Michael Schaadt

Establishment of a malignant, epstein-barr-virus (EBV)-negative cell-line from the pleura effusion of a patient with Hodgkin's disease

The pleura effusion was obtained from a 37-year-old woman with histologically proven Hodgkins disease, nodular sclerosing type, stage IVB, primarily diagnosed in 1972. In the 4 weeks before the last admission to our institution, the patient developed a severe eosinophilia (91 ~), the total WBC amounted to 26000/ram 3. The nodal and extranodal tumor progression did not respond to a polychemotherapy (MOPP) and the patient died due to a septical complication. The post mortem showed beside a disseminated lymphnode involvement extranodal invasion of the whole left-side chest-wall including the parietal pleura.

Cytogenetic investigations in Hodgkin's disease: I. Involvement of specific chromosomes in marker formation

Cytogenetic studies were performed in five Hodgkin-derived permanently growing cell lines, as well as in bone marrow cells from two Hodgkin patients, one of which suffering from acute myeloid leukemia after Hodgkins disease. No consistent and common marker chromosome was found, but certain chromosome regions seemed to be nonrandomly involved in marker formation. These are chromosome bands 7q22, 7q32, 7q36, 2q33, 1p22, 11q21/23, 14q32, 15p12, and 21q21. The importance and significance of these chromosomal findings are discussed in reference to data from the literature.

Sister chromatid exchange in cell lines from malignant lymphomas (lymphoma lines)

SummaryThe frequency of sister chromatid exchanges (SCEs) was studied in cells from three freshly established lymphoma lines, derived from two patients with Hodgkins disease and one patient with non-Hodgkin lymphoma. These values were compared to SCE rates found in cells from two long-established lymphoma lines (Raji and BJAB) and to those recorded in control cell lines of normal human donors. The highest SCE levels were demonstrated in the freshly established lymphoma lines, the lowest SCE values separated the lymphoblastoid cell lines from healthy controls, and the older lymphoma lines Raji and BJAB presented rates in between. The influences of BUDR concentration and of the duration of BUDR treatment on the frequency of SCEs were tested. Furthermore, the dependence of the SCE rate on the time interval between establishment of the cell line and its SCE investigation was considered. The connection between elevated SCE rates and the neoplastic nature of lymphoma lines is discussed.

Correlation of chromosomal aberrations in a myeloma cell line with tumorigenicity in nude mice

SummaryA human myeloma cell line (L 363) exhibiting several markers of malignancy including a 14q+ chromosome marker was not tumorigenic in nude mice. Long-term cultivation in vitro resulted in new subclones with slight, well-defined chromosomal differences to the original line. Certain subclones turned out to have a growth advantage in vitro and became tumorigenic after transplantation in nude mice. A strong correlation between changes in the chromosome 1 constellation resulting in the threefold dose of long arm-material and the increased proliferative potential in both experimental systems was found.

[Hodgkin's disease: effect of spenectomy on the immune status (author's transl)].

