Acute leukaemia with B- and T-lymphoid and myeloid characteristics in a patient with a myelodysplastic syndrome.
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Animals↗
Biomedical subjects
A Neubauer
Publications and source records attributed to A Neubauer.
The expression of the transferrin-receptor parallels the state of proliferation in HL-60 cells.
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Antibodies, Monoclonal↗
Flow-cytometric determination of intracellular pH, esterase activity and cell volume in human leukemic cell lines following in vitro incubation with cytostatic drugs.
A recently developed flow cytometric assay method using patient tumor cells allows the determination not only of their sensitivity to cytostatic drugs but also of biochemical and biophysical parameters after treatment, such as esterase concentration and intracellular pH of the living cells. DNA-content of the dead cells and cell volume of living and dead cells. The T-cell lines CEM, Molt4, Jurkat, the B-cell lines RPMI1788, Daudi, Raji and the promyelocytic line HL60 were incubated with: cytosine arabinoside (ara-C), L-asparaginase, daunorubicin, vincristine and prednisone for 48 h. Living cells then stained with esterase and pH-dye 1,4-diacetoxy-2,3-dicyanobenzene (ADB) and dead cells with DNA-dye propidium-iodide (PI). The esterase concentration, an index of metabolic activity, decreased in the T-cell lines under the influence of ara-C, daunorubicin and vincristine, whereas in the B-cell lines smaller changes in esterase concentration were observed (P less than 0.001). A decrease in intracellular pH was seen in the ara-C and daunorubicin-incubated cells Molt4, CEM and HL60, whereas in the B-cell lines no significant change in intracellular pH was found. In all lines except Jurkat the cell volume of the surviving cells increased under the influence of certain drugs (primarily ara-C and daunorubicin); B-cell lines showed a greater swelling than T-cell lines (P = 0.001).
Antineoplastic Agents↗
Mutations of the ras protooncogenes in chronic myelogenous leukemia: a high frequency of ras mutations in bcr/abl rearrangement-negative chronic myelogenous leukemia.
Seventy cases of chronic myelogenous leukemia (CML) were analyzed for the presence of ras mutations using polymerase chain reaction (PCR), oligonucleotide hybridization, and direct PCR sequencing. All cases had preceding cytogenetic and bcr rearrangement studies. Aberrant ras genes were detected in none of 39 patients with Philadelphia (Ph) chromosome or bcr/abl rearrangement positive chronic-phase CML and in only 1 of 18 patients in blast crisis, suggesting that ras mutations have little or no role in initiation or progression of common CML. Seven of 13, or 54% of patients with bcr/abl rearrangement negative chronic phase CML (atypical CML) harbored mutations in ras, however. This high incidence of ras mutations, together with the absence of bcr/abl rearrangement, provides evidence that atypical CML is an entity that is molecularly distinct from common CML. Moreover, the clinical characteristics and the high frequency of ras mutations suggest that atypical CML may constitute a subset of the myelodysplastic syndrome and may be best classified as a variant of chronic myelomonocytic leukemia (CMML).
Base Sequence↗
Lateral eye movements as an indication of hemispheric preference: an EEG validation study.
The hemispheric asymmetry model of lateral eye movements (LEM), hypothesizing a relation between a predominance of lateral gazes to one side with a predominance of activity in the hemisphere contralateral to the preferred direction of gaze, was tested in male, right-handed subjects. During presentation of cognitive tasks (verbal synonym tasks and spatial orienting tasks) the decrease in alpha-power from a reference period to each of four activation periods (anticipation, task-presentation, first LEM, answer) for 6 derivations from each hemisphere was calculated using the method of 'Event-related desynchronisation'. Evidence for the hypothesis was found only during the EEG-epoch one second before the answer. Discrimination between subjects with a predominance of left vs right eye movements (left- vs right-movers) was best at temporal (T3, T4) and central (C3, C4) sites. These results were significant for the entire alpha-band (8-12 Hz) and--even more pronounced--for the alpha-2-band (10-12 Hz). The alpha-1-band (8-10 Hz) did not produce significant differences. Comparing the two types of tasks, synonym tasks yielded results which were more congruent with the hypothesis. The results suggest that lateral eye movements indicate individual differences in hemispheric EEG-asymmetries under certain experimental conditions (task, segment of EEG-analysis, site, frequency band).
