Jouji Hirata

Late‐appearing Philadelphia chromosome in a patient with acute nonlymphocytic leukaemia derived from myelodysplastic syndrome: detection of P210‐ and P190‐type bcr/abl fusion gene transcripts at the leukaemic stage

Summary. We describe a patient with acute nonlymphocytic leukaemia (ANLL) derived from myelodysplastic syndrome in whom the Philadelphia chromosome (Ph1) first emerged at the late stage of ANLL transformation. Cytogenetically, the Ph1 chromosome was not detected until the late stage of ANLL transformation, 14 months after the transformation following a 3‐month history of refractory anaemia with excess of blasts. The cells with and without the Ph1 chromosome had a common abnormal chromosome, t(3;3) (q21;q26). The reverse transcription‐polymerase chain reaction analysis showed no bcr/abl message at diagnosis. However, the mRNA encoding P210bcr/abl was detected in the early stage of ANLL transformation. Furthermore, the mRNAs encoding both P210bcr/abl and P190bcr/abl were detected in the late stage of ANLL transformation when the Ph1 chromosome was detected by cytogenetic analysis. These evidences support a multistep pathogenesis of leukaemias, and the product of bcr/abl fusion gene may influence the course of disease.

Effect of Platelet-Derived Growth Factor and Bone Marrow-Conditioned Medium on the Proliferation of Human Bone Marrow-Derived Fibroblastoid Colony-Forming Cells

The effects of various human sera, platelet lysates and platelet-derived growth factor (PDGF) on the proliferation of human bone marrow-derived fibroblastoid colony-forming cells (CFU-F) were examined. We obtained nearly identical growth curves of fibroblastoid colonies with sera, platelet lysates and PDGF as stimulants and concluded that PDGF was a main growth factor for CFU-F in human serum. In contrast to colony size, CFU-F number was irrelevant to the concentration of PDGF. Removal of culture medium containing hemopoietic cells after short-term incubation of bone marrow cells reduced both colony number and size in CFU-F cultures. When each of bone marrow-conditioned medium (BMCM), peripheral blood mononuclear cells (MNC) and phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM) was added to the cultures, CFU-F number and colony size recovered. The role of PDGF and the factors present in BMCM, MNC and PHA-LCM in the growth of CFU-F and their precursor cells were discussed.

Polyneuropathy in acute megakaryoblastic leukemia

Peripheral neuropathy is a rare complication in leukemia. The authors report a patient with acute megakaryoblastic leukemia (AMKL) and progressive symmetric polyneuropathy. Intense infiltration of leukemic cells in a peripheral nerve was observed at autopsy. This is the first report of AMKL with peripheral nerve involvement to the knowledge of the authors.

Clinical significance of human bone marrow stromal cell colonies in acute leukemias.

Human bone marrow-derived fibroblastoid colony-forming cells (CFU-F) and adipocyte colonies in 36 patients with acute leukemia were studied, and were serially analysed at different clinical stages. At untreated stage, CFU-F number in acute lymphoblastic leukemia (ALL) was lower than that in acute non-lymphoblastic leukemia (ANL). In ANLs, CFU-F number in M1 was lower than that in M2. Adipocyte colonies were frequently developed at regenerating and relapsing stages, but rarely at untreated and remission stages. The adipocyte colony formation did not correlate with any of CFU-F number, marrow cellularity nor number of leukemic cells, but might be associated with hemopoietic regeneration. The favorable prognosis was associated with normal CFU-F number and with adipocyte colony formation at regenerating or bottom stage. As adipocytes in marrow samples were completely removed before cultures, adipocyte colony was probably originated from preadipocytes. Thus, our results suggest that adipocyte precursor cells increase in regenerating marrow and that they are essential in active hemopoiesis.

Deletion of chromosome 6q in two cases of acute myeloblastic leukemia and a review of the literature.

Two cases of acute myeloblastic leukemia (AML M2) associated with a deletion of chromosome 6q are described. One was a 38-year-old man with constitutional inversion of chromosome 9, and another was a 57-year-old female atomic-bomb survivor. The karyotype of these patients were 46,XY,del(6)(q12q14),inv(9)(p11q13), and 47,XX,6q-,+min, respectively. In both cases c-myb protooncogene, which is located in chromosome 6q, was neither deleted nor rearranged, and c-myb messenger RNA level was not elevated. These results suggest that c-myb is not involved in the leukemogenesis of AML with 6q- as well as lymphoid malignancies with 6q-. Out of 23 AML cases with 6q- reviewed, 6 cases had erythroleukemia, and 4 developed in Down syndrome patients.

