Blast Cells In Peripheral Blood Research Articles

In Vitro and In Vivo Effects of Deferoxamine in Neonatal Acute Leukemia

A six week old infant with acute leukemia failed to attain remission with chemotherapy. Because we previously demonstrated that the iron chelator deferoxamine (DFO) has antiproliferative properties and modulatory effects on cell differentiation, a protocol was designed for in vitro study and for clinical use in the patient. At diagnosis, blast cells were morphologically undifferentiated, had nondiagnostic cytochemistry, showed an abnormal karyotype (t[4;11]), expressed markers of B cell lineage, and demonstrated C mu gene rearrangement. Tissue culture of marrow or blood cells yielded colonies of leukemic blasts. Increasing concentrations of DFO produced a dose-dependent suppression of patient's blast colony growth in vitro, and blasts within colonies showed a marked change in surface antigen expression from lymphoid to myelomonocytic markers, became monocytic in appearance, and developed intense staining for nonspecific esterase. When DFO was given intravenously to the patient as a single agent for 48 hours, blasts no longer expressed lymphoid antigens and became strongly positive for myelomonocytic markers, identical to the in vitro findings. Intravenous DFO halted rising peripheral blood blast cell numbers and allowed a several-fold increase in normal hematopoietic progenitor colony growth. When combined with low-dose cytosine arabinoside in the treatment protocol, DFO caused striking leukemic cytoreduction. Our findings indicate that DFO has antileukemic properties by virtue of its effects on proliferation and differentiation, and they prompt further experimental and clinical studies with this agent.

Prognostic significance of morphologic and cytogenetic findings for progression in myelodysplastic syndromes.

Within the group of myelodysplastic syndromes (MDS) two subgroups are defined [refractory anemia (RA) and RA with ringed sideroblasts (RA-S)] where the percentages of peripheral blood and bone marrow blast cells, by definition, do not exceed 1% and 5%, respectively [1]. Therefore, the applicability of these parameters would be expected to be of limited value with regard to the prediction of progression to RA with an excess of blasts (RAEB) and/or acute myeloid leukemia (AML). The present investigation was undertaken to evaluate the predictive value of peripheral blood cytopenia and to investigate whether a causal relationship between peripheral blood cytopenia, bone marrow morphology, and cytogenetic abnormalities could be envisaged.

Acute leukemia with mediastinal mass, lymphadenopathy, and monocytic precursor cells

Cell marker analysis with monoclonal antibodies (MoAb) as well as differentiation studies with the chemical inducer TPA were used to identify a population of apparently lymphoid cells as monocytic precursor cells in a patient with acute leukemia. The initial manifestation of the disease with mediastinal mass and lymphadenopathy was followed by the appearance of small lymphocyte-like blast cells in bone marrow (BM) and peripheral blood (PB). Although a lymph node biopsy revealed an infiltration with monoblasts, the leukemic cells in BM and PB were not classifiable. However, when the patient relapsed after chemotherapy with large monoblasts and again with morphologically lymphoid blast cells, the latter could be classified by treatment with TPA. After incubation with the inducer (12-48 hr) the cells became positive with the MoAB VIM-D5, showed a strong reaction with alpha-naphthyl-acetate-esterase and developed macrophage like morphology, as well as phagocytic properties. During the terminal phase of the disease, small, lymphocyte-like blast cells predominated. These cells could be classified by a panel of MoABs. They expressed myeloid determinants (VIM-D5, VIM-2, MY 9, VIM-12, VIM-13) but showed no reactivity with MoABs specific for lymphocytic cells.

Oral idarubicin in adult acute leukemia: a preliminary experience.

