of 42 primary carcinomas, 9% were "pluripotent" and expressed markers of all four cell lineages. In addition to columnar and goblet cell markers, 7% expressed both enteroendocrine and Paneth cell markers, 17% expressed enteroendocrine cell markers, and 24% expressed Paneth cell markers. Two cases (5%) lacked expression of any of the cell lineage markers. The remainder expressed only columnar and goblet cell markers. The markers used in this study appear to identify the major cell lineages of fetal and normal colonic epithelium and can be used to delineate the altered cell lineage phenotypes in premalignant and malignant colonic mucosa.

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Cell Transformation, Neoplastic; Chromogranin A; Chromogranins; Colon; Colonic Polyps; Colorectal Neoplasms; Humans; Immunoenzyme Techniques; Intestinal Mucosa; Mucins; Muramidase; Secretory Component; Stem Cells

A total of 66 cases of well differentiated adenocarcinomas of the gallbladder comprising 12 mucosal carcinomas and 54 advanced carcinomas were examined histologically and immunohistochemically for metaplastic changes in the tumor tissue and non-neoplastic mucosa adjacent to the tumor tissue in order to elucidate the histogenesis of gallbladder carcinoma. Among the various kinds of metaplastic changes in the gallbladder mucosa, the occurrence of endocrine cells and lysozyme immunoreactivity were used as markers. The 66 cases of adenocarcinoma were divided into 12 cases showing no metaplastic changes (non-metaplastic type) and 54 cases containing at least one marker of metaplastic changes (metaplastic type). The frequency of metaplastic changes was compared between mucosal carcinoma and advanced carcinoma to determine whether these metaplastic changes could be a phenotypic expression of the original tissue from which the tumor was derived or a secondary phenomenon associated with the progression of the tumor. No difference could be observed between the two. Moreover, the carcinoma of the non-metaplastic type was often surrounded by an ordinary mucosa without metaplastic changes, whereas the carcinoma of the metaplastic type was frequently surrounded by a metaplastic mucosa. Some cases among the non-metaplastic type carcinomas showed a morphological transition between the ordinary mucosa and the carcinoma or contained the residue of ordinary type adenoma within the tumor. On the other hand, 5 cases of the metaplastic type carcinoma contained adenomatous residue of the metaplastic type. These results suggest that there might be two types of adenocarcinoma, one being derived from the ordinary epithelium of the gallbladder and the other from the metaplastic epithelium.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Cell Transformation, Neoplastic; Epithelium; Female; Gallbladder Neoplasms; Humans; Immunohistochemistry; Male; Middle Aged; Mucous Membrane; Muramidase; Phenotype

Thirty-five patients who fulfilled the FAB diagnosis criteria of chronic myelomonocytic leukemia (CMML), i.e., myelodysplastic features, monocytosis over 10(9)/liter, bone marrow monocyte infiltration, blast cells less than 5% in the peripheral blood and less than 30% in the bone marrow, are analyzed. CMML appears as an entity distinct from myelodysplastic and myeloproliferative disorders. Splenomegaly, anemia, thrombocytopenia, leukocytosis with monocytes and granulocytic cells in all stages of development, increased blood and urine lysozyme levels without renal failure, and polyclonal hyperimmunoglobulinemia are its main clinical and biologic features. With conventional cytotoxic drugs (6-mercaptopurine, hydroxyurea), the prognosis of CMML appears poor (median survival 475 days). None of the clinical hematologic or biologic parameters tested had a significant effect on prognosis. As other chemotherapy trials seemed necessary, we recently administered small doses of cytosine-arabinoside (ARA-C) to six patients over several consecutive days and obtained a complete remission in four. These preliminary results must be confirmed by larger series using the diagnostic criteria proposed by the FAB cooperative group.

Topics: Aged; Bone Marrow; Cell Transformation, Neoplastic; Cytarabine; Female; Humans; Hydroxyurea; Leukemia, Myeloid; Male; Middle Aged; Monocytes; Muramidase; Prognosis

Several biological phenotypes of growth factor-dependent cell lines have been described in recent years, including those with T lymphocyte, neutrophil granulocyte, basophil/mast cell, B lymphocyte, and multipotential stem cell properties. The growth factors for each cell lineage are a subject of intense study. Continuous mouse bone marrow cultures infected with RNA type C viruses (retroviruses) produce nonadherent hematopoietic cells over a longer duration than control cultures. Marrow cultures derived from strains with spontaneously induced ecotropic endogenous retrovirus demonstrate a greater longevity than those from strains with no replicating virus. Cultures infected with murine leukemia virus also generate a greater number, compared with controls, of cloned permanent suspension cell lines dependent for growth on a 41,000-dalton glycoprotein (interleukin 3 [IL 3]). Some are multipotential with capacity for differentiation to erythroid, neutrophil, eosinophil, and basophil/mast cell types. Other cloned IL 3-dependent cell lines are committed to a single pathway. Studies with Friend spleen focus-forming virus indicate that the first effect in the marrow culture is mediated through a subset of adherent hematopoietic stem cells. Bone marrow culture-derived IL 3-dependent cell lines provide a model with which to study the role of viral genes in the control of differentiation and self-renewal capacity of hematopoietic stem cells.

