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PubMed1999

[Expression of bcl-2 gene in the evolution of chronic myelogenous leukemia to blast crisis and its implication].

OBJECTIVE: To investigate the expression of bcl-2 gene and cell apoptosis and cell cycle in bone marrow of chronic myelogenous leukemia (CML). METHODS: APAAP assay and in situ hybridization were used for the expression of BCL-2 protein and bcl-2 mRNA in fresh bone marrow samples from 60 cases of CML. Flow cytometry was used to assess the extent of apoptosis and cell cycle percentage. RESULTS: The expression of bcl-2 gene had no statistical difference between CMLs at presentation and in chronic phase, but was much lower in blast crisis (P < 0.05). The percentage of bcl-2 mRNA positive cells was consistent with BCL-2 protein expression. In addition, BCL-2 protein was related to the Hb levels, BPC, and immature cells in the peripheral blood and bone marrow. Notably, the extent of apoptosis in accelerated phase/blast crisis was much lower than that in chronic phase(P = 0.028), while the cell cycle had no difference. CONCLUSION: High level of bcl-2 gene expression and low extent of apoptosis in bone marrow cells of CML might partially be the mechanism of poor prognosis of blast crisis, and this provides a new experimental basis for early diagnosis and treatment of CML blast crisis.

Adolescent↗
PubMed1977

Chronic myeloid leukemia: physical and functional similarities to acute leukemia.

Blast crisis, closely resembling acute leukemia, is the usual terminal event in chronic myeloid leukemia. Using physical ("fingerprint") and cultural (colony-forming) methods, we have demonstrated distinctive patterns in the stable phase of chronic myeloid leukemia and in blast crisis. An unusual fingerprint alteration preceding the onset of the terminal phase is noted, and cell culture perturbation is evident at different stages of the disease. Our findings indicate that the application of these methods to the study of hemopoietic disorders is valid, and suggest that the use of such techniques may allow a better understanding of the complex cellular events occurring in the course of chronic myeloid leukemia.

Acute Disease↗
PubMed1991

Development of Francisella tularensis antigen responses measured as T-lymphocyte proliferation and cytokine production (tumor necrosis factor alpha, gamma interferon, and interleukin-2 and -4) during human tularemia.

The lymphocyte immune reactivity of 12 tularemia patients to Francisella tularensis antigens prepared from the bacterial cell envelope was examined during a 14-week follow-up study. Lymphocyte blast transformation responses of peripheral blood mononuclear cells (PBMC) to different protein antigens appeared simultaneously 2 weeks after the first symptoms of tularemia, indicating that none of these antigens had any special role at the early phase of immunization. While the lymphocyte blast transformation responses of total lymphocytes to all bacterial antigens were negative in the week 1 samples, continuously growing F. tularensis-specific T-lymphocyte lines were obtained from PBMC at the same time, indicating that an immune response had already occurred. Later, the T-lymphocyte lines and lymphocyte blast transformation responses were similar. Lymphocyte activation among the PBMC was reflected in an increased number of HLA-DR antigen-expressing, CD4-positive T lymphocytes (CD4+ DR+). The mean secretion of soluble CD8 from F. tularemia antigen-stimulated PBMC increased 2 weeks after tularemia onset, but the mean number of CD8+ DR+ T lymphocytes did not vary during the study period and no correlation could be found between the soluble CD8 and number of CD8+ DR+ T lymphocytes. F. tularemia antigen-induced cytokine production was measured from the PBMC supernatants. High levels of tumor necrosis factor alpha were detected from the first week onwards. The highest levels of interleukin-2 and gamma interferon were recorded during the second and third weeks, respectively, after tularemia onset. Interleukin-4 could not be demonstrated in the lymphocyte supernatants.

Adult↗
PubMed1988

Translocation (3;21) in Philadelphia chromosome-positive chronic myelogenous leukemia prior to the onset of blast crisis.

Two female patients in the chronic phase of CML were found to have, in addition to t(9;22), a new karyotypic abnormality--t(3;21)(q26;q22)--present in bone marrow cells. At diagnosis, this abnormality was observed in a small number of marrow cells in both patients, and as the disease progressed in patient 1, the percentage of cells showing t(3;21) was increased, reaching 100% as the transformation to blast crisis occurred. These observations suggest that t(3;21) may represent a new and rare nonrandom rearrangement which may be identified prior to the onset of blast crisis.

Adolescent↗
PubMed2005

Blast count and cytogenetics correlate and are useful parameters for the evaluation of different phases in chronic myeloid leukemia.

