Referate

Ohne Zusammenfassung     

Referate

Ohne Zusammenfassung     

Immunzytologische,autoradiographische und elektronenmikroskopische Untersuchungen an peripheren lymphatischen Reizformen

Zusammenfassung Bei Erkrankungen mit vermehrtem Auftreten lymphatischer Reizformen im peripheren Blut wurde die immunzytologisch bestimmte Zahl gamma-Ghaltiger peripherer Lymphozyten mit der von³H-Thymidin-markierten Lymphozyten verglichen. Bei Patienten mit Mononucleosis infectiosa Hepatitis epidemica, Morbilli und Rubeolen ergab sich eine Zunahme beider Zellpopulationen, bei Patienten mit Morbus Hodgkin wurde eine isolierte Vermehrung der³H-Thymidin-markierten Lymphozyten beobachtet. Im zeitlichen Ablauf erreichten zuerst die³H-Thymidin-markierten, dann die gamma-G-haltigen Zellen die höchsten Werte. Bei der direkten Zuordnung zeigte sich, daß bis zu einem Fünftel der IgG-haltigen Zellen³H-Thymidin-markiert waren und somit teilungsfähig, sein dürften. Eine der Zahl der IgG-haltigen Lymphozyten vergleichbare, wenn auch etwas größere Anzahl lymphoider Zellen wies bei der elektronenmikroskopischen Klassifizierung vermehrt Ergastoplasma auf.

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Summary The immunocytologically determined number of gamma-g-containing peripheral lymphocytes was compared in diseases showing an increased appearance of lymphatic irritation cells in the peripheral blood. Patients suffering from mononucleosis infectiosa hepatitis epidemica, measles and rubella showed an increase in both cell populations. Patients suffering from Morbus Hodgkin showed an isolated increase of³H-tymidine labelled lymphocytes One after another did at first the³H-thymidine labelled cells reach the highest values, then the gamma-g-containing cells. On direct coordination it could be demonstrated that the IgG-containing cells where up to one fifth³H-thymidine labelled, and consequently should be separable. A greater amount of lymphoid cells, comparable to the amount of IgG-containing cells, showed a considerable quantity of ergastoplasm in the electronmicroscopic classification.

     

Productive infection of in vitro generated haemopoietic progenitor cells from normal human adult peripheral blood with parvovirus B19: studies by morphology, immunocytochemistry, flow-cytometry and DNA-hybridization

Parvovirus B19 exerts a highly selective cytopathic effect on erythroid progenitor cells. Studies so far on the pathogenesis of B19-infection have been performed using bone marrow samples providing large amounts of erythroid progenitor cells. Extensive study, however, has been hampered by the limited access to bone marrow samples. We have designed a liquid culture method allowing the generation of large numbers of erythroid progenitor cells, initiating cultures with CD3- and CD14-poor peripheral blood mononuclear cells. Following a 12 d preincubation in liquid cultures containing recombinant human interleukin 3 (rhIl-3) and recombinant human erythropoietin (rhEpo), cells harvested from the liquid cultures were exposed to B19-containing plasma, followed by a further cultivation in liquid culture for up to 96 h. Cells expressing the CD13 and the glycophorin A (GlyA) antigens, respectively, were monitored sequentially by flow-cytometry, demonstrating a selective inhibition of GlyA-positive cells following B19-inoculation. Typical morphological changes were observed on cytocentrifuge-spots, and typical giant-cells were identified as staining for GlyA. Productive infection by B19 was demonstrable, as B19-DNA increased by about x 100 after 72 h of culture. The liquid culture method generating erythroid target cells for effective infection by B19 virus promises to be a useful and easily accessible tool for further research on B19 infection of haemopoietic cells.     

Acute parvovirus B19 infection mimicking myelodysplastic syndrome of the bone marrow

Summary A 36-year-old, previously healthy woman was referred to our institution with pancytopenia and splenomegaly for suspected acute leukemia. Bone marrow aspiration showed marked dysplastic changes, excess of blasts, and only spurious red blood cell precursors. Action was taken to prepare allogeneic bone marrow transplantation from an HLA identical sibling for myelodysplastic syndrome. Repeat cytological examination of the bone marrow revealed striking hyperplasia of the red cell line with presence of abnormal giant proerythroblasts. Acute parvovirus B19 infection was suspected and confirmed by detection of anti-B19 IgM and B19 DNA. The underlying disease for this transient aplastic crisis was a formerly unknown hereditary spherocytosis.     

