methylcellulose and Leukemia

Topics: Adolescent; Adult; Aged; Carboxymethylcellulose Sodium; Child; Child, Preschool; Clinical Trials as Topic; Drug Evaluation; Female; Fever; Humans; Hypotension; Infant; Interferons; Leukemia; Male; Methylcellulose; Middle Aged; Neoplasms; Peptides; Poly I-C; Polylysine

The chimeric transcription factor E2a-Hlf is an oncoprotein associated with a subset of acute lymphoblastic leukemias of early B-lineage derivation. We employed a retroviral transduction-transplantation approach to evaluate the oncogenic effects of E2a-Hlf on murine B-cell progenitors harvested from adult bone marrow. Expression of E2a-Hlf induced short-lived clusters of primary hematopoietic cells but no long-term growth on preformed bone marrow stromal cell layers comprised of the AC6.21 cell line. Coexpression with Bcl-2, however, resulted in the sustained self-renewal of early preB-I cells that required stromal and interleukin-7 (IL-7) support for growth in vitro. Immortalized cells were unable to induce leukemias after transplantation into nonirradiated syngeneic hosts, unlike the leukemic properties and cytokine independence of preB-I cells transformed by p190(Bcr-Abl) under identical in vitro conditions. However, bone marrow cells expressing E2a-Hlf in combination with Bcl-2, but not E2a-Hlf alone, induced leukemias in irradiated recipients with long latencies, demonstrating both a requirement for suppression of apoptosis and the need for further secondary mutations in leukemia pathogenesis. Coexpression of IL-7 substituted for Bcl-2 to induce the in vitro growth of pre-B cells expressing E2a-Hlf, but leukemic conversion required additional abrogation of undefined stromal requirements and was associated with alterations in the Arf/Mdm2/p53 pathway. Thus, E2a-Hlf enhances the self-renewal of bone marrow B-cell progenitors without inciting a p53 tumor surveillance response or abrogating stromal and cytokine requirements for growth, which are nevertheless abrogated during progression to a leukemogenic phenotype.

Topics: Animals; B-Lymphocytes; Basic-Leucine Zipper Transcription Factors; Blotting, Southern; Blotting, Western; Bone Marrow Cells; Cell Division; Cell Separation; Cell Transformation, Neoplastic; Disease Progression; DNA; DNA-Binding Proteins; Flow Cytometry; Genotype; Interleukin-7; Leukemia; Methylcellulose; Mice; Mice, Inbred BALB C; Models, Genetic; Mutation; Oncogene Proteins, Fusion; Phenotype; Proto-Oncogene Proteins c-bcl-2; Retroviridae; Stem Cells; Time Factors; Transgenes; Tumor Suppressor Protein p53

2-Amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx) was synthesized by reacting 2-amino-5-methylphenol with bovine hemolysates. Because Phx is a phenoxazine derivative like actinomycin D, we examined its effects on the proliferation of the human leukemia cell lines K562, HL-60, and HAL-01. Phx inhibited proliferation and induced apoptosis in all of the leukemia cell lines we tested, in a dose-dependent manner. We further investigated the antitumor effect of this compound on HAL-01-bearing nude mice. Treatment with Phx markedly reduced the tumor growth rate in the experimental group, as compared with the control group. Moreover, Phx was found to have few adverse effects on weight loss and WBC count. In addition, we examined the effects of Phx on human normal hematopoietic progenitor cells by a clonogenic assay, and we observed less suppression of normal progenitor cells than of leukemic progenitors. These results suggest that Phx may be used to treat patients affected by different types of leukemia.

Topics: Animals; Antineoplastic Agents; Apoptosis; Body Weight; Cell Division; Dose-Response Relationship, Drug; HL-60 Cells; Humans; K562 Cells; Leukemia; Methylcellulose; Mice; Mice, Inbred BALB C; Oxazines; Time Factors; Tumor Cells, Cultured

