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J Soucek

Publications and source records attributed to J Soucek.

At least 37 records · Page 2Linked to original sources
PubMed1992

[Results of bone marrow transplantation at the Institute of Hematology and Blood Transfusion in Prague].

The first allogenic bone marrow transplantation (TKD), when for the preparation whole body irradiation was used, was implemented in the Institute of Haematology and Blood Transfusion (UHKT) in Prague in 1986. Before June 1992 36 TKD were performed incl. 28 allogenic, 2 syngenic and 6 autologous. For the first time bone marrow from a non-related donor was transplanted. Of 30 allogenic and syngenic TKD to the present time 17 patients survive, i.e. 56.6% of the whole group. According to individual diagnoses 8 patients with the diagnosis of chronic myeloid leukaemia (CML) survive, 5 of 10 patients with the diagnosis of acute leukaemia (AL) and 3 of 4 patients with the diagnosis of severe aplastic anaemia (SAA) or with Fancon's anaemia (FA) resp. The survival period of the whole group is from 1-62 months since the transplantation. The main cause of death of 8 from 13 patients who died were infections associated with acute or chronic disease of the graft against the host (GVHD). In autologous TKD the bone marrow was treated with etoposide. Of the six transplanted patients with AL five survive 1.5-30 months after transplantation. The authors present some general information of pretransplantation preparation, prevention of GVHD, its incidence and results of TKD.

Adolescent↗
PubMed1991

Induction of NK and LAK activities in human lymphocyte culture by a cytosol fraction from leukemic myeloblasts and by monoclonal antibody CD 3.

The stimulating effect of cytosol fraction (F3) isolated from human myeloblasts (m.w. ranging from 30 to 100 kDa) and monoclonal antibody CD 3 (MEM 57) was tested on NK and LAK cell activities in peripheral mononuclear cells (PMNC) of normal donors and leukemic patients. The F3 fraction added in 10% of the total volume of RPMI 1640 medium induced significantly increased lytic activity of normal lymphocytes or IL-2 activated lymphocytes against K 562 cells in 3-day culture. Similarly, the proliferation of both cell cultures was enhanced by F3. The effect of F3 on NK and LAK cell activities in culture of PMNC from leukemic patients was less pronounced and synergic action of F3 did not occur, whereas the proliferation of leukemic cells was significantly enhanced. MEM 57 increased the cytotoxicity and cell proliferation in 3-day culture of normal lymphocytes at concentrations ranging from 10 to 250 ng/ml. MEM 57 stimulated also NK cytotoxicity estimated in freshly isolated normal lymphocytes. The deficient NK cell activity observed in PMNC of leukemic patients was induced by MEM 57 or its combination with IL-2 in 3-day culture. These observations indicate that combination of more immunomodulating agents can lead to shortening of the incubation time necessary for correction of the NK cell defective activity in PMNC of leukemic patients.

Adjuvants, Immunologic↗
PubMed1990

[Changes in NK cell activity in patients with acute leukemia].

Examination of the activity of NK-cells in peripheral mononuclear cells (PMNB) of patients with acute leukaemia (AL) was implemented before the onset of treatment and in the course of the disease following cytostatic treatment. The examination was made by the isotope method based on the release of 51Cr after 4-hour cultivation of effector cells with target cells of line K 562. The mean percentage of NK cellular activity in patients with AL was 9.9 and 5.9, as compared with normal values of 56 and 41 with ratios of effector and target cells of 40:1 and 20:1. The course of the disease after cytostatic treatment revealed that patients where complete remission occurred had before onset of treatment on average a higher NK-cell activity than those who responded poorly to treatment. In the majority of patients who were in complete remission the activity of NK-cells increased markedly.

Acute Disease↗
PubMed1990

[Immunologic monitoring of bone marrow transplantation. Experience with 16 cases].

HLA-A, B antigens and the ABO group were examined in 184 patients with aplasia of bone marrow and leukaemia and in 373 of their relatives, mostly siblings. A HLA-A, B donor, identical or compatible, was found for 35.87% patients, a HLA-DR identical for 84.21% of 38 patients who had a HLA-A, B identical relative. Bone marrow was transplanted to 16 patients (10 with bone marrow aplasia, 6 with acute or chronic leukaemia), with one exception bone marrow from a sibling. The bone marrow was accepted in all patients but two where the transplantation was made despite MLC positivity. From the results ensues that it is essential for successful transplantation of bone marrow to ensure maximal identity between donor and recipient as regards the ABO group, HLA antigens and negativity of the MLC reaction. The negativity of the MLC reaction is more important than HLA-DR identity; when assessing only one HLA-DR antigen in a donor identical with the patient, it cannot be ruled out that on the lymphocytes of the donor there exists another one which was not detected. The authors discuss the causes of different results of the MLC reaction and HLA-A, B, DR typing.