Adult↗
[Multicenter prospective risk-adapted study on the therapy of non-Hodgkin's lymphomas of high malignancy. Use of COP-BLAM/IMVP-16 and randomized adjuvant radiotherapy--study concept and preliminary results].
In a multicenter prospective randomized therapeutic trial in advanced (stage II-IV disease, Ann Arbor classification) high-grade malignant non-Hodgkin's lymphomas (NHL, Kiel classification) a sequential combination of the COP-BLAM (5 cycles) and the IMVP-16 (2 cycles) protocols was employed. Response was first determined after 2-3 cycles. In a response-adapted manner the therapy was immediately switched to IMVP-16 if only a partial remission or no response was obtained as evidenced by the first restaging. The aim of the study is the investigation of the efficiency of this concept to induce stable remissions. In an additional randomized trial, involving all patients reaching complete remissions after chemotherapy (second restaging), the prognostic relevance of adjuvant radiotherapy as compared to therapy-free follow-up is evaluated. Eighty percent of the 191 recruited qualified patients have so far become evaluable. Complete clinical remissions were achieved in 76/148 (51%) of the patients up to the first, in 52/85 (61%) of the patients up to the second restaging. Only in a few cases did the expected toxicity of intensive polychemotherapy reach WHO grade 3-4, including nausea and diarrhea, infections, septic complications, myelotoxicity, and stomatitis. Four of the 29 deaths recorded so far occurred in complete remission due to treatment-related complications, whereas 22/29 (76%) died in progression and 3 of unrelated causes.
Antineoplastic Combined Chemotherapy Protocols↗
Evidence of cross-reactivity of 'carcinoma-specific' KC4 monoclonal antibody with activated mesothelial cells and phytohemagglutinin-stimulated lymphocytes.
Carcinoma-specific antibodies would be a useful tool in immunocytology of serous effusions. We tested the carcinoma-'specific' monoclonal antibody KC4 with cells obtained from pleural effusions evaluated by thoracoscopy and pleural biopsies. KC4 reacted most strongly with carcinoma cells. However, activated mesothelial cells also expressed this marker strongly. Lymphocytes stimulated with phytohemagglutinin were also stained by KC4. Thus KC4 appears to detect a proliferation antigen rather than a carcinoma-specific antigen.
Antibodies, Monoclonal↗
[Prospective comparison of the diagnostic value of cytology and immunocytology in pleural effusion studied by thoracoscopy and biopsy].
Two issues have been elaborated: the value of immunocytochemistry in the diagnosis of pleural effusions, and the reactivity of the investigated antibodies with different classes of cells in pleural effusions. Effusions of unknown origin from 38 patients were investigated using thoracoscopy, pleural biopsies, conventional cytology, and immunocytochemistry. The following antibodies were used: those monoclonal against various leukocyte antigens, macrophage antigens, epithelial membrane antigen (EMA), various cytoskeleton antigens, and melanoma antigens; those polyclonal against CEA and ferritin. All of the techniques used showed 18 patients (48%) as having a tumor-cell negative effusion. A pleural tumor with a malignant effusion showed in 13 patients (34%); in 12 of these immunocytochemistry also revealed tumor cells. Seven patients (18%) had a tumor of the pleura with a tumor-cell negative effusion; in 2 of these immunocytochemistry revealed a tumor-cell positive effusion. There was no difference with regard to the number of NK cells in patients with inflammation of the pleura and negative cytology and patients with tumor of the pleura and malignant effusion (3% vs 4.5%). Tumor cells were mainly stained by EMA, cytokeratin, and CEA. CEA was the only antibody to be tumor-cell specific, while EMA and cytokeratin were expressed by mesothelial cells also. The antibody against ferritin was a significant marker for mesothelial cells.
Antibodies, Monoclonal↗
[HIV infection caused by needle injury].
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Acquired Immunodeficiency Syndrome↗
Cytostatic drug testing in human leukemias by means of multiparametric flow cytometry.