Pancreatic carcinoma associated with marked eosinophilia: A case report

A case of pancreatic carcinoma associated with marked eosinophilia is reported. A 71‐yr‐old man was admitted to hospital because of melena and abdominal pain. The systematic examinations revealed pancreatic adenocarcinoma with multiple metastases (rectum, lung and brain). The leukocyte count was gradually increased and reached up to 81.7 times 109/1, of which 54% consisted of eosinophils. Colony‐stimulating factor (CSF) was detected both in the patients serum and in the tumor extracts by a normal human bone marrow culture system. The colonies which were stimulated with patients serum largely consisted of granulocyte, granulocyte/macrophage and eosinophil types. These results suggest that blood leukocytosis and eosinophilia were due to a high concentration of plasma CSF, which was probably produced by the tumor cells.

Serial Studies of Bone Marrow-Derived Fibroblastoid Colony-Forming Cells and Granulocyte/Macrophage Precursor Cells in Patients with Acute Leukemia

Bone marrow-derived fibroblastoid colony-forming cells (CFU-F) and granulocyte/macrophage precursor cells (CFU-GM) were studied in patients with acute leukemia. The numbers of CFU-F and CFU-GM were significantly lower in patients with acute myelogenous leukemia (AML) and acute lymphocytic leukemia (ALL) at diagnosis than in normal subjects, although patients with AML had a very wide range of CFU-F colony-forming efficiency. However, a suppressive effect of leukemic cells on normal CFU-F colony formation was not observed. CFU-F and CFU-GM in patients with acute leukemia recovered to normal levels when complete remission (CR) was achieved and decreased again at relapse. Serial studies showed that the increase in CFU-F preceded the recovery of CFU-GM. In AML, furthermore, patients who achieved CR had a higher number of CFU-F than patients without CR, suggesting that the CFU-F level at diagnosis may contribute to the prediction of the likelihood of remission induction in patients with AML.

Periodical Appearance of Erythropoietin-Independent Erythropoiesis in Chronic Myelogenous Leukemia with Cyclic Oscillation1

A patient with Ph1-positive chronic myelogenous leukemia (chronic phase) had a cyclic oscillation in white blood cells, platelets and percent saturation of transferrin. The cycle comprised about 70 days. The number of circulating granulocyte-macrophage colony-forming units (CFU-GM) oscillated with the same phase, while that of bone marrow CFU-GM and erythroid colony-forming units (CFU-E) oscillated in a reverse phase. At the nadir, we observed an abnormal increase in bone marrow endogenous CFU-E (e-CFU-E). An erythropoietin (Epo) dose-response curve of CFU-E showed a high Epo-sensitivity. Anti-Epo rabbit serum did not inhibit the e-CFU-E colony formation. This indicates that Epo-independent erythropoiesis occurs periodically at the nadir. It is suggested that the interactions between the abnormal stem cell and the hematopoietic regulating system cause cyclic oscillation.

Analysis of clonality at the level of progenitors in chronic myelogenous leukemia using the polymerase chain reaction

The Philadelphia (Ph1) chromosome is a specific structural abnormality in which the abl oncogene is activated due to the formation of the novel chimeric gene, bcr/abl. To investigate the clinicopathological role of bcr/abl in Ph1-positive chronic myelogenous leukaemia (CML), we studied the clonal origin of haematopoietic progenitors by detecting bcr/abl mRNA in a single haematopoietic colony using the polymerase chain reaction (PCR). Nine patients with CML were examined. In 5 chronic phase patients, all granulocyte/macrophage (CFU-GM) and erythroid (BFU-E) progenitor-derived colonies were positive for bcr/abl mRNA. Colonies in which the transcripts were not detectable were observed in 4 patients. These 4 patients included one patient with a normal karyotype and without splenomegaly, a patient with cyclic oscillation of her white blood cell level, a patient treated with busulfan and interferon-alpha (INF-alpha), and a patient relapsing after allogenic bone marrow transplantation (BMT). Our observations indicate that detection of Ph1-positive clones by PCR may be used to evaluate clinical stages and the effects of treatment in CML.

Double t(1;7)(p36;p11) in a megakaryocytic crisis of chronic myelogenous leukemia with variant t(5;9;22)

A 56-year-old man with chronic myelogenous leukemia (CML) who presented with a variant Ph chromosome, t(5;9;22)(q13;q34;q11), developed a unique additional chromosomal change of a double reciprocal t(1;7)(p36;p11) during the accelerated phase. A minor clone that had two copies of 1p+ and a copy of 7p- with a normal chromosome 7 was observed simultaneously. The patient underwent a megakaryocytic crisis. Surface marker of the blasts was positive for CD13, CD33, HLA-DR, CD41a, and CD42b, and negative for CD14 and lymphoid markers. Sequential chromosome analysis suggests that the double t(1;7) was caused by a multistep event consisting of duplication of both derivative chromosomes accompanied by loss of normal chromosomes 1 and 7. This may be the first report of a double reciprocal chromosomal translocation in a hematopoietic neoplasm.