Twelve patients affected by different types of acute leukemia received idarubicin (4-demethoxy-daunorubicin) by oral route at the total dosage of 45, 60 or 90 mg/m2 distributed over three consecutive days. Drug response was assessed by the decrease in blast cells in peripheral blood and showed some variations between the different types of leukemia. Acute myelogenous leukemia patients and those with blastic crisis of chronic myeloid leukemia appeared to be the more responsive groups; however, the lower dose schedule could explain the less satisfactory results obtained in lymphatic leukemia patients. Data suggest that idarubicin is absorbed rapidly after oral ingestion, in spite of nausea and vomiting, which appeared 3-4 h later and were easily controlled by antiemetic therapy. The purpose of fractionating the drug dosage over three consecutive days is to prolong in the blood an elevated concentration of the main idarubicin metabolite (13-dihydro-derivative), which presents in experimental models an antileukemic potency similar to the parent compound. This study confirms that idarubicin is effective in acute leukemia also by oral route. This formulation could offer some advantages for subjects who cannot tolerate parenteral chemotherapy and it could be proposed in maintenance leukemia protocols.

Natural course of myelodysplastic syndromes--Helsinki experience.

Between the years 1969 and 1984 162 new patients with myelodysplastic syndromes were diagnosed in the Helsinki University Central Hospital. The median survival was 29 months. The survival was best in the FAB‐groups with < 5 % bone marrow blast cells: median 48 months in the RA‐group and 41 months in the RARS‐group. In the FAB‐groups with > 5 % bone marrow blasts, RAEB and RAEBt, the median survival was 21 and 13 months, respectively. In the CMML‐group the survival was almost as short as that in the RAEBt‐group, median 16 months. These differences in survival between the FAB‐groups were highly significant (p = 0.003). Other variables that predicted short survival were Hb < 10 g/1, platelets < 150X 109/1, and the presence of peripheral blood blast cells.26.5 % of all patients progressed to ANLL. The patients in the FAB‐groups with > 5 % bone marrow blasts cells (RAEB, RAEBt) expired more often in ANLL when compared with patients with < 5 % bone marrow blast cells (RA, RARS) and with patients with CMML (p < 0.001). Other variables that predicted progression to ANLL were the presence of peripheral blood blast cells (p = 0.01) and the age < 65 years (p < 0.01). Besides ANLL, infection and/or bleeding were common causes of death. Two thirds of all deaths were due to these causes.

Reversal of γ-Radiation-Induced Leukemogenesis in Mice by Immunomodulation with Thiabendazole and Dinitrofluorobenzene<xref ref-type="fn" rid="FN1">2</xref>

The effect of thiabendazole (TBZ) and dinitrofluorobenzene (DNFB) on radiation-induced leukemogenesis was investigated in the C57BL/6 mouse model. Administration of TBZ-DNFB during, post, or during and post irradiation successfully blocked leukemogenesis, as indicated by the absence of leukemia blast cells in thymus and peripheral blood, as well as prevented thymic lymphoma. TBZ-DNFB treatment prevented the development of leukemia when studies were terminated both after 7 months of last irradiation (disease fully developed) and after 5 months of last irradiation (disease in the process of development). This TBZ-DNFB treatment also resulted in a significant increase in survival.

Myelofibrosis Developing in Megakaryoblastic Leukemia in a Boy with Down's Syndrome

AbstractA 2 1 /2‐year‐old boy with Down's syndrome developed pancytopenia. After spontaneous recovery, leukocytosis with blast cells in peripheral blood and bone marrow developed, and it was associated with thrombocytopenia and splenomegaly. After transient but complete disappearance of all of these findings, they reappeared and a marked fluctuation with 4 subsequent peaks of leukocytosis was observed. Splenomegaly appeared in association with eukocytosis in early phase and persisted after the third peak.Blast cells were identified as megakaryoblasts and had a karyotype of 47, XY, ‐7, ‐1 2, +21, +der(7)t(7p+;?)(p22;?), +der(12)t(6;12p+)(pll;ll).Bone marrow biopsy at the time of recovery from pancytopenia showed hypercellularity with many megakaryocytes and minimal fibrosis. Subsequent serial bone marrow biopsies showed progressively developing fibrosis leading to the final picture of typical myelofibrosis. A histological examination of spleen and an omental tissue showed marked blastic infiltration associated with megakaryocytic infiltration.This case suggests that megakaryoblastic leukemia associated with myelofibrosis is anadditional type of abnormal myelopoiesis seen in Down's syndrome.