Topics: Animals; B-Lymphocytes; Cell Line; Cell Transformation, Neoplastic; Colony-Forming Units Assay; Hematopoietic Stem Cells; Interleukin-3; Leukemia Virus, Murine; Leukemia, Experimental; Lymphokines; Mice; Mice, Inbred Strains; Muramidase; Polycythemia; Species Specificity; T-Lymphocytes

Topics: Animals; Cell Aggregation; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Erythroblasts; Erythrocyte Membrane; Erythrocytes; Friend murine leukemia virus; Gelatin; Mice; Microscopy, Electron, Scanning; Microvilli; Muramidase; Ovalbumin; Peptides; Polylysine; Proteins; Sarcosine; Surface Properties

Culture supernatants obtained from a radiation leukemia virus-transformed, hen egg-white lysozyme (HEL)-specific, suppressor T cell line are able, when injected into mice, to specifically suppress the anti-HEL antibody response. Suppression is observed on both primary and secondary anti-HEL antibody responses evaluated by direct and developed hemolytic plaque assays. Culture supernatants from this HEL-specific suppressor T cell line do not suppress the antibody response induced by a structurally related lysozyme, demonstrating the presence in the culture supernatant of a suppressor factor endowed with fine antigenic specificity. The suppressor factor is able to selectively suppress the anti-HEL antibody response induced by the N-terminal C-terminal peptide of the HEL molecule indicating that the fine specificity of this factor is restricted to an antigenic epitope present in this region of the HEL molecule. The suppressive activity is restricted by genes located within the H-2 complex and analysis of the suppression induced in recombinant mice demonstrates that the interaction between HEL-specific suppressor T cell factor and its cellular target requires identity in the I-J region of the H-2 complex.

Topics: Animals; Cell Transformation, Neoplastic; Epitopes; Female; H-2 Antigens; Leukemia Virus, Murine; Lymphokines; Male; Mice; Muramidase; Suppressor Factors, Immunologic; T-Lymphocytes; T-Lymphocytes, Regulatory

Clones of myeloid leukemic cells varying in their competence for induction of differentiation have been continuously grown in serum-free medium. In the medium used, which contained transferrin, the growth rates of these cells were nearly similar to those found in serum-containing medium. The clones also maintained in this medium their competence for induction of differentiation by the normal macrophage and granulocyte differentiation-induction protein MGI-2, the steroid dexamethasone, and lipopolysaccharide. In contrast to the results with these inducters, some clones continuously cultured in a serum-free medium showed a gain of inducibility by insulin and another clone a gain of inducibility by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate in low serum and serum-free medium. Induction of differentiation by these two compounds was therefore inhibited in these clones by the presence of serum. It is suggested that serum-free medium may also show the existence of other inducers of differentiation not detected in serum-containing medium and that these results are relevant to the possible therapeutic use of compounds such as insulin for the induction of normal differentiation in leukemic cells in vivo.

Topics: Animals; Cell Differentiation; Cell Transformation, Neoplastic; Clone Cells; Colony-Stimulating Factors; Culture Media; Dexamethasone; Hematopoietic Stem Cells; Insulin; Leukemia, Myeloid; Mice; Mice, Inbred Strains; Muramidase; Tetradecanoylphorbol Acetate; Transferrin

Retinoic acid and retinol induced functional and morphological differentiation of human promyelocytic leukemia cells (HL-60) into mature granulocytes, but did not induce functional or morphological differentiation of mouse myeloid leukemia cells (M1). Cellular retinoic acid-binding protein, but not retinol-binding protein, was detected on HL-60 cells. Neither binding protein could be detected on M1 cells. These results suggest that retinoic acid-binding protein may be necessary for induction by retinoids of functional and morphological differentiation of myeloid leukemia cells.

Topics: Animals; Carrier Proteins; Cell Transformation, Neoplastic; Cells, Cultured; Enzyme Induction; Humans; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Mice; Muramidase; Neoplasm Proteins; Rats; Receptors, Retinoic Acid; Retinol-Binding Proteins; Tretinoin

A stable bone-marrow-derived cell line, OK-BM, producing transforming avian acute leukemia virus OK 10, was established. The cell line has been maintained in culture for over 4 years and 405 passages. The cells grow rapidly in suspension, have a low serum requirement, display a uniform morphology and perpetually release transforming OK 10 virus. No transforming helper virus was detected by interference assay. The cells have a chromosome complement of the chicken. Characterization of the cells indicates that they are of myeloid origin and have properties characteristic of proliferating transformed macrophages.