In 22 untreated Hodgkins patients the following parameters were studied before and after splenectomy: the total WBC, lymphocytes, B- and T-cells and mitogenic stimulation of peripheral lymphocytes using autologous and control serum. The results were correlated to patient groups with favourable and unfavourable prognosis according to pathological stage, histology, spleen involvement and constitutional symptoms. 1. All Patients. Significant increase of the absolute number of peripheral lymphocytes and B-cells. Significant increase of the spontanous DNA synthesis in the presence of AB-control serum, but no change after mitogenic stimulation of peripheral lymphocytes. No change in the number of peripheral T-cells. 2. Patients with Favourable Prognosis. Significant increase of the absolute number of peripheral lymphocytes and B-cells. Significant increase of the spontanous DNA synthesis in the presence of AB-control-serum. Significant decrease of the T-cell function (PHA- and Con-A-stimulation in the presence of autologous serum). Significant decrease of the PHA-, Con-A- and PWM stimulation rate using control serum in the lymphocytic predominance and nodular sclerosis group. No change in the number of peripheral T-cells. 3. Patients with Unfavourable Prognosis. No change in the absolute number of B-cells, of the spontanous DNA-synthesis using autologous serum and of the PHA- and Con-A-stimulation. Significant increase of the EBV-induced blastogenesis in the mixed cellularity and lymphocytic depletion group.SummaryIn 22 untreated Hodgkins patients the following parameters were studied before and after splenectomy: the total WBC, lymphocytes, B- and T-cells and mitogenic stimulation of peripheral lymphocytes using autologous and control serum. The results were correlated to patient groups with favourable and unfavourable prognosis according to pathological stage, histology, spleen involvement and constitutional symptoms. 1. All Patients. Significant increase of the absolute number of peripheral lymphocytes and B-cells.Significant increase of the spontaneous DNA synthesis in the presence of AB-control serum, but no change after mitogenic stimulation of peripheral lymphocytes.No change in the number of peripheral T-cells. 2. Patients with Favourable Prognosis. Significant increase of the absolute number of peripheral lymphocytes and B-cells.Significant increase of the spontanous DNA synthesis in the presence of AB-control-serum.Significant decrease of the T-cell function (PHA-and Con-A-stimulation in the presence of autologous serum).Significant decrease of the PHA-, Con-A- and PWM stimulation rate using control serum in the lymphocytic predominance and nodular sclerosis group.No change in the number of peripheral T-cells. 3. Patients with Unfavourable Prognosis. No change in the absolute number of B-cells, of the spontanous DNA-synthesis using autologous serum and of the PHA- and Con-A-stimulation.Significant increase of the EBV-induced blastogenesis in the mixed cellularity and lymphocytic depletion group.ZusammenfassungBei 22 nicht behandelten Hodgkin-Patienten wurden vor und nach Splenektomie die Gesamtzahl der peripheren Leukozyten, Lymphozyten, T-Zellen und B-Zellen, sowie die in vitro Stimulation von peripheren, und zum Zeitpunkt der Splenektomie, auch der Milzlymphozyten untersucht. Diese Daten wurden in Beziehung gesetzt zu prognostisch günstigen und ungünstigen Gruppen, entsprechend Stadium, Histologie, Milzbefall und B-Symptomatik.Für das Gesamtkollektiv ergab sich keine wesentliche Änderung des Stimulationsverhaltens peripherer Lymphozyten durch die Splenektomie. Dagegen zeigte sich ein signifikanter Anstieg der spontanen DNS-Synthese peripherer Lymphozyten mit Kontrollserum. Ebenso waren Gesamtleukozyten, Lymphozyten und B-Zellen nach Splenktomie signifikant erhöht, während sich die Zahl der T-Zellen nicht veränderte.Die Zahl der peripheren B-Zellen und die spontane DNS-Syntheserate mit autologem Hodgkin-Serum stieg in denprognostisch günstigen Gruppen signifikant an, während sich diese Parameter bei denprognostisch ungünstigen Gruppen nicht wesentlich änderten. In der T-Zellfunktion (Stimulation mit PHA und Con A) zeigten die prognostisch günstigen Gruppen mit autologem Serum einen signifikanten Abfall nach Splenektomie, während in den ungünstigen Gruppen diese Tendenz nicht beobachtet wurde. In der histologisch günstigen Gruppe ergab sich zusätzlich ein signifikanter Anstieg der EBV-induzierten Blastogenese.Die absolute Zahl der T-Zellen wurde ohne Ausnahme in keiner der Gruppen durch die Splenektomie beeinflußt.Die spontane DNS-Synthese stieg in der Untersuchung mit Kontrollserum nach Splenektomie in allen Gruppen an, während bei Zusatz von autologem Hodgkin-Serum der Anstieg nur in prognostisch günstigen Gruppen beobachtet wurde.Eine pathogenetische oder prognostische Wertung der Splenektomie bei Hodgkin-Patienten ist aus den vorgelegten Ergebnissen noch nicht möglich.

Klinische Relevanz der Immunphänotypisierung maligner Lymphome

Die Charakterisierung maligner Lymphome mit immunologischen Methoden (Immunphanotypisierung) findet eine immer breitere Anwendung in der Klinik. Sie bildet nicht nur eine Basis moderner Lymphomklassifikationen (z.B. der Kiel-Klassifikation), sondern kann daruber hinaus zur Abgrenzung von Lymphomen gegenuber epithelialen und mesenchymalen Tumoren beitragen. Bei der akuten lymphatischen Leukamie hat der Nachweis bestimmter Untergruppen (c-ALL, T-ALL, B-ALL) wegen der unterschiedlichen Prognose differentialtherapeutische Bedeutung. Als weitere Anwendungsmoglichkeiten der Immunphanotypisierung in naher Zukunft sind zu erwarten: Nachweis von Rezeptoren fur bestimmte Lymphokine (z.B. fur eine Therapie mit Interleukin-2 oder