Acute Myelofibrosis

Four new cases of acute myelofibrosis are reported, and 63 cases reported in the literature are reviewed. The typical features of this disease include a rapidly progressive clinical course; nonspecific symptoms such as weight loss, anorexia, fatigue and weakness; the absence of organomegaly; pancytopenia; circulating blast cells; and mild abnormalities in the red blood cell morphology. The bone marrow aspirates are usually "dry." The bone marrow biopsies are essential for the diagnosis and show four consistent features: hypercellularity, reticulin fibrosis, proliferation of blast cells and bizarre, atypical megakaryocytes. In 16 cases, the blast cells in peripheral blood and bone marrow, which are unclassifiable by conventional morphology, could be identified as megakaryoblasts by ultrastructural and immunocytochemical techniques. It is concluded that acute myelofibrosis is a definite clinicopathologic entity, which may be related to acute megakaryoblastic leukemia.

Identification of an anti-human osteogenic sarcoma monoclonal-antibody-defined antigen on mitogen-stimulated peripheral blood mononuclear cells.

An anti-human osteogenic sarcoma monoclonal antibody (mouse IgG2b) termed 791T/36 was found to exert complement-dependent cytotoxicity against phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear (PBMN) cells. This reaction was examined by flow cytofluorimetry using indirect membrane immunofluorescence to detect cell-bound antibody and by measurement of the binding of fluorescein-isothiocyanate-conjugated 791T/36 antibody to cells. The antibody reacted strongly with peripheral blood blast cells induced by PHA, and there was negligible reactivity with resting lymphocytes. Maximum binding was observed after 3 days' culture with PHA, coinciding with maximum DNA synthesis, and this represented of the order of 2 X 10(5) antibody molecules bound per cell. After cell surface radioiodination of PHA-stimulated PBMN cells, detergent lysis and immunoprecipitation of antigen with 791T/36 antibody and Sepharose-protein A, the apparent molecular weight of this antigen was determined to be 72,000 by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. This is identical to that of the 791T/36-defined antigen expressed on various osteogenic sarcoma cell lines [3, 17], and by this criterion the antigen is distinguishable from other cell surface markers of activated human lymphocytes.

Terminal Transferase as a Predictor of Initial Responsiveness to Vincristine and Prednisone in Blastic Chronic Myelogenous Leukemia

We undertook a prospective trial to evaluate terminal deoxynucleotidyl transferase activity as a predictor of responsiveness to vincristine and prednisone in 22 Philadelphia-chromosome-positive patients with blastic chronic myelogenous leukemia. Thirteen patients were transferase positive, and nine negative. None of the nine negative patients responded, whereas eight of the 13 positive (P = 0.004) responded with complete clearing of peripheral blood blast cells and a return of normal marrow cellularity with less than 5 per cent blast cells. Among transferase-positive patients under 50 years of age the response rate was 78 per cent. Blast-cell morphology (i.e., lymphoblastic versus myeloblastic) had no significant correlation with either responsiveness or terminal transferase activity. The results of this study suggest that responsiveness to vincristine and prednisone in blastic chronic myelogenous leukemia is confined to patients whose leukemic cells are transferase positive.

Lymphoblastic Conversion in Chronic Myelogenous Leukemia

A child presented with "acute leukemia" in which the blast cells resembled lymphoblasts and had negative cytochemical staining (PAS, Sudan black, and myeloperoxidase). Remission was induced and typical adult-type chronic myelogenous leukemia (CML) followed. Cytogenetic studies initially and during remission and subsequent "acute leukemia" relapses revealed the presence of the Philadelphia chromosome abnormality. Terminal transferase assay performed on peripheral blood blast cells was markedly elevated and soft agar culture growth parameters were typical of acute lymphohlastic leukemia. T and B cell marker studies revealed no markers. This case report with supportive laboratory studies suggests that a cell line with lymphoid characteristics may predominate during acute leukemic transformation. This type of subclassification of leukemia may be of importance in therapeutic planning.