CD 34 Immunotyping of blasts in myelodysplasia

Summary We studied the expression of the hematopoietic progenitor cell antigen CD 34 in six patients with refractory anemia with excess of blasts (RAEB), five patients with RAEB in transformation (RAEB-T), and seven patients with chronic myelomonocytic leukemia (CMML). Immunocytochemical labeling of bone marrow cells was performed by an indirect immunoperoxidase method with preservation of morphological details. The cells were stained with May-Grünwald-Giemsa, photographed, destained, and immunolabeled by the immunoperoxidase technique. We found 1.5±0.5% blasts and 0.8±0.4% CD 34⁺ blasts in normal bone marrow. The CD 34 positivity of blasts was 53±9%. The patients with RAEB showed 1.7±1.4% CD 34⁺ blasts. The CD 34 positivity of blasts (11.8±5.6%) was lower than in normal bone marrow. The patients with RAEB-T had a higher percentage of CD 34⁺ blasts (7.33.4) and a higher CD 34 positivity of blasts (28.2±14.6%) than patients with RAEB. The CMML patients showed a percentage of CD 34⁺ blasts and a CD 34 positivity of blasts in the range of RAEB. We found an increase of promonocytes (PMC) in 5/7 patients. In some patients the PMC were CD 34 positive. Our results indicate that the increase of blasts in REAB is related to CD 34-negative blasts. With progression to RAEB-T the percentage of CD 34-positive blasts increased. Some of the CMML patients also showed a population of CD 34-positive PMC. A clone of undifferentiated CD 34-positive cells is characteristic for patients with these types of myelodysplasia.This work was presented at the Annual Congress of the German and the Austrian Society of Hematology and Oncology, Essen, 10–13 October 1993     

Feinstruktur zirkulierender Lymphozyten unter Einwirkung von Antilymphozytenserum (ALS) bei chronischer lymphatischer Leukämie

Zusammenfassung Bei drei ALS-behandelten Patienten mit chronischer Lymphadenose wurden die zirkulierenden Lymphozyten vor und während der ALS-Gabe sowie nach Inkubation mit ALS elektronenmikroskopisch untersucht. Die Lymphozyten zeigten während der Behandlung weder qualitative noch quantitative (Einteilung in 4 Gruppen, Ausdifferenzierung) Veränderungen. An den ALS-inkubierten Lymphozyten waren verschiedenartige Schäden nachzuweisen, die mit entsprechenden Befunden anderer Untersucher verglichen werden.

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Summary Lymphocytes of three patients suffering from chronic lymphocytic leukemia were studied electromicroscopically before and during ALS-treatment and after in-vitro-incubation with ALS. During the treatment, no qualitative or quantitative changes were shown. After incubation with ALS different changes could be demonstrated. The findings are discussed.

     

Analysis of chronic lymphoid leukaemias according to FAB.

We have studied the immunophenotypic features in patients with chronic lymphoid leukaemia and investigated the suitability of classification according to guidelines of the French-American-British (FAB) group. Immunophenotyping was carried out on cytocentrifuge preparations of mononuclear blood leukocytes using the alkaline phosphatase-antialkaline phosphatase (APAAP) method. The 114 leukaemias, including 58 cases of B-chronic lymphocytic leukaemia (B-CLL), 3 Waldenstr?m's macroglobulinaemia, 6 prolymphocytic leukaemia (B-PL), 13 B-CLL/PL, 4 B-CLL of mixed cell type, 8 splenic lymphoma with villous lymphocytes (SLVL), 8 hairy cell leukaemia (HCL), two HCL variant, three leukaemic phase of follicular lymphoma, two leukaemic phase of intermediate lymphoma, two plasma cell leukaemia and two chronic T-cell leukaemia, were investigated. The 111 of 112 B-chronic lymphoid leukaemias (B-CLL + B-PL + B-CLL/PL + SLVL + HCL etc.) showed monotypic light chains. The antibody HML1 was highly specific for HCL. The antibodies CD11c and CD25 were positive in all HCL cases, but were not specific for this disease. CD5 positivity and CD22s negativity were found in most patients with B-CLL, B-CLL/PL and B-CLL of mixed type. This marker type has a limited value for differentiation from the other chronic lymphoid leukaemias. We also studied three patients with chronic lymphoid leukaemia which were not described by the FAB classification. We conclude that a study of the morphology of the leukaemic cells was the most useful basis for the diagnosis of these leukaemias, whereas immunotyping was apparently valuable only in individual cases.     

Metabolic products of microorganisms. 228. New nikkomycins produced by mutants of Streptomyces tendae

Four new dipeptidyl nikkomycins of the Z- and the X-series with a variation in the amino acid moiety of the molecule were isolated from the mutant Streptomyces tendae 901/395 and characterized. Nikkomycins Kz and Kx contain 2-amino-4-hydroxy-4-(2-pyridyl)butyric acid, and nikkomycins Oz and Ox 2-amino-4-hydroxy-4-(5-hydroxy-2-pyridyl)butyric acid. In contrast to nikkomycins Z and X, nikkomycins Kx and Ox are quite stable at alkaline pH and exhibit a lower biological activity against various test organisms. From the mutant S. tendae 901/C37, which is auxotrophic for methionine and threonine, enhanced amounts of two tripeptidyl nikkomycins, Qz and Qx, were produced which are analogues of nikkomycins J and I and contain a homoserine residue instead of glutamic acid. These nikkomycins exhibit a high pH instability.