Buffy coat autografts for patients with chronic granulocytic leukaemia in transformation

We have treated 20 patients with chronic granulocytic leukaemia (CGL) in transformation with cytotoxic drugs or with cytotoxic drugs and whole-body irradiation followed by transfusion of autologous blood cells collected at diagnosis and stored in liquid nitrogen. The mean number of nucleated cells autografted was 25.1 X 10(8)/kg (range: 12.5-40.1). Full myeloid engraftment occurred in 18 patients; it was partial in one patient and unassessable in another. The median survival for the 20 patients post-graft was 14 weeks. Two patients are alive, one now in recurrent transformation, and one in second chronic phase that has lasted 52 weeks. For the 18 patients who died the mean survival was 24 weeks (range: 2-125). Two patients with predominantly myelosclerotic transformation showed evidence of engraftment. One patient successfully autografted developed features consistent with graft-versus-host disease which proved fatal. We conclude that autografting may offer substantial palliation for some but not all patients with CGL in transformation.     

Cryopreserved peripheral blood cells functioning as autografts in patients with chronic granulocytic leukaemia in transformation.

Six patients with chronic granulocytic leukaemia (CGL) in transformation were treated with cytotoxic drugs or cytotoxic drugs plus total body irradiation, followed by infusion of reconstituted autologous peripheral blood cells that had been collected from them at diagnosis and stored in liquid nitrogen for up to 58 months. In four cases the blood and bone-marrow appearances were rapidly restored to those of typical chronic-phase disease. In three of these patients transformation recurred at 74, 32, and 26 weeks respectively. One patient was still in second chronic phase at eight weeks. One of the patients who entered a second transformation was restored to a third chronic phase by further treatment with cytotoxic drugs and a second autograft. Cryopreserved autologous blood cells may thus restore some patients with CGL in transformation to chronic-phase disease and so may help to prolong life.     

Lymphoblastic lymphoma in a female with chronic granulocytic leukaemia

In a 18 years old, Ph1 negative woman a lymphoblastic lymphoma developed after 16 months of treatment for juvenile form of CGL. In the course of the disease we observed twice the myeloblastic transformation and once the low differentiated transformation of CGL. Based on available methods we are not in a position to exclude a particular form of lymphoblastic transformation of CGL involving lymph nodes without leukaemic blood picture.     

Molecular biology of Philadelphia chromosome in chronic granulocytic leukaemia and acute lymphoblastic leukaemia

In classical t(9;22) translocation, as observed in chronic granulocytic leukemia (CGL), a hybrid DNA unit is produced, including a rearranged PHL gene, previously known as bcr (breakpoint cluster region) plus the translocated c-abl gene from chromosome 9: a hybrid bcr-abl protein, p210 is formed, with increased tyrosine kinase activity. Such DNA rearrangement, with a p210 protein synthesis, is also found in cases of Philadelphia-positive acute lymphoblastic leukemia (ALL), but in apparently similar cases the bcr gene is not rearranged, and a novel p190 abl-related protein can be found; c-abl rearrangement has also been observed.It is thus established that correlations between cytogenetic and molecular events can be found in CGL and ALL, as in other haemopoietic malignancies: translocation and possible rearrangement of the c-abl oncogene seem of particular importance in this case.     

Structural rearrangements associated with the Ph1 chromosome in chronic granulocytic leukaemia

Summary Four cases of C.G.L. in which banding of the Ph¹ chromosome was performed were found to have variation from the usual 9/22 translocation pattern. All 4 cases showed a rearrangement involving at least 3 chromosomes, 2 of which were a 9 and a 22. One of these cases had in addition an XYY karyotype in the bone marrow.     

Cytochemical studies in the blastic transformation of chronic granulocytic leukaemia

The cytochemical features of blast cells were studied in 45 patients with blastic phase of chronic granulocytic leukaemia. Various degrees of Sudan black B positivity was characteristic of myeloblastic transformation (23 patients), while in the medullary blast cells of nine patients with myelomonocytic transformation the alpha-naphthyl-acetate esterase showed intensive activity. In two cases the demonstrability of beta-thromboglobulin and factor VIII-related antigen in blast cells showing otherwise PAS, acid phosphatase and alpha-naphthyl-acetate esterase activity referred to megakaryocytic transformation. In six patients with lymphoid blast crisis proliferation of the Sudan negative blast cells with different granular PAS, acid phosphatase and/or beta-glucuronidase positivity was demonstrated. In five cases the cytochemical findings of leukaemic cells indicated biphenotypic and mixed transformation, respectively.     

