Stromal cell growth and blast colony-forming cells in AML bone marrow.

Bone marrow cells (BMC) from normal donors and from patients with acute myeloid leukaemia (AML) were cultured. Growth kinetics and the efficiency of stromal layers in supporting the adhesion of normal blast-colony forming cells (BL-CFCs) were studied. BMC from treated AML patients formed confluent stromal layers faster than normal BMCs. BL-CFC binding capacity of normal and AML stromal layers did not differ: on normal stromal layers 67.3-147, on AML stromal layers 63-117 colonies per 5 x 10(5) plastic non-adherent BMC were formed. The amount and/or binding capacity of BL-CFCs was found to be normal in two AML patients in complete remission, while a significantly reduced number and/or binding capacity of BL-CFCs was found in AML non-treated patients and in patients within 4 weeks after the last cytostatic course.

Antibodies to primate retrovirus antigens in circulating immune complexes of patients with acute myeloid leukemia.

Circulating immune complexes were isolated from sera of 8 patients with acute myeloid leukemia (AML) in relapse, and 20 healthy blood donors. F(ab')2 fragments were prepared from the isolated complexes. Using a radioimmunoassay (RIA), these F(ab')2 fragments, the undigested complexes and the original sera were examined for the presence of antibodies against a panel of primate retrovirus antigens: gp70, p15 and p30 of gibbon ape leukemia virus (GaLV) and baboon endogenous virus (BaEV). F(ab')2 fragments derived from the immune complexes of all patients reacted with one or more of the antigens tested, whereas no antibody activity was found in the sera or undigested immune complexes of the same patients. By a competitive RIA, antigens related to GaLV and/or BaEV were found in the serum of 7 out of 8 patients. No markers of these retroviruses were detected in the F(ab')2 preparations, in immune complexes or in sera of any of the 20 control subjects. Our results indicate that a part of the circulating immune complexes in AML contain antigens related to primate retroviruses and specific antibodies to these antigens.

Alpha-1-antitrypsin-induced inhibition of complement-dependent phagocytosis.

In a previous investigation, inhibition of complement-dependent rosette formation by alpha 1-antitrypsin (alpha 1-AT) was observed, and it was demonstrated that alpha 1-AT interacts through its carbohydrate portion with C3 and its fragments. In the present study, the effect of alpha 1-AT on the complement-receptor-mediated phagocytosis by human peripheral blood monocytes was examined. Purified alpha 1-AT inhibited in a dose-dependent manner phagocytosis of C3-carrying yeast particles. Inhibition was selective, concerned only C3-receptor-mediated phagocytosis, neither Fc-receptor-mediated phagocytosis nor uptake of untreated yeast particles was blocked by alpha 1-AT. It was demonstrated that alpha 1-AT towards C3 and fragments of C3 was not mediated by its antiprotease effect, but by its carbohydrate moiety. This finding suggests that alpha 1-AT may have an impact on various immune functions involving complement receptors.

New set-up for capillary isoelectric focusing in uncoated capillaries.

Substituted aminomethylphenol dyes, low-molecular-mass isoelectric point (pI) markers and hemoglobin samples from normal individuals and diabetic patients were used to test a new set-up of capillary isoelectric focusing (cIEF) in uncoated capillaries. In previous cIEF methods, a mixture of sample components and carrier ampholytes was applied in the capillary and analyzed. In the new set-up a fractionated injection protocol is used to apply a 'sandwich' ampholyte-sample-ampholyte plug in the capillary for analysis. This new set-up allows the separation of amphoteric compounds having pI values outside the pH region of the ampholytes applied in the capillary with high precision. The high resolution power of this technique was proven with the analysis of hemoglobin variants.

[Granulocyte-macrophage progenitor cells in allogenic bone marrow transplantation: correlation of progenitor cell content and regeneration in the graft]. / Granulocyta-makrophag össejtszint vizsgálata allogén csontvelöátültetésben: az oltvány össejttartalma és a regeneráció összefüggése.