Patterned relationships between IgM and blast cells in peripheral blood in a child with acute leukemia

1. Serum IgM determinations and blast cell counts were performed at regular frequent intervals in a case of acute lymphoblastic leukemia in a child. 2. An inverse relationship tended to exist between the concentration of IgM in serum and blast cells in blood during the course of relapses and remissions. 3. High dosage immunosuppressive chemotherapy caused a marked reduction in serum IgM. 4. A relationship appears to exist between IgM and blast cells which may support the concept that impaired ability to produce antibody to leukemia antigens may be an intrinsic component of this disease.

Terminal deoxynucleotidyl transferase activities in human blood leukocytes and lymphoblast cell lines: high levels in lymphoblast cell lines and in blast cells of some patients with chronic myelogenous leukemia in acute phase

Terminal deoxynucleotidyl transferase, an enzyme which catalyzes the polymerization of deoxyribonucleoside triphosphates, elongating oligo- or polydeoxynucleotide chains, but without direction from a nucleic acid template, is thought to be specific for thymus gland and thymus- derived cells. We have confirmed the observations that high levels are characteristic of thymus gland with both human and calf tissue and that elevated levels may be found in some cases of acute lymphocytic leukemia. High levels were also found in human lymphoblast cell lines with T-cell characteristics, and insignificant activity was observed in leukocytes of patients with chronic myelogenous leukemia not in acute blast phase of the disease, chronic lymphocytic leukemia, human B- cells, and normal human blood lymphocytes even after stimulation with phytohemagglutinin. However, high levels (approximately 200 nmoles/hr/10(9) cells) equivalent to those in thymus tissue and lymphoblast cell lines with T-cell characteristics were found in the peripheral blood blast cells of four patients with chronic myelogenous leukemia in an acute blast phase of their disease. One hypothesis that may explain the present results is that in chronic myelogenous leukemia in acute blast phase of the disease the proliferative blast response may not always be myeloblasts but in some cases it may be lymphoblasts.

Terminal Deoxynucleotidyl Transferase Activities in Human Blood Leukocytes and Lymphoblast Cell Lines: High Levels in Lymphoblast Cell Lines and in Blast Cells of Some Patients With Chronic Myelogenous Leukemia in Acute Phase

Terminal deoxynucleotidyl transferase, an enzyme which catalyzes the polymerization of deoxyribonucleoside triphosphates, elongating oligo- or polydeoxynucleotide chains, but without direction from a nucleic acid template, is thought to be specific for thymus gland and thymus-derived cells. We have confirmed the observations that high levels are characteristic of thymus gland with both human and calf tissue and that elevated levels may be found in some cases of acute lymphocytic leukemia. High levels were also found in human lymphoblast cell lines with T-cell characteristics, and insignificant activity was observed in leukocytes of patients with chronic myelogenous leukemia not in acute blast phase of the disease, chronic lymphocytic leukemia, human B-cells, and normal human blood lymphocytes even after stimulation with phytohemagglutinin. However, high levels (approximately 200 nmoles/hr/10(9) cells) equivalent to those in thymus tissue and lymphoblast cell lines with T-cell characteristics were found in the peripheral blood blast cells of four patients with chronic myelogenous leukemia in an acute blast phase of their disease. One hypothesis that may explain the present results is that in chronic myelogenous leukemia in acute blast phase of the disease the proliferative blast response may not always be myeloblasts but in some cases it may be lymphoblasts.

T-lymphoblastic leukaemia: a distinct variant of acute leukaemia.

Two cases of acute lymphoblastic leukaemia (A.L.L.) of T-cell type are reported. Clinically they were characterized by very high peripheral blood blast cell counts at presentation, the early development of meningeal leukaemia, and relative resistance to treatment with combination chemotherapy. Leukaemic cells from both patients lacked all the B-cell markers investigated, but 60-65% of their cells formed rosettes with sheep red cells. Cytochemical and surface structure studies helped to define additional features of these cells and confirmed their T-cell nature.It seems that this variant of A.L.L. may not be uncommon and that it can be distinguished on clinical and immunological grounds from the usual type of A.L.L. which runs a less aggressive course and lacks B- or T-cell markers.