The effects of recombinant products of granulocyte colony-stimulating factors (G-CSF), granulocyte/macrophage CSF (GM-CSF), human interleukin-3 (IL-3), and interleukin-1 (IL-1) were studied using purified target cell populations from patients undergoing peripheral blood stem cell transplantation after myeloablative therapy. Cells were subjected to combined purification procedures including negative selection with a panel of monoclonal antibodies (CD2, 3, 5, 10, and 20). The purified cells were enriched for HLA-DR+ (51% to 71%) and My-10+ (CD34; 37% to 54%) and had a plating efficiency of up to 20%. In the liquid-suspension limiting dilution clonal assay (LDA), purified progenitors responded directly to IL-3 by proliferation with single-hit kinetics. The ability of GM-CSF to support progenitor growth was inferior to that of IL-3, and the cells were virtually unresponsive when cultured with G-CSF, supporting the notion that these blood-derived progenitors belong to a primitive population of hematopoietic progenitor cells. The results obtained in simultaneous methycellulose cultures (MC) showed the same trend and provided additional information on the ability of GM-CSF and IL-3 to support erythroid progenitor growth. The combination of IL-3 plus G-CSF, but not IL-3 plus GM-CSF, resulted in a synergistic increase in colony number. IL-1 alpha increased both the size and number of colonies when added to IL-3 or G-CSF. Study of this enriched progenitor cell population in MC and LDA represents an excellent model for the investigation of myeloid and erythroid differentiation and for evaluating the influence of various cytokines on human hematopoiesis.

Topics: Antigens, CD; Antigens, CD34; Antigens, Differentiation; Cell Division; Cell Separation; Cells, Cultured; Child; Colony-Stimulating Factors; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Substances; Hematopoietic Stem Cells; HLA-DR Antigens; Humans; Interleukin-1; Interleukin-3; Kinetics; Leukemia; Lymphoma, Non-Hodgkin; Methylcellulose; Neuroblastoma; Recombinant Proteins

We have used a panel of 5 monoclonal antibodies against normal myeloid-differentiation antigens to determine retinoic acid-induced changes in cell surface antigens on ANLL bone marrow cells from 24 children at the time of diagnosis. Two of these antibodies (T5A7 and 5F1) detect antigens expressed on normal mature granulocytes and on all monocytes, respectively. The percentage of positive cells for each monoclonal antibody was determined by indirect immunofluorescence. After 5 days incubation with 1 microM RA in liquid culture, cells from 11 of 24 patients showed substantially increased expression of one or both antigens detected by T5A7 and 5F1. Leukemic bone marrow cells from these patients were also cultured in methylcellulose medium with and without 1 microM RA for one week, and cells from 16 of 24 patients showed clonal growth. Cultures from 10 of these 16 patients showed RA-induced inhibition of colony growth; of these 10 patients, cultures from six patients showed RA-induced increases in antigens associated with maturing myeloid cells. This suggests that the RA-induced inhibition of clonal growth observed with leukemic cells from these patients may be accompanied by the increased expression of maturation-associated myeloid antigens by these cells in the presence of RA.

Topics: Acute Disease; Antibodies, Monoclonal; Antigens, Surface; Bone Marrow; Cell Differentiation; Cell Survival; Child; Granulocytes; Humans; Leukemia; Methylcellulose; Tretinoin

In the present study we utilized a semisolid culture system with feeder cells and enriched media to evaluate the growth of acute leukemia associated with the 4;11 chromosomal translocation. We compared growth of t(4;11) leukemia to typical acute nonlymphocytic leukemia (ANL) and acute lymphocytic leukemia (ALL). The two cases of t(4;11) leukemia tested exhibited the highest cloning efficiency of cells tested. The growth characteristics of t(4;11) leukemia were more similar to ANL than ALL.

Topics: Acute Disease; Adolescent; Adult; Bone Marrow; Cells, Cultured; Child; Child, Preschool; Chromosomes, Human, 4-5; Chromosomes, Human, 6-12 and X; Colony-Forming Units Assay; Female; Humans; Infant; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Male; Methylcellulose; Monocytes; Phenotype; Recurrence; Staining and Labeling; Translocation, Genetic