The amount of granuloid macrophage progenitors (CFU-GM) was studied in 16 donor bone marrows used for allogenic bone marrow transplantation in the National Institute of Haematology and Blood Transfusion between January, 1984 and January, 1988. In 10 bone marrow transplanted patients long-term follow up of bone marrow CFU-GM regeneration was carried out. Graft sizes were the following: 2.91 +/- 0.62 X 10(8)/kg body weight nucleated cells and 19.2 +/- 14 X 10(4)/kg body weight (CFU-GM. Preinfusion procedures (centrifugation and resuspension) did not alter CFU-GM content of the grafts. Separation of nucleated cells with hydroxyethylstarch, applied for ABO mismatched donor bone marrow, however, resulted in a 30 per cent loss in CFU-GM. Since higher than threshold graft-sizes for successful engraftment were used, no linear correlation between graft size and speed of granulocyte and platelet recovery was found. Significant difference between regeneration kinetics of bone marrow CFU-GM of patients transplanted for CML or AML and ALL was observed: in AML and ALL patients normal bone marrow CFU-GM level was found 4 to 6 months after transplantation, while in CML patients CFU-GM level approached the lower limit of the normal value only 10 to 14 months after transplantation. Granulocyte and thrombocyte recovery of CML patients showed a significant delay when compared to transplanted AML and ALL patients.

[Cytochemical, immunologic and gene rearrangement studies in adult acute leukemia]. / Cytochemiai, immunológiai és génátrendezödés vizsgálatok felnöttkori akut leukaemiákban.

Results of morphological, cytochemical and immunological studies performed in adult acute leukaemias have been compared. Thirty one cases proved to be acute myeloid leukaemia, while 25 cases were shown to be acute lymphoid leukaemia. Based on our results we conclude that immunophenotyping with monoclonal antibodies does not help in distinguishing the subtypes of AML. For purposes of clinical diagnosis cytochemical methods are valuable. On the other hand the monoclonal antibodies are essential in distinguishing the very immature myeloid and lymphoid leukaemias and this is of great importance from the clinical point of view, in determining therapy. Moreover, the diagnosis of acute lymphoid leukaemias is not possible without the specific monoclonal antibodies. Their application is first of all in haematological centers caring for leukaemia patients nowadays already obligatory. Gene rearrangement studies make the diagnosis more accurate and help in the diagnosis of leukaemias of unknown immunological origin.

[Gene rearrangement in leukemia]. / Génátrendezödés leukaemiákban.

In this report we summarize our experiences based on the gene rearrangement study of 111 leukaemic patients of different kind. The lymphocyte DNA of the patients was studied for rearrangement of the immunoglobulin light chain constant-, the heavy chain joining- and the T cell receptor beta chain constant region. Our data have well supplemented the results of the monoclonal antibody experiments. In 33 cases the DNA study was in good agreement with the immunological data. In 42 our data helped in gave different results, immunological results. In 11 cases evaluating the DNA and immunological data indicating the necessity of further investigation. The results were inconclusive in 25 cases. As a conclusion we consider the gene rearrangement study to be useful for diagnostic purposes.

[Importance of the detection of minimal residual disease in the management of acute leukemia]. / Minimális reziduális leukaemia vizsgálatának jelentösége az akut leukaemiás betegek kezelésében.

Between 1984-1988, 57 adult acute leukemic patients were treated with intensive combined chemotherapy in the National Institute of Haematology and Blood Transfusion. For the evaluation of response to therapy, 4 investigations were performed in parallel: bone marrow aspirate, bone marrow biopsy, cytogenetic analysis and bone marrow culture. Nonparametric test for samples taken for the evaluation of remission status showed that bone marrow biopsy was significantly the most sensitive method for the detection of residual disease. The bone marrow culture was also on the borderline of significance, but the low CFU-GM level did not always correlate with the further clinical course. Occasionally, karyotypic abnormality was the only sign of the residual disease. It would be of great importance to quantitate the minimal residual disease in order to evaluate and compare the various intensive postinduction therapeutic strategies.

[Experience with the molecular genetical detection of the chimera gene of the Philadelphia chromosome]. / Philadelphia chromosoma chimera génje molekuláris genetikai kimutatásának tapasztalatai.