Staging of chronic myeloid leukemia (CML) phases is based on cytomorphological criteria that vary considerably between different staging systems. Thus, staging of CML is heterogeneous and causes problems with respect to the comparison of therapeutical strategies and clinical outcome. We evaluated 59 patients with CML in different stages of the disease. In order to define which cytomorphological parameters correlate with cytogenetics we investigated cytomorphology and cytogenetics in parallel in all cases. As a result, bone marrow blast count demonstrated a highly significant correlation with the respective cytogenetic results of the patients and was clearly linked to the frequency and complexity of clonal evolution. We therefore propose to focus staging systems of CML on the correlation of the percentage of bone marrow blasts and the cytogenetic results.

Blast Crisis↗
PubMed2004

[Allogenic bone marrow transplantation in chronic myeloid leukemias].

AIM: To assess the role of allogenic bone marrow (ABM) transplantation in chronic myeloid leukemia (CML). MATERIAL AND METHODS: 44 ABM transplantations were performed in 37 CML patients in the chronic phase and 7 patients in acceleration or blast crisis. RESULTS: A complete molecular remission was achieved in 26 (59%) patients: 67.6% after ABM transplantation in the chronic phase and only 14.3% after myelotransplantation in non-chronic phase. Follow-up was 8-150 months (median--59 months). Early lethality after ABM transplantation in the chronic phase was under 14%. A phase of the disease plays a key role in ABM transplantation. If it is made in a chronic phase, CML recurrence rate is low (in our series it was 14%), efficacy of donor's bone marrow lymphocyte transfusions is high. The second complete molecular remission was achieved in 3 of 4 cases of posttransplantation recurrences. Probability of maintenance of a complete remission after ABM transplantation in a chronic phase was 75%, recurrence-free survival--64%, uneventful survival 55% for 90 months. CONCLUSION: The experience of many years demonstrates high efficacy of ABM transplantation in the treatment of chronic myeloid leukemia. It promotes long-term molecular remission the maintenance of which did not require therapy in 65% patients.

Adolescent↗
PubMed2003

Solid phase micro extraction coupled with on-column GC/ECD for the post-blast analysis of organic explosives.

Gas chromatographic analysis with electron capture detection is very sensitive to post-blast residues and useful for the determination of organic explosive molecules. But many compounds extracted from the matrices may interfere with the explosives. Using SPME, most interfering compounds are eliminated so the identification is easier. Another advantage of the technique is a low limit of detection. In this study, four different SPME fibers were tested to analyze the most common encountered organic explosives including nitro aromatics, nitramines and nitro-esters. Different parameters were tested (desorption time, agitation, ...) and a special device has been created to optimize the agitation. Direct desorption effect of the SPME fiber on the column compared to normal split-splitless injection is shown. In this way, the degradation of the most sensitive molecules is decreased. An application to a real case is also described in this paper.

Journal Article↗
PubMed1997

[The methylation of CT gene in chronic myeloid leukemia].

To investigate the relationship between the abnormal methylation of the calcitonin (CT) gene and the karyotypic abnormality in predicting the disease progress in chronic myeloid leukemia (CML), we studied the abnormal methylation of the CT gene in 47 CML patients by using a sensitive PCR method. At the same time, cytogenetic study was made in the same group of patients. The abnormal methylation of CT gene was found in only 4 of 24 patients in chronic phase, but in 6 of 9 patients in accelerated phase, and in 12 of 14 patients in blast crisis. Sequential studies in one patient also showed that the gene is normally methylated during the chronic phase but turns hypermethylated as the disease progresses. Our findings indicate that abnormal methylation of the 5' region of the CT gene is regularly a marker of disease progression in CML.

Adolescent↗
PubMed1987

Cytophotometry of granulocytes in chronic granulocytic leukaemia patients. Part II. Effects of chemotherapy on cell kinetics and cytochemical parameters of granulocytes.

A successful course of chemotherapy (myelosan or combined regimens) results in a two to eight fold decrease of the percentage of S + G2 cells in the bone marrow and peripheral blood and in the restoration of normal morphometric and cytochemical patterns of polymorphonuclear neutrophils in the chronic phase of CGL. In those patients of the chronic phase not responding to chemotherapy and in blast crisis the proportion of S + G2 cells did not change after a course of chemotherapy. Cytostatic drugs killed cells in all phases of the cell cycle. The initial response to chemotherapy was a slight increase of the proportion of S + G2 cells in peripheral blood, apparently as a result of the block in cell cycle transition which was followed by a rapid drop of S + G2 proportion. During chemotherapy such phenomena as an appearance of tetraploid mature neutrophils and of aneuploid clones were occasionally observed.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1994

Absence of N-ras mutations in myeloid and lymphoid blast crisis of chronic myeloid leukemia.