Therapeutic efficacy and toxicity were evaluated in 28 children with acute lymphoblastic leukemia, in ten with acute nonlymphoblastic leukemia (ANLL), and in 13 with metastatic neuroblastoma. All were refractory to standard chemotherapeutic agents and 25 were refractory to an investigational drug. The initial dose was 12 mg/m2/day and was based on an established maximal dose tolerated in adults. This dose was found to be intolerable in 5 of 5 children with leukemia. Similarly an initial dose of 9 mg/m2/day was intolerable in 4 of 5 patients with leukemia. The starting dose in the next 28 children with leukemia or neuroblastoma was 3 mg/m2. This drug was gradually increased to the highest tolerated dose by 3-mg/m2 increments. Fifteen children with acute lymphoblastic leukemia, 3 children with ANLL, and 2 children with neuroblastoma received the drug daily. Seven patients with ANLL and 7 patients with neuroblastoma received the drug biweekly. Seventeen patients with acute lymphoblastic leukemia, 6 patients with ANLL, and 5 patients with neuroblastoma had an adequate trial of the drug. An adequate trial was defined as a minimum of 5 weeks of therapy unless progressive disease developed. Side effects of the drug were striking and included fever, hypotension, myalgia, bone pain, arthralgia, arthritis, abdominal pain, liver toxicity, thrombocytopenia, and neurotoxicity. No complete remission occurred although interferon levels above 100 units/ml were induced in nearly 50% of the patients.

Topics: Acute Disease; Adolescent; Carboxymethylcellulose Sodium; Child; Child, Preschool; Drug Evaluation; Humans; Interferon Inducers; Interferons; Leukemia; Methylcellulose; Neuroblastoma; Poly I-C; Polylysine

A Phase II study of poly(I,C)-LC was performed in 28 children and adolescents with acute lymphoblastic leukemia (ALL), 10 with acute nonlymphoblastic leukemia (ANLL), and 13 with metastatic neuroblastoma. All were refractory to standard chemotherapeutic agents and 25 to an investigational drug. Initial doses of 12 mg/m2 and 9 mg/m2 were intolerable. However, 9 mg/m2 was tolerable in the majority of patients when the drug was started at 3 mg/m2 and increased by 3 mg/m2 increments. Fifteen children with ALL, three with ANLL, and two with neuroblastoma received the drug daily. Seven patients with ANLL and seven children with neuroblastoma received the drug biweekly. Twenty-eight patients received an adequate trial, which was defined as a minimum of 5 weeks at the maximal tolerated dose, unless there was progressive disease at the maximal tolerated dose. Side effects of the drug were striking, and included fever, hypotension, myalgia, bone pain, arthralgia, arthritis, abdominal pain, liver toxicity, thrombocytopenia, and neurotoxicity. No complete remissions occurred in spite of interferon levels above 100 U in nearly 50% of patients.

Topics: Acute Disease; Carboxymethylcellulose Sodium; Child; Drug Evaluation; Female; Humans; Interferon Inducers; Interferons; Leukemia; Leukemia, Lymphoid; Male; Methylcellulose; Neuroblastoma; Poly I-C; Polylysine

A comparison was made between the agar and methylcellulose culture systems with respect to their ability to support the clonal growth of leukemic cells obtained from patients with acute myeloblastic leukemia, acute lymphoblastic leukemia, and chronic myelogenous leukemia in blastic crisis. The number of clusters and/or colonies formed and the morphology of the cells within them varied from patient to patient. Nevertheless, no significant difference between the two culture systems within given leukemic specimens was found. No significant differences were noted among three different conditioned media used as sources of colony-stimulating activity. Most of the cells within clusters and colonies were identified to be immature members of granulocyte-macrophage series or to be indistinguishable from the preculture leukemic blast cells by morphological and cell surface marker studies. Cells from myeloid crisis in chronic myelogenous leukemia grew well in the cultures, but cells from lymphoid crisis did not proliferate.

Topics: Adult; Agar; Aged; Antigens, Surface; Cell Division; Cells, Cultured; Female; Growth Substances; Humans; Leukemia; Leukemia, Myeloid, Acute; Male; Methylcellulose; Middle Aged; Rosette Formation

Topics: Animals; Estradiol; Friends; Leukemia; Leukemia Virus, Murine; Leukemia, Experimental; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Mercaptopurine; Methylcellulose; Mice; Mitomycin; Mitomycins; Neoplasm Transplantation; Neoplasms; Pharmacology; Research; Sarcoma; Triethylenemelamine; Urethane