Human bone marrow cells from 20 patients as well as the permanent human B-cell lines RPMI 1788, Raji, Daudi, T-cell lines Molt, CEM, Jurkat and the promyelocytic line HL 60 were assayed by means of a newly developed in vitro flow cytometric cytostatic drug assay. The cells were exposed to cytosine-arabinoside, L-asparaginase, daunorubicin, prednisone or vincristine. Surviving cells were stained after an incubation period of 2 to 7 days with esterase and pH-indicator dye ADB (1,4-diacetoxy-2,3-dicyanobenzene), dead cells with DNA-dye PI (propidium iodide). Dose-response curves were established using percent surviving cells. It was possible to evaluate bone marrow samples from 16 out of 20 patients. Seven samples were leukemic (acute myeloid leukemia (AML) n = 6, Non-Hodgkin's Lymphoma (NHL) n = 1). Nine samples were from patients either in complete remission or with benign diseases. Daunorubicin and cytosine-arabinoside were cytotoxic in both groups, whereas vincristine was effective mainly in the leukemic group (p less than 0.05). There was significant heterogeneity in the reactivity of AML-marrow cells from different patients to different drugs. The cell lines exhibited different patterns of sensitivity. Vincristine arrested cells in G2/M-phase, cytosine-arabinoside caused an increase of cells in the S-phase.
Antineoplastic Agents↗
In vitro bioassay of LH in human serum: the rat interstitial cell testosterone (RICT) assay.
A sensitive bioassay for serum LH and hCG has been developed by utilization of the testosterone response of collagenase-dispersed rat Leydig cells to gonadotropic stimulation in vitro. Testosterone production by dispersed interstitial cells is stimulated by human, ovine, bovine, porcine, rat and rabbit LH, and by hCG and pregnant mare serum gonadotropin. The rat testis interstitial cell assay gives parallel dose-response curves for all steroidogenic gonadotropins tested, and thus permits cross-species comparison of the intrinsic biological activities of native and modified gonadotropins. The sensitivity of the rat interstitial cell bioassay is equal to or higher than that of radioimmunoassay, with detection limits of 50 muU for hMG and 20 muU for hCG. The optimum conditions for bioassay of serum gonadotropins were provided by incubation of dispersed interstitial cells in the presence of 1-methyl 3-isobutyl xanthine with the addition of gonadotropin-free serum or 5% BSA to ensure a constant proportion of serum protein in all assay samples and standards. Assays performed under these conditions gave parallel dose-response curves and identical maximum responses to both standards and serum samples containing endogenous LH or hCG. All responses to human gonadotropin standards and serum samples in the bioassay were abolished by incubation in the presence of antisera to LH or hCG. This sensitive and specific method permits bioassay of basal levels of LH in male and female serum, and of the higher gonadotropin levels in postmenopausal and pregnant subjects after appropriate dilution of the serum samples. For normal men and women, serum samples of 25-100 mul are employed, while volumes of 1-20 mul are adequate for assay of LH in postmenopausal or hypogonadal subjects. Serum LH values in hypopituitary and oral contraceptive-treated subjects were usually undetectable, while LH levels in normal subjects were always measurable with good precision. The within-assay coefficient of variation for measurement of a normal male plasma pool (30 mU/ml) was +/- 10%, and the between-assay variation was +/- 15%. The precision of the assay (lambda) was 0.035 +/- 0.015 (n = 72). In 42 normal females, the bio: immuno (B:I) ratio for serum LH was 1.20 +/- 0.40 (SD), and no consistent change in ratio was observed throughout the menstrual cycle. By contrast, significantly higher B:I ratios were observed in normal males (2.5 +/- 0.4), postmenopausal females (2.6 +/- 0.6) and patients with Turner's syndrome (2.6 +/- 0.6). These findings suggest that the LH circulating in normal female plasma is similar in biological and immunological activity to hMG, whereas the LH present in normal male plasma and in states of increased gonadotropin secretion has a relatively higher biological activity. The rat interstitial cell bioassay provides for the first time a practical and precise assay for measurement of the biologically active LH levels in serum of normal biologically active LH levels in serum of normal human subjects.
Animals↗
[On the occurrence of a strain of cells with 55 chromosomes and diplo-Philadelphia chromosome before the terminal blast crisis of a chronic myeloid leukemia].
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Adult↗
Outcome of peripheral blood stem cell mobilization in advanced phases of CML is dependent on the type of chemotherapy applied.