Mutations within N-ras oncogene codons 12, 13, and 61 occur in approximately 25-30% of patients with acute nonlymphocytic leukemia and at a lower frequency (6-20%) in patients with acute lymphocytic leukemia. Moreover, N-ras mutations have been described in patients with chronic myeloid leukemia (CML) in blast crisis but have not been observed during the chronic phase of the disease. In view of the morphological and clinical similarities between acute leukemia and the blast crisis of CML, the question was raised whether the presence of N-ras mutations is associated with the phenotype of acute leukemia. We investigated leukemic cells from 100 patients with CML for the presence of N-ras mutations in the mutational hot spot codons. The cases analyzed included 87 diagnosed with different types of blast crisis and 13 cases in accelerated or chronic phase of the disease. Fragments from N-ras exons I and II containing the codons of interest were amplified by polymerase chain reaction and analyzed for the presence of point mutations by three different technical approaches, including specific oligonucleotide hybridization, direct sequencing, and single-strand conformation polymorphism analysis. N-ras mutations were not detected in any of the CML patients investigated. Only one patient, in whom the initial diagnosis of CML-blast crisis had been revised to chronic myelomonocytic leukemia, displayed an N-ras mutation within codon 13. Our data strongly suggest that N-ras mutations do not play a role in myeloid or lymphoid blast crisis of CML.

Base Sequence↗
PubMed1993

Correlation of marrow iron patterns with disease status of chronic myelogenous leukemia.

Greater than 80% of patients in the stable phase of chronic myelogenous leukemia (CML) have no detectable stainable iron in the marrow yet have normal serum iron, total iron binding capacity, and serum ferritin values. We studied the pattern of marrow iron in 187 marrow aspirates from 67 patients with Philadelphia chromosome positive CML in the chronic, accelerated and blast crisis stages. Sequential marrow aspirates in 30 patients confirm that the switch from negative iron to positive iron is coincident with the development of the accelerated phase of blast crisis of CML. Our data suggest that the pattern of marrow iron staining in CML is a useful prognostic indicator of the disease evolution.

Adult↗
PubMed1990

[Importance of DNA-cytometric parameters in the prediction of blast crisis in the course of chronic myelogenous leukemia].

In a retrospective study the DNA-cytometric parameters stemline ploidy, stemline shoulder fraction and proliferative fraction were followed during the course of the disease in 20 patients with chronic myelogenous leukemia. Stemline ploidy and stemline shoulder fraction significantly increased whereas the proliferative fraction steadily decreased during the disease most of these changes taking place during chronic phase before the clinical onset of blast crisis. Prognostically relevant cutpoints indicating disease transformation during the next 12 months could be defined.

Blast Crisis↗
PubMed1984

Autologous marrow transplantation for patients with chronic myelogenous leukemia (CML) in blast crisis.

Even patients with chronic myelogenous leukemia (CML) in blast crisis were treated with chemotherapy, followed by infusion of autologous bone marrow that had been collected during the chronic phase of the disease and cryopreserved at -198 degrees C. The mean age of the nine females and two males in this study was 34 years with an average duration of the chronic phase of the disease of 5.5 years. Seven out of the 11 patients had a splenectomy prior to intensive chemotherapy. The median survival of the first four patients who received 6-thioguanine, cytosine arabinoside, daunorubicin (TAD) chemotherapy was 2.6 weeks and no patient reachieved the chronic phase of CML. The second group of seven patients received more intensive chemotherapy (MAdHAT), which included melphalan 30 mg/m2 days 1, 2, and 3; Adriamycin 50 mg/m2 intravenously (iv) day 1, hydroxyurea 1500 mg/m2 by mouth for 5-7 days, cytosine arabinoside 100 mg/m2 continuous infusion for 5-7 days, and VM-26 100 mg/m2 iv on day 3. Six out of these seven patients reachieved chronic phase CML after bone marrow reinfusion. The median survival was 29.9 weeks for all patients and 33 weeks for the six patients who reachieved chronic phase CML. All patients subsequently died of recurrent blast crisis. There was no correlation between the time of bone marrow storage and the duration of subsequent chronic phase CML. These studies have shown that autologous bone marrow transplantation after high-dose chemotherapy can result in bone marrow engraftment with reestablishment of chronic phase CML, and prolongation of survival.

Adult↗
PubMedDate not supplied

Trypanosoma cruzi: the expansion of NK, T, and NKT cells in the experimental infection.