ABO Blood-Group System↗
PubMed1989

[A simple model for verifying the effects of a focused shock wave from our generator on erythrocytes and lymphocytes].

Biological effects of the focused shock wave produced by the generator of our own construction were observed on the erythrocyte and lymphocyte suspension as a model. Erythrocyte haemolysis located into the small volume of the 1.4 ml focus was observed in the dependence on the number of shocks. After 10 shocks it does not exceed 15%. In the previous experiments this particular dosage caused a complete destruction of the bile stone of about 1 cm size. Increase of kalium concentration in correspondence to the degree of haemolysis was discovered in the erythrocyte suspension. Effects of shock wave were observed also in lymphocytes, where no change of blastic transformation in the decrease of the life span approximately by 2% was proved. Regular measurements proved minimum damage of blood cells in given experimental conditions and the results are comparable with biological effects of other sources of the focused shock wave.

Erythrocytes↗
PubMed1989

Positive effect of direct current on cytotoxicity of human lymphocytes.

Normal or leukemic human lymphocytes treated with direct current (DC) showed enhanced antileukemic cytotoxicity. The enhancing effect of DC-treated lymphocytes was dependent on current density and time exposure. A desirable effect was achieved with current densities ranging from 5 to 10 mA/cm2 at a short exposition time (5--10s). Enhanced lymphocyte cytotoxicity occurred after a 48 h cultivation at 37 degrees C in a humidified atmosphere containing 5% CO2 and was proved by the increased number of trypan blue stained target cells, tumor-binding cells, and lymphocytes with activated nucleoli. Lymphocyte cytotoxicity measured immediately after DC-treatment was not enhanced. Furthermore, the cytotoxic effect was potentiated using media conditioned with interleukin-2 (IL-2) or cytosol fraction (F3) isolated from human leukemic cells. Such in vitro stimulated cytotoxic cells displayed reactivity against K 562 cells as well as fresh leukemic cells of allogeneic origin. Of considerable clinical interest is the observation that lymphocytes treated with DC in IL-2 or F3 conditioned media may enhance antileukemic cytotoxicity in peripheral lymphocytes of patients with hematological malignancies.

Cytosol↗
PubMed1987

Features of immaturity in cells derived from granulocytic differentiation inducer treated human myeloid leukaemia (ML-1) cells.

Cells of the human myeloid leukaemia cell line ML-1 were exposed to differentiation inducing doses of dimethylsulfoxide (DMSO) and retinoic acid (RA). DMSO (but not RA) caused an inhibition of cell growth which was reversible. Some granulocytic maturation associated changes were induced by both agents: nuclear segmentation in 10-20% of cells, B43.4 antigen positivity. In contrast, several other markers were not induced: nucleoli persisted in almost 100% cells including the segmented ones, the nuclear membrane regions did not stain with Victoria blue B (which stains these regions in normal neutrophils), the expression of other antigens of mature neutrophils was also not induced. Maturation asynchrony and non-physiological segmentation of round and oval nuclei were observed. Examination of nucleoli on a single cell level revealed a reversible decrease of pre-rRNA synthesis in 28-48% of the induced cells. These results indicate that terminal differentiation did not occur and confirm the dissociation in induction of various differentiation markers.

Antigens, Surface↗
PubMed1987

Derivatives of benzo(c)fluorene. XXIV. Cytostatic effect of benfluron on human leukemic cells in vitro.

Benfluoron (BF), a new cytostatic drug, synthesized at the Institute of Pharmacy and Biochemistry in Prague, was tested for its cytostatic and cytotoxic effect. The concentrations of BF ranging from 0.1-2 micrograms/ml had a significant cytostatic effect on nine stabilized human leukemic cell lines. This effect was demonstrated by cell counts, cell viability and 3H-thymidine incorporation. The BF concentrations of 2 micrograms/ml and higher were considerably cytostatic causing the cell death and cell degradation. The effect of BF on the cell growth being irreversible could not be eliminated by washing the cells and recultivating them in fresh medium. The BF concentrations halving the total number of viable cells (EC50 value) induced a higher cytotoxicity in lymphoblastoid cell lines then in myeloid ones. BF did not influence the binding of monoclonal antibodies with membrane markers of different cell subpopulations. Prospective application of BF as a cytostatic and immunosuppressive drug is discussed.