High-dose chemotherapy with autologous transplantation of in vivo purged PBSC is a novel investigational approach to treating chronic myelogenous leukemia (CML) patients not responsive to conventional therapy with interferon-alpha (IFN-alpha) and not eligible for allogeneic transplantation. PBSC mobilization using either '5+2/7+3'-type chemotherapy or 'mini-ICE/ ICE' chemotherapy was investigated in 43 patients with advanced phases of Philadelphia (Ph)-positive CML. Thirty patients were in late chronic phase (>12 months post diagnosis) and 13 patients in accelerated phase (AP) or blast crisis (BC). Contamination with Ph-positive cells was evaluated in harvests from 37/43 patients. The outcome of PBSC mobilization was dependent on the type of chemotherapy administered: a complete or major cytogenetic response (<35% Ph-positive metaphases) in leukapheresis collections was obtained in ten of 15 patients treated with 'mini-ICE/ICE' but in only three of 28 patients treated with '5 + 2/7 + 3' chemotherapy. One patient (1/43) in blast crisis died during mobilization therapy (2%). Twenty-five patients underwent PBSC transplantation and all of them engrafted successfully. Transplantation-related mortality was 0%. The data show that in advanced phases of CML the chance of harvesting Ph-negative peripheral blood stem cells depends on the type of chemotherapy used for mobilization.
Adult↗
Efficient generation and characterization of tumor cell subclones with different adhesion pathways involved in cell lysis.
BACKGROUND: Specific tumor cell recognition is required for optimal tumor directed therapy. Lymphokine activated killer (LAK) cells are able to recognize tumor targets specifically because LAK cells can distinguish between normal and tumor cells. This study was aimed at analyzing receptor molecules on tumor cells and their counter-receptor molecules on LAK cells. Cell lines which differ in the pathway by which LAK cell lysis is mediated are important for an analysis of receptor molecules. METHODS: We adapted a novel method for efficient production of K562 clones in order to analyze the mechanisms by which target and effector receptor molecules mediate different LAK cell interactions. K562 cells were exposed to ethyl methanesulfonate (EMS) for mutagenization and addition of two rounds of irradiated LAK cells. Prior data demonstrated that irradiation does not effect cytolysis. Surviving cells were plated in methylcellulose and single colonies were obtained after ten days. Cells were washed, resuspended in medium, expanded and tested as targets in a 51Cr-release assay. RESULTS: With this procedure a variety of clones could be generated easily and time-savingly. All twelve clones expressed the bcr/abl transcript, as determined by PCR, and were sensitive to LAK cell lysis. However, cell blockage studies revealed that K562 clones were generated with LAK cell recognition differing from parental K562. Antibody blockage showed that lysis of clone 5 (LEF 5) is partially mediated via the LFA-1/ICAM-1 pathway. ICAM-1 expression of this clone was similar to expression on K562, as determined by flow cytometry. CONCLUSIONS: These clones are of great value for studying the receptor molecules involved in LAK--tumor cell interactions.
Cell Adhesion↗
Increase of cytotoxic sensitivity of primary human melanoma cells transfected with the interleukin-7 gene to autologous and allogeneic immunologic effector cells.
Patients with metastatic melanoma have a very poor prognosis. In many cases, the tumor recurs after surgical excision. Therefore, it might be beneficial for cancer patients to induce an immune attack against the tumor by inserting a cytokine gene into the tumor cells. Here, 14 primary cell cultures could be established from 45 patients with malignant melanoma. Primary cell cultures were transfected via electroporation with the gene encoding for human interleukin-7 (IL-7). Transfection resulted in the production of biologically active IL-7 with an average of 850 pg/mL per 10(6) cells per 24 hours. Irradiation with 10,000 cGy, which inhibited tumor cell growth in vitro, increased the amount of released IL-7 to an average amount of 1050 pg/mL per 10(6) cells per 24 hours. No significant differences in the phenotype were observed in the IL-7-transfected cells compared with nontransfected cells. The expression of HLA class I and II, ICAM-1, and of a melanoma-associated antigen remained unaltered. Transfection with IL-7 had no significant effect on the proliferation of melanoma cells as measured in a MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. There was no significant change in the cytokine profile after transfection or irradiation of the cells, but one cell culture expressed a high amount of IL-6 (about 2 ng/mL). IL-6 was expressed in nontransfected cells and was not altered by transfection. Interestingly, transfected cells from primary melanoma cultures possessed a higher sensitivity to immunologic effector cells compared with nontransfected cells. This was true for allogeneic as well as autologous melanoma cells. Our results show the feasibility of a gene transfer into primary human melanoma cells, different from retroviral transduction. IL-7-transfected cells might be of value in vaccination protocols for melanoma patients.
Cytotoxicity, Immunologic↗