The ultrastructural localization of human neutrophil alkaline phosphatase in normal individuals during pregnancy and in patients with chronic granulocytic leukaemia

Summary The intracellular localization of alkaline phosphatase was determined in human neutrophils by electron microscope cytochemistry. In normal individuals, the largest and most intense deposits of reaction product were seen in a unique cytoplasmic granule population termed phosphasomes. Lighter deposits were seen on nuclear membranes, some intracytoplasmic membranes lining vacuoles and granules and occasionally in focal patches on the internal surface of the plasma membrane.In cell isolated from women in the third trimester of pregnancy, activity was found in the same intracellular sites but there were, on average, more alkaline phosphatase-containing granules per cell than in the cells from non-pregnant individuals. Neutrophils from pregnant women were also characterized by the presence of large deposits of reaction product on the external surface of the plasma membrane (extramembranous). This activity had properties characteristic of the placental isoenzyme of alkaline phosphatase, found in serum during pregnancy.Neutrophils from patients with chronic granulocytic leukaemia, showing normal mature morphology, contained significant amounts of granule reaction product but there were fewer phosphasomes per cell than in normal individuals. In morphologically immature cells, reaction product was present in nuclear membrane, endoplasmic reticulum and large granules. These results were in agreement with previous biochemical data confirming a quantitative lack of alkaline phosphatase in chronic granulocytic leukaemia.     

Quantitative cytochemistry of blood neutrophils in myelodysplastic syndromes and chronic granulocytic leukaemia

Quantitative cytochemistry of components of blood neutrophil azurophilic granules (myeloperoxidase, chloroacetate esterase, beta-glucuronidase, and acid phosphatase) and specific granules (lactoferrin) has been performed by scanning and integrating microdensitometry in 13 patients with a myelodysplastic syndrome and 11 patients with chronic granulocytic leukaemia. Both patient groups showed a reduction of enzyme activity in azurophilic granules, and also of lactoferrin, consistent with abnormal development of neutrophil granules. These cytochemical changes in blood neutrophils are similar to those found in acute myeloid leukaemia, are consistent with a leukaemic maturation defect, and may be of diagnostic value.     

Elevation of serum thioredoxin levels in patients with type 2 diabetes.

To evaluate the clinical significance of thioredoxin in diabetic patients, serum thioredoxin levels measured with a recently established sandwich enzyme-linked immunosorbent assay kit were compared with clinical laboratory data and complications in 174 patients with Type 2 diabetes. Thioredoxin levels were significantly higher in diabetic patients (mean value, 38 ng/ml) than in healthy controls (21 ng/ml) (p < 0.05). Fasting blood sugar and hemoglobin A1c did not correlate with thioredoxin. Plasma non-esterified fatty acids levels were significantly higher in patients with higher thioredoxin levels (>or= 40 ng/ml) than in those with lower thioredoxin levels (< 40 ng/ml) (p < 0.001). There was a significant correlation both between thioredoxin and non-esterified fatty acids in patients with diet/exercise therapy (p < 0.01) and between thioredoxin and fasting immunoreactive insulin in those treated with diet/exercise or oral hypoglycemic agents (p < 0.05). Thioredoxin did not correlate with diabetic complications. In conclusion, serum thioredoxin levels may reflect the status of insulin resistance in Type 2 diabetic patients.     

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.     

Karyotypes of 33 patients with clonal aberrations in chronic lymphocytic leukaemia. Review of 216 abnormal karyotypes in chronic lymphocytic leukaemia

We report on 33 unpublished patients with clonal anomalies in chronic lymphocytic leukaemia. The literature was thoroughly reviewed in order 1) to quantify the frequency of anomalies found in chronic lymphocytic leukaemia and to give new status to the rarest, 2) to determine whether a given anomaly was an additional anomaly and/or a primary anomaly, and 3) to find out whether strong associations between different anomalies exist in this disease.