The turnover of blast cells in peripheral blood after in vitro 3 H-cytidine labelling and retransfusion in human acute leukaemia.

Abstract. The present study was undertaken to investigate the survival and turnover rate of blast cells in the peripheral blood of patients with acute leukaemia. Results from four cases of acute leukaemia and one blast crisis in chronic myelocytic leukaemia are reported. The method used was to label a random sample of leukaemic blast cells (LBC) representative of the total blood LBC population and to observe their behaviour after retransfusion into the patient. 500 ml blood was incubated with 3H‐cytidine in vitro (resulting in 87.3 to 99.7% labelling of LBC), retransfused and serial blood samples taken for 10–20 days. The change in labelling index of LBC was evaluated from autoradiographs and the number of labelled LBC was calculated. Mean grain counts were also determined.From the initial dilution after retransfusion, it appears that the cells are rapidly distributed in a pool some 2 to 5 times greater than that calculated from the blood volume and blast count. In analogy with similar studies on granulocytes, and lymphocytes, it is assumed that the circulating LBC exchange with a marginal LBC pool.The disappearance of labelled LBC can be approximated by a double exponential function with T1/2 values of 1/2 to 2 h and 2.5 to 6 days. Probable reasons for the rapid component are exchange of cells between intravascular and extravascular pools, removal of cells damaged in the labelling process, and sequestration e.g. in the lungs. The slow component is governed largely by the disappearance of LBC from the blood (random loss in addition to senescence) and their replacement by newly produced unlabelled cells. The blood transit times for LBC of 3.7 to 8.5 days, calculated from the slow component, are long compared with those of normal granulocytes and the corresponding turnover rates of LBC are slower than that of normal granulocytes. The blood transit times calculated from the slow component are minimal values since, as a result of cell metabolism, label i8 lost from the LBC so that some cells fall below the threshold of autoradiographic detectability and the curve does not represent only the disappearance of cells. A comparison of the labelling index and number of labelled LBC curves and their changes during the course of the leukaemia and its treatment gives information on the flux changes of LBC since the slope of labelling index depends upon the inflow and outflow of LBC, while the slope of the number of labelled LBC is dependent only on the outflow of cells from the peripheral blood.

Involvement of the erythroid series in blastic crisis of chronic myeloid leukaemia. Further evidence for the presence of Philadelphia chromosome in erythroblasts.

Involvement of the erythroid series in chronic myeloid leukaemia may be evidenced haematologically by a normochromic anaemia with anisocytosis, the presence of circulating normoblasts, the presence of bizarre normoblasts in the marrow and peripheral blood, and rarely the presence of sideroblasts. The Philadelphia (Ph1) chromosome has been assumed to be present in the erythrocyte as well as the granulocyte series on good but circumstantial evidence and it is generally agreed that the leukaemic process probably affects the basic stem cell of the red cells, granulocytes and platelets.When blastic crisis occurs there is an increase in the percentage of blast cells in marrow and peripheral blood, and anaemia, thrombocytopenia and resistance to therapy accompany the apparently irreversible downward clinical course. Aneuploid cell lines emerge (Hammouda, Quaglino and Hayhoe, 1964). We are here presenting evidence that aneuploid Ph1‐positive cell lines are also found in the erythroid cells in blastic crisis and therefore that when this occurs the essential change may also affect the basic stem cell. In a case of chronic myeloid leukaemia seen recently, a sideroblastic phase was found terminally and it was possible to combine standard cytogenetic methods with Perls's prussian blue stain for iron and demonstrate siderotic granules in the cytoplasm surrounding Ph1‐positive aneuploid chromosome spreads. We thus prove the erythroid nature of these spreads, and incidentally provide more direct evidence than offered hitherto, for the presence of the Ph1 chromosome in the erythroid cells.