Topics: Animals; Avian Leukosis Virus; Bone Marrow Cells; Cell Line; Cell Transformation, Neoplastic; Chickens; Female; Macrophages; Male; Muramidase; Phagocytosis; RNA-Directed DNA Polymerase; Time Factors

A cultured line of human myeloid leukemic cells has been used, to test for the ability of compounds used in chemotherapy to induce partial or complete differentiation of these leukemic cells. The compounds differed in their ability to induce specific differentiation-associated properties. Effectiveness of induction of Fc and C3 rosettes was of the order actinomycin C greater than cytosine arabinoside greater than mitomycin-C greater than adriamycin greater than bromodeoxyuridine greater than hydroxyurea. Induction of rosettes by actinomycin-D required a 8212-fold lower concentration than induction by hydroxyurea. All these compounds, except bromodeoxyuridine, induced the synthesis and secretion of lysozyme with the same order of effectiveness as for rosettes, but only actinomycin-D and to a lesser extent bromodeoxyuridine induced the formation of mature granulocytes. Vincristine induced only a small increase in lysozyme. The results indicate that actinomycin-D was the most potent inducer of differentiation in these human myeloid leukemic cells. It is suggested that pre-screening of individual patients for the most effective compounds that can induce differentiation of their myeloid leukemic cells in culture, may prove beneficial for treatment in a form of chemotherapy based on the induction of normal differentiation in leukemic cells.

Topics: Antineoplastic Agents; Bromodeoxyuridine; Cell Line; Cell Transformation, Neoplastic; Complement C3; Cytarabine; Dactinomycin; Doxorubicin; Humans; Hydroxyurea; Immunoglobulin Fc Fragments; Leukemia, Myeloid; Leukocytes; Mitomycins; Muramidase; Rosette Formation

An orang-utan (Pongo pygmaeus) suspension line, CP81, was shown to lack myeloid markers of lysozyme activity an d phagocytosis but to be positive for lymphocytic N-alkaline phosphatase activity, and to release a B-cell-tropic herpesvirus. This herpesvirus, termed Herpesvirus pongo, had 30--40% DNA homology with EBV and was present at 2-3 genome copies per CP-81 cell. Gibbon lymphocytes transformed by H. pongo, Epstein-Barr virus (EBV), and H. papio (of baboon, Papio hamadryas, origin) were found to be virus antigen-positive B cells. Gibbon lymphocytes transformed by H. pongo and EBV and transplanted to nude mice by the intracranial (IC) route (had a 75% and a 45% success rate, respectively), while transplants of similar cells transformed by H. papio were only 10% successful. None of these lines transplanted subcutaneously (SC) nor manifested a high degree of colony formation in 0.33% agarose (less than or equal to 0.5%), Gibbon lymphocytes transformed by H. pongo were hypodiploid while those transformed by EBV or H. papio were diploid. CP-81 cells themselves could be transplanted both IC (100%) and SC (70%) and showed a relatively high degree of colony formation in agarose (6.4-7.6%). B95-8 cells (marmoset, Saguinus oedipus-EBV) could be transplanted IC (66%) but not SC and had a low but significant ability to grow in agarose (1.6%). 594S (baboon, P. hamadryas-H. papio) cells could be transplanted IC (25%) but not SC, and grew to very low levels in agarose (0.1%).

Topics: Alkaline Phosphatase; Animals; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cell Transformation, Viral; DNA, Viral; Fluorescent Antibody Technique; Herpesviridae; Herpesvirus 4, Human; Hominidae; Lymphocyte Activation; Lymphocyte Transfusion; Male; Mice; Mice, Nude; Muramidase; Phagocytosis

The studies reviewed in this chapter provide further evidence that the secretion of lysosomal enzymes and other hydrolases is a constitutive function of certain cells whereas in other cells is an inducible process probably contributing to the pathology of a variety of diseases. Little is known of the mechanisms mediating the secretion of lysosomal enzymes. We have summarized evidence suggesting a role of microfilaments and microtubules in controlling enzyme release, but further studies of the biochemical mechanisms which control the activity of these subcellular structures are required. The fusion of lysosomes with the plasma membrane has been observed in several situations and the mechanisms underlying processes of this nature have been studied in lower organisms (Satir et al. 1973; Plattner 1974). Agents, such as concanavalin A, which interfere with the fusion of endosomes with lysosomes (Goldman 1974; Edelson and Cohn 1974a, b) should also be useful in determining the chemical nature of membrane components involved in the fusion process. New information on the fate of secreted acid hydrolases has been obtained from studies of the uptake of lysosomal enzymes by fibroblasts. Clearly, the mechanisms by which these cells endocytose secreted lysosomal enzymes will be a subject for detailed study in view of the important of directing enzymes and drugs into lysosomes (De Duve et al. 1974). The mechanisms by which extracellular inhibitors inactivate hydrolytic enzymes, particularly proteinases, is also being clarified (for review see Davies 1975) and this should aid in finding new ways for preventing tissue damage caused by the excessive secretion of these enzymes. Further investigation concerning the secretion of lysosomal enzymes should establish the essential physiological role which these enzymes play at both extracellular and intracellular sites. Also, a close examination of the interaction of both endogenous and exogenous stimuli of inflammation with cells resulting in the secretion of hydrolytic enzymes, will clarify the mechanisms underlying the initiation and progression of the inflammatory process in its diverse forms.