Molecular genetical techniques could be developed for detection of the chimera gene of Philadelphia chromosome or that of its gene product, due to the relatively conserved structure of the chimera gene. The authors successfully analysed 123 blood/bone marrow samples from 106 patients using these molecular techniques adapted from the literature. Patients were classified by the first diagnosis, 65 CML, 7 AML, 13 ALL patients were studied. 12 patients had the diagnosis of myeloproliferative syndrome, and 9 patients were after bone marrow transplantation. 57% of the total, and by diagnosis, 74% of CML, 28% of AML, 54% of ALL, and 33% of post-transplant samples have shown the chimera gene structure characteristic for Philadelphia chromosome. All patients of myeloproliferative syndrome were negative. In some cases the authors had the opportunity to study simultaneously the peripheral blood and the bone marrow sample of the same patient and of the same date. The ratio of the positivity of the two samples varied from one to infinite. The authors could follow the effect of interferon in one case, the change of clonality of the leukemic cell line in an other case. They had the opportunity to detect two different abnormal gene structures in the sample of an AML patient.

Cell surface markers in chronic lymphotic leukaemia.

Surface markers were applied for the study of the peripheral lymphocyte population in chronic lymphotic leukaemia. The proportion of B-lymphocytes recognized by EAC-rosetting and by surface-bound Ig was found higher in chronic lymphotic leukaemia than in normal controls. On the other hand, the E-rosette test revealed a very low proportion of T-cells. No relationships were demonstrable between the surface markers and the clinical features.

Combination chemotherapy in adults with acute leukaemia.

Ten patients with ALL and 35 with AML received combination chemotherapy at out Institute during the period 1972 - 1975. Patients with ALL were treated according to the L-2 protocol, those with AML according to the L-2, MRC 5/A, MRC 5/B and TRAP protocols. Nine of the 10 patients with ALL entered complete remission, and median survival time of these patients was 9.2 months. Four of the 35 patients with AML achieved complete, 13 partial remission. Median survival in this group was 4 months. Infection and bleeding represented the most frequent and severe forms of complication. Bleeding was well controlled by the administration of platelet concentrates, but infections were often lethal. The poor results were probably due to the high incidence of severe infections.

The treatment of infections in granulocytopenic patients with granulocyte transfusions.

Twenty-three severely neutropenic patients were given 31 courses of ABO and HL-A compatible granulocyte-rich plasma transfusions between 1974--76 at the National Institute of Haematology and Blood Transfusion. The criteria for the effectiveness of therapy were disappearance of fever and survival for over 3 weeks after granulocyte transfusion. Of the 28 evaluated cases granulocyte transfusions were effective in 16 and ineffective in 12 cases. The effectiveness of granulocyte transfusion was favourably influenced by early introduction of therapy, and continuation for several days.

Studies on the composition and elimination of circulating immune complexes in leukaemia.

Longitudinal studies for the detection of circulating immune complexes (CIC-s) were performed in leukaemic patients using three methods in parallel. Results of these studies indicated that immune complexes in leukaemia are heterogeneous and their composition changes in the course of the disease. These results were supported by the changes in the concentration of a complement component (C1) and in the IgG subclasses in the immune complex-enriched fraction prepared by polyethylene glycol precipitation from different serum samples of individual patients with leukaemia. CIC-s from leukaemic patients are normally engulfed by granulocytes of healthy donors. The phagocytic capacity of leukaemic blast cells is hampered. The phagocytic activity through Fc and C3 receptors of peripheral mononuclear cells of leukaemic patients did not run parallel. On the basis of these studies it may be surmised that the total phagocytic capacity of the leukaemic patients cannot keep pace with the increased rate of immune complex formation.

Ganglioside composition in common acute lymphoblastic leukaemia.

Human leukaemic lymphocytes of the common acute lymphoblastic leukaemia (cALL) subtype were analysed for ganglioside composition by over pressured layer chromatography-densitometry. Two major ganglioside components, GM3 (58.0-77.8% of the total sialic acid content) and GM1 (20.7%-29.2%), and some minor compounds, GM2 (trace amounts to 9.6%) and unidentified gangliosides with chromatographic mobility between GM1 and GD1a (trace to 12.6%), were isolated from these cells. The relative amount of GM3 in cALL cells was found to be about double that in normal lymphocytes (65.7% vs. 37.7%), but lower than in chronic lymphocytic leukaemia (CLL) cells (65.7% vs. 81.5%). The cALL cells contained trace amounts of the ganglioside GD3, a compound present in CLL cells (5.6%) but absent in normal lymphocytes.