Human leukemic lymphocytes--biochemical parameters of the altered differentiation status.
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Biomedical subjects
Publications and source records attributed to M Garbrecht.
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Normal lymphocytes and lymphocytes from patients with low-grade malignant non-Hodgkin lymphoma were isolated from blood by a Percoll gradient procedure. Absence of cell proliferation in both cell types was indicated by very low [3H]thymidine incorporation rates. Determination of endogenous protein-bound single ADP-ribose residues by a radioimmunoassay revealed that the leukemic cells had 2.5-times lower levels of the NH2OH-sensitive and a 4-fold lower amount of NH2OH-resistant ADP-ribose . protein conjugate subfractions, respectively, than normal lymphocytes. By contrast, "total" ADP-ribose transferase activity, as measured in homogenates or permeabilized cells in the presence of DNase, was two-times higher in leukemic cells, whereas activity determined in permeabilized cells in the absence of added DNase was practically identical in both cell types. The apparent discrepancy between ADP-ribose transferase activity and endogenous levels of protein-bound single ADP-ribose residues may be explained in part by an enzyme inhibitor present in normal human lymphocytes. NAD + NADH levels were decreased 2.5-fold in the leukemic cells. This decrease, however, does not explain the reduced levels of mono(ADP-ribose) . protein conjugates since the ratio of protein-bound single ADP-ribose residues to NAD is distinctly different in leukemic lymphocytes compared to normal lymphocytes.
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A case of Ph1-positive chronic granulocytic leukemia in described in which a terminal monocytic crisis developed. The case still was Ph1-positive at the time of the monocyte crisis. Bone marrow smears contained promyelocytes and early granulocytes, but most of the cells belonged to the monocyte series as cytochemistry disclosed. No blast cells could be detected. Cytochemically, transitions between promyelocytes and promonocytes could be demonstrated. From these findings and those from the literature it is concluded that the monocytes of the terminal crisis in our case belonged to the leukemic cell strain and that these monocytes developed from non-specific promyelocytes like granulocytes. Our observation does not provide evidence for the existence of a separate monoblast which would implicate disparity of granulocytopoiesis and monocytopoiesis already at the level of undifferentiated blast cells. This, in turn, would be equal to a re-introduction of a modified polyphyletic theory of white blood cell formation.
Fourty-three of 61 patients suffering from low-grade malignant non-Hodgkin's lymphomas according to the Kiel classification were subjected to an intermittent combination chemotherapy with chlorambucil and prednisone. In 12 of 35 evaluable patients a complete remission, in 14 of 35 a partial remission could be achieved. The mean remission time of patients with a complete remission exceeds 16 months, that of patients with a partial remission amounts to more than 8 months. In 9 of 35 cases a remission could not be achieved. As 10 of the 12 patients in whom a complete remission could be obtained are alive, the median survival time in cases of complete remission cannot yet be determined. The hematological toxicity was very low, nausea and lack of appetite were observed in 9 of 35 patients. Three of 35 treated and evaluable patients died, viz., 1 patient with a centroblastic-centrocytic lymphoma and 1 patient with a chronic lymphocytic leukemia died from the sequelae of progression during the chemotherapy of second choice according to the COP regimen. The third patient with a centroblastic-centrocytic lymphoma died from the sequelae of an intrahepatic cholestatic icterus, also during the chemotherapy of second choice according to the COP regimen. In the last-mentioned case, autopsy also confirmed a continuous complete remission of the centroblastic-centrocytic lymphoma.
The qualitative and quantitative analysis by high performance liquid chromatography of the normal and modiefied nucleobases excreted in urine represents a new and versatile tool, especially in oncology. The excretion of 2 normal (adenine, guanine) and 4 modified nucleobases (methylated guanine derivatives) in urine was measured by cation exchange LC. All chronic myeloproliferative syndromes showed highly elevated total excretion values of all determined nucleobases, the "pattern" being characteristic with N2, N2-dimethylguanine most prominent (up to 29.8 S.D. above the pertinent normal value). A follow-up study of a case of CML with two episodes of extreme leukocytosis showed a correlation of the nucleobases excretion with the number of leukocytes. Thus, a method has been established which permits the assessment of myeloproliferation and probably therapy effects.
Rebound regression occurred in 13 of 42 females (30.9% with metastasising carcinoma of the breast in the postmenopausal period in whom additional hormonal treatment with ethinyl oestradiol had brought about a remission. Duration of the rebound response ranged from 3 to 44 months (average 11.2 months; mean 8 months). There was a significantly longer duration of remission (18.7 months compared with 12.0 months; P = 0.01) in patients with than those without a rebound response. There was also a significant correlation between the duration of the preceding remission under additional hormonal treatment and the duration of the subsequent rebound response (P less than 0.01). On the other hand, there was a negative but not significant correlation between the free interval duration which preceded the metastasisation and the duration of the rebound regression.