Topics: Animals; Anti-Inflammatory Agents; Antigen-Antibody Complex; Bone Resorption; Carrageenan; Cell Transformation, Neoplastic; Cell Wall; Complement System Proteins; Cytochalasin B; Dental Plaque; Glucuronidase; Humans; Hydrolases; Inflammation; Lysosomes; Macrophages; Microbial Collagenase; Muramidase; Neutrophils; Nucleotides, Cyclic; Plasminogen Activators; Pneumoconiosis; Streptococcus pyogenes

Cultured thioglycollate-stimulated peritoneal macrophages synthesize, accumulate, and continuously release high levels of plasminogen activators for at least 4 days whereas cultures of unstimulated macrophages do not; the higher specific catalytic activity of released vs. cell-associated enzyme suggests that the plasminogen activators are actively secreted. The major macrophage plasminogen activator is a serine protease of mol wt 48,000, and thus resembles the comparable enzyme released by virally transformed fibroblasts. Macrophages release a second plasminogen activator of mol wt 28,000 that is also a serine enzyme. The secretion products released by stimulated and unstimulated macrophages have been compared by SDS-polyacrylamide gel electrophoresis after chemical labeling with (3)H-DFP or biosynthetic labeling with (14)C-amino acids. These procedures show that some proteins are formed in both cultures, whereas others are uniquely secreted by each type of macrophage. The serine enzymes released by the two kinds of macrophages differ in specificity and electrophoretic mobility.

Topics: Amino Acids; Animals; Carbon Radioisotopes; Cell Transformation, Neoplastic; Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Female; Fibrinolysis; Isoflurophate; Macrophages; Mice; Molecular Weight; Muramidase; Peptide Hydrolases; Plasminogen; Serine; Thioglycolates; Tritium

Topics: Anemia, Hemolytic; Animals; Cell Division; Cell Line; Cell Nucleus; Cell Transformation, Neoplastic; Child, Preschool; Cytoplasm; Female; Fibroblasts; Humans; Liver; Mice; Microscopy, Electron; Microscopy, Electron, Scanning; Mitochondria, Liver; Muramidase; Rats; Ribosomes; Simian virus 40; Thymidine; Tritium

Topics: Acute Disease; Age Factors; Aged; Agranulocytosis; Anemia; Bone Marrow Examination; Cell Transformation, Neoplastic; Ecchymosis; Erythropoiesis; Female; Hematocrit; Humans; Iron; Leukemia; Leukemia, Monocytic, Acute; Leukemia, Myeloid; Leukocytes; Male; Middle Aged; Muramidase; Myeloproliferative Disorders; Splenomegaly; Syndrome; Thrombocytopenia

Topics: Alkaline Phosphatase; Cell Transformation, Neoplastic; Humans; Leukemia, Myeloid, Acute; Muramidase; Neutrophils; Peroxidases

The stimulation of DNA synthesis in mouse (C57BL) macrophages explanted in vitro was demonstrated after treatment with conditioned medium or infection with SV40. In the latter case, induction of SV40 T antigen was detected before TdR-(3)H incorporation. Even though all macrophages were infected (T antigen-positive), they exhibited considerable pleomorphism, accompanied by functional differences. Permanent lines of SV40-transformed macrophages were eventually established, and one clone was isolated which replicates indefinitely and has many properties of primary macrophages: high acid phosphatase and phagocytic activity, lysozyme production, and specific antigenic determinants. These cells differ from normal macrophages in that they contain the SV40 genome, can be trypsinized, and do not require conditioned medium for continued replication.

Topics: Acid Phosphatase; Animals; Antigens, Neoplasm; Antigens, Viral; Ascitic Fluid; Autoradiography; Carbon Isotopes; Cell Division; Cell Transformation, Neoplastic; Culture Media; DNA; DNA, Neoplasm; DNA, Viral; Fluorescent Antibody Technique; Immune Sera; Macrophages; Male; Mice; Muramidase; Phagocytosis; Simian virus 40; Thymidine; Tritium