Antibodies, Monoclonal↗
PubMed1986

Effect of ribonucleases on cell-mediated lympholysis reaction and on GM-CFC colonies in bone marrow culture.

Natural dimer of bovine seminal ribonuclease (AS RNase) suppressed markedly DNA synthesis in allogeneic mixed lymphocyte culture (MLC) of normal human lymphocytes and simultaneously inhibited induction of cytotoxic effector cells within the sensitization phase of indirect cell-mediated lympholysis (CML) reaction. The last purification step of the AS RNase isolation procedure did not increase the suppressive activity of AS RNase compared to a less purified preparation (ZS RNase), thus, the later preparation was mostly used. ZS RNase (10 micrograms/ml) caused 50% inhibition of MLC reaction whereas pancreatic ribonuclease (A RNase) was 10 times less effective. The suppressive effect of RNases added in the beginning of the sensitization phase of the CML reaction correlated with that observed in the MLC reaction. The concentrations of ZS RNase (10 micrograms/ml), A RNase (100 micrograms/ml), and additionally tested cyclosporin A (0.5 microgram/ml) resulted in nearly total abrogation of cytolysis in CML. ZS RNase added after the sensitization of effector cells did not influence their cytolytic action on target cells within the destruction phase of CML. Natural killer and killer cell activities in normal peripheral lymphocytes were not inhibited by ZS RNase at the concentration of 330 micrograms/ml. ZS RNase (20 micrograms/ml), cocultivated 1 h with normal human bone marrow cells and then washed off, enhanced formation of GM-CFC colonies in semisolid agar culture up to 200%. Simultaneously tested antilymphocyte globulin increased the number of GM-CFC colonies at the average of 128%. This stimulating effect on colony formation appeared also in bone marrow culture of patients suffering with various hematological disorders. The possibility of utilizing the preparations gained from seminal plasma in clinical bone marrow transplantation is discussed.

Antilymphocyte Serum↗
PubMed1984

Experiences with evaluation of immunosuppressive activity of alpha-globulin fraction of human blood plasma.

Presented are experiences with the determination of immunosuppressive activity of alpha globulin fraction of human blood plasma, crude (Cohn fraction IV), partially purified and so-called low-molecular isolates of these substances. For testing the following techniques were used: the lymphocytotoxic and lymphoagglutination test, rosette-inhibition tests with human lymphocytes and splenocytes of mice immunized with sheep erythrocytes, lymphocyte blastic transformation inhibition test using PHA stimulated cells as well as in the mixed culture, inhibition of haemolytic plaque formation tests, determination of GVH reactivity using regional tests in the popliteal nodes, determination of HVG reactivity by skin graft survival test, determination of the effect on formation of precipitins against soluble antigens. The best results were obtained with the determination of survival times for skin grafts and the regional test in the popliteal nodes Fairly reproducible results were obtained in experiments on influencing the humoral antibody response and inhibition of formation of precipitating antibodies. Of in vitro methods tests of inhibition of lymphocyte blastic transformation both after PHA stimulation and in the mixed culture are applicable. Fully unsuitable proved the rosette, lymphocytotoxic and lymphoagglutination tests. In vitro modification of haemolytic plaque formation tests. In vitro modification of haemolytic plaque formation test is unfit for testing the crude fractions or partially purified drugs, its in vivo modification seems more feasible provided a large panel of animals is examined.

Alpha-Globulins↗
PubMed1984

Derivatives of benzo(c)fluorene. X. Inhibitory effect of Benfluron on cellular immunity.

The effect of a cytostatic drug Benfluron-- 5(2-N,N-dimethyl-amino-ethoxy)-7-oxo-7H-benzo(c)fluorene hydrochloride was tested in mice on skin graft survival, graft-versus-host reaction (GVHR) and for mitogenic stimulation of human lymphocytes. Application of Benfluron resulted in a prolonged skin graft survival. The regional and systemic GVHR was potentially inhibited by p. o. administration of Benfluron at a dose of 100 mg/kg of body weight. Allogeneic spleen cells incubated with Benfluron at concentration of 5-20 micrograms/ml showed reduced capability to induce GVHR. PHA or PWM stimulation of lymphocytes was completely inhibited in the presence of 2 micrograms/ml Benfluron. Benfluron as a potent immunosuppressive agent can be considered useful for the prevention of GVHR.

Animals↗
PubMed1983

Differentiation of human myeloid leukemia cell lines induced by tumor-promoting phorbol ester (TPA). I. Changes of the morphology, cytochemistry and the surface differentiation antigens analyzed with monoclonal antibodies.

Human myeloid leukemia cell lines ML-1, ML-2, ML-3, promyelocytic leukemia cell line HL-60 and histiocytic lymphoma cell line U-937 were induced to differentiate by 0.5-10 ng/ml (0.8-16 nM) 12-O-tetradecanoylphorbol-13-acetate (TPA). After 48-72 h of induction, changes of the morphology, cytochemistry and of the antigenic phenotype of induced and control cells were studied using a panel of monoclonal antibodies against granulocytic, monocytic, HLA-ABC and HLA-DR antigens in indirect immunofluorescence. Cells of the TPA-treated cultures acquired morphological, cytochemical and antigenic markers of monocytes/macrophages, as surface adherence, alpha-naphthyl acetate esterase (alpha-NE) and acid phosphatase activity and the expression of monocytic antigens detected with monoclonal antibodies 63D3, FMC 17, B 44.1, B 52.1 and anti-Mol. During differentiation in vitro induced by TPA, also loss of HLA-DR antigens and diminution of antigen of cell activation were detected with antibodies L 243 and 4F2. The expression of granulocytic antigens was only slightly diminished and the expression of HLA-ABC antigens was not changed by TPA-treatment. There were differences in the percentage of cells induced to differentiate among the lines of different origin and even among the lines ML-1, ML-2 and ML-3, established from a single patient with acute myeloid leukemia. After treatment of cultures with 5 ng/ml TPA for 72 h DNA synthesis was inhibited to 60-80%.

Acid Phosphatase↗
PubMed1983

Immunosuppressive effects of bovine seminal fluid fractions with ribonuclease activity.

Using different isolation procedures (after acidification and saturation with 3 M ammonium sulphate) three fractions were isolated from bull seminal vesicle fluid and assayed for their effects on cell immunity in vitro and in vivo. Two of these preparations (ZS RNase and AS RNase) possessing a high level of ribonuclease activity at concentrations of 50 micrograms/ml showed inhibitory effects (up to 80%) on 3H-thymidine incorporation into the DNA of mitogen-or antigen-stimulated human lymphocytes. The third preparation (3M-P) possessing low ribonuclease activity showed lesser inhibitory effects. The potency of mouse spleen cells to cause regional GVH reaction was significantly decreased after preincubation of spleen cells to cause of 1 mg per AS RNase or ZS RNase whereas 3M-P was ineffective in this test. A single dose of 1 mg per mouse of ZS RNase or 3M-P administered i.p. on day 4 after skin transplantation significantly prolonged graft rejection. Both preparations at this dose potentiated the effect of cyclophosphamide on skin graft survival. All tested preparations preincubated with mouse bone marrow cells had no adverse effects on their colony-forming activity (in the spleens of irradiated mice). The possibility of utilizing the preparations with ribonuclease activity isolated from vesicle fluid in clinical bone marrow transplantation is discussed.

Animals↗
PubMed1983

Study on immunosuppressive activity of alpha-globulin fraction of human blood plasma.

Immunosuppressive activity of alpha-globulin fraction of human blood plasma (Cohn fraction IV) was studied to determine the possible clinical utilization of this fraction as such, purified, or as the starting material for isolation of the active factor. The ethanol fractions IV, isolated and waste in the production of blood derivatives were found to be directly inapplicable because of relatively low activity and poor solubility. Therefore purification experiments were done to remove the denatured proteins and lipids and to increase the specific activity, they were performed on laboratory scale using DEAE ionex chromatography, and on semipilot scale utilizing the principles of ethanol fractionation of plasma. From the viewpoint of technology, the latter procedure seems more feasible; partial purification yielded good solubility and somewhat higher specific activity. Under study are further techniques capable of eliminating the contaminating proteins with do not carry the immunosuppressively active component, as well as the effectivity of pasteurization at 60 degrees C/10 hrs., which seems indispensable with respect to inactivation of infectious hepatitis viruses and the potential clinical application. Experimental isolations of the active component have led immunosuppressively active low-molecular part, which, however, is chemically nonhomogeneous. Its subsequent subfractionation will be necessary.

Animals↗