Various ventricular functions, as obtained by echocardiography (M-mode), were measured on 14 patients with solid tumours receiving adriamycin (mean total dosage 330 +/- 188 mg/m2 body surface area), the results being compared with those on 14 normal subjects of similar ages. There were statistically highly significant differences between the two groups. In those receiving the drug there was (1) a decrease in the percentage shortening fraction and (2) a reduction in the maximal velocity of endocardial circumferential fibre shortening, as expression of reduced contractility. The abnormal filling was characterized by a marked decrease in maximal velocity of endocardial fibre stretch, diminution of diameter increase during the rapid filling phase, and a shortened duration of the rapid filling phase. There was no electrocardiographic or radiological evidence of myocardial damage. The findings suggest that it will be possible by serial echocardiography to recognize a higher tolerance to the drug in some individuals and, at the same time, reveal abnormal contractility and filling due to adriamycin before irreversible heart failure has occurred.
Carcinoembryonic antigen (CEA) was determined in 72 patients with metastatic breast cancer under various kinds of treatment. In 53 cases, CEA levels were elevated. They correlated with free interval, metastatic spreading, alkaline phosphatase (AP) and gamma-glutamyltranspeptidase (gamma-GT). Serial CEA determinations in 50 of the 72 patients revealed increased CEA levels in connexion with progressive tumour growth and decreased CEA levels in connexion with response to treatment. CEA may therefore be useful in helping to assess the extension of the tumour and the effect of treatment in patients with matastatic breast cancer.
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Splenectomy was perfomed in three patients with typical chronic myeloid leukaemia after a compensated stage had been reached with busulphan. Marked pancytopenia as a result of hypersplenism occurred in one patient 14 months after the end of the primary induction, and was successfully treated by splenectomy after radiotherapy of the spleen had failed. Splenectomy was performed without complication, and the histological appearance of the spleen showed myeloid infiltrates while there was none in the liver biopsy. After another 18 months the patient remained compensated, without busulphan administration. In the two other patients splenectomy was performed in the recompensated stage, one month and five years, respectively, after the primary induction had been concluded. In these two patients, too, there was myeloid infiltration of the spleen. In both, mild leucocytosis with shift to the left as far as promyelocytes occurred six and two weeks, respectively, after splenectomy and busulphan was started again. Twelve and ten months, respectively, after splenectomy they are both in the compensated stage. The importance of splenectomy in the early phase lies in the possibility of delaying blast crisis and avoiding myelofibrosis, as well as in the prevention of complications in the later stages.
Two of seventy patients with Hodgkin's disease who had exploratory laparotomy with liver biopsy and splenectomy were found to have extensive epithelioid-cell granulomas in liver and spleen. The coincidence of malignant tumour and epithelioid-cell granulomas of liver and spleen without lymph-node involvement is unusual. Knowing of the coincidence of Hodgkin's disease and such granulomas is clinically important because extensive diagnostic measures, including biopsy, are necessary in the differential diagnosis, and the patient's prognosis will depend on the therapeutic measures taken.
Specific activities are determined of two functional fractions of alpha-amanitin sensitive DNA-dependent RNA polymerases in nucleic from human normal and chronic lymphocytic leukemia lymphocytes. Specific activity of "free" RNA polymerase in CLL corresponds ot 0.133 pmoles (3H)-UMP/10(6) cells as compared to 0.209 in normals. Activities of the "engaged" enzymes are 0.139 in CLL and 0.132 in normals. "Free" enzymes in NL and CLL are completely inhibited by 400 ng/ml Rifamycin AF/013, while the "engaged" enzymes exhibit 70% of their original activity. 1.0 ng/ml alpha-amanitin suppress 50% of the activity of the "free" enzyme in CLL. The "free" enzyme in NL and the "engaged" enzymes in NL and CLL do not show any residual activity in the presence of 1.0 ng/ml alpha-amanitin.
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Acridine dyes inhibit the incorporation of 3H-thymidine and 3H-uridine in intact cells to the same extent as Actinomycin D. In contrast to Actinomycin D, RNA synthesis by DNA - dependent RNA polymerase in a cell-free system is inhibited at lo2 higher concentrations of acridine dyes, only. Possible differential effects on the cell membrane resulting in decreased intracellular pools of uridine and thymidine are discussed.
Specific Activities of DNA-dependent RNA polymerases A and B have been determined in nuclei from leukocytes in acute and chronic leukemia. Enzyme activities, dependent on exogenous DNA template, were determined in homogenates of nuclei from isolated mononuclear cells or from isolated granulocytes. Activities of polymerases A and B have been found significantly elevated in homogenates of nuclei from mononuclear cells in acute myelocytic leukemia, while they were found subnormal in corresponding cell fractions from chronic myelocytic leukemia and chronic lymphatic leukemia. During cytostatic treatment polymerase activities were approaching normal values. The prognostic relevance of these data for the course of human leukemia is discussed.
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