T and NK cells play a key role in resistance to Trypanosoma cruzi infections, mainly through IFN-gamma production. The expression of T and NK cells surface markers was studied in NWNA spleen cells of resistant C3H and susceptible BALB/c mice that release IFN-gamma in the early and late acute infection, respectively. In the progressively enlarged spleens, we found: (a) an increased percentage and number of NK blast cells as early as at 2 days post-infection (pi), (b) an enrichment of T and NK cells, in both the total and blast populations, during the late acute phase. At 17 days pi, there was also an accumulation of TCR- alphabeta+DX5+, NKT cells, mainly in resistant mice. At 21 days pi, the enrichment of NK cells ceased, while spleen cells and the T cell compartment continued their expansion. In the chronic stage, TCR-alphabeta+ blasts were expanded in both mouse strains, but NK blasts increased only in BALB/c that, unlike C3H mice, release IFN-gamma. As T and NK cell proliferation is not always associated to IFN-gamma release the experimental downregulation of their expansion to avoid tissue damage could be explored.

Animals↗
PubMed1986

Further characterization of the in vitro tumoricidal activity of staphylococcal protein A.

Treatment of plasma or serum from leukemic patients with solid phase staphylococcal Protein A induced leukemic blast cell lysis in vitro, but this effect was relatively independent of the amount of immunoglobulin G (IgG) removed. Samples with approximately equal cytotoxic activity contained markedly different IgG levels, while samples with similar IgG levels had a wide range of tumoricidal activity. Assays of plasma samples collected during a perfusion of one plasma volume through a Protein A-Sepharose column indicated that the duration of the procedure had a greater effect on cytotoxic activity than did the amount of IgG removed. Neither added leukemic nor normal IgG significantly improved blast cell viability in treated serum. Cytotoxic activity was not dialyzable and concentrated in the Mr less than 100,000 fraction of samples separated by filtration. Treated cytotoxic serum samples did not have important Clq binding activity. These results suggest that the in vitro tumoricidal activity of solid phase Protein A is probably due to a toxic substance added to serum during immunoadsorption rather than to its immunoadsorptive capacity.

Antigen-Antibody Complex↗
PubMed1993

Effects of interleukin-11 on the proliferation and cell cycle status of myeloid leukemic cells.

Interleukin-11 (IL-11) is a pleiotropic cytokine with effects on many different targets. Within the hematopoietic system, the effects of IL-11 are largely manifest only through combination with other cytokines, including IL-3 and Steel factor (SF). In the present study, we addressed the question of IL-11 responsiveness within the different types of human leukemic cells, as well as the mechanism of action of IL-11 at the cellular level. Analysis of a panel of samples from different patients with acute myeloblastic leukemia (AML) and myeloid leukemic cell lines indicated that IL-11 alone was ineffective in supporting myeloid leukemic cell growth but frequently enhanced growth supported by IL-3, granulocyte-macrophage colony-stimulating factor (GM-CSF), or SF. In contrast, three acute pre-B lymphocytic leukemia (pre-B-ALL) and two acute T lymphocytic leukemia (T-ALL) lines failed to respond to IL-11 alone or when combined with other cytokines. The growth enhancement of IL-11 among the AML patient samples was dose dependent and remarkably constant with half-efficient concentrations in the range of 0.3 to 0.4 ng/mL. The thymidine suicide studies with the patient samples revealed that 40% to 50% of the blast cells were in S-phase when exposed for 16 hours to IL-3 and this level was increased to 70% to 90% in response to either IL-11 or IL-6. Our data suggest that the latter two interleukins act synergistically with the direct mitogenic factor, IL-3, in triggering AML blast-cell proliferation. Detailed analysis with several patient samples further revealed that SF and IL-11 both enhance IL-3-supported clonogenic growth of AML blasts and the combination of all three growth factors yields optimal growth. In contrast, IL-6 does not further enhance the effect of IL-11. These results indicate that SF and IL-11 enhance IL-3-dependent clonogenic growth through two distinct pathways, whereas IL-6 and IL-11 may trigger the same pathway.

Cell Cycle↗
PubMed1984

Chronic myeloid leukaemia in Singapore: a case study of 56 patients.

A case study was made on 56 patients with chronic myeloid leukaemia in Singapore. The most common clinical presentations were abdominal discomfort with swelling and hepatosplenomegaly. The mean haemoglobin, white cell count, and platelet count at presentation were 9.23 g/dl, 217.9 X 10(9)/1 and 418 X 10(9)/1 respectively. Twenty-three of the 56 patients had gone into blast transformation at the time of analysis. The median duration of the chronic phase from the time of diagnosis to the time of blast transformation was 23 months. The median duration of survival after blast transformation was 3 months. The median duration of survival for all patients was 23 months. Busulphan remains the drug of choice for the chronic phase of chronic myeloid leukaemia.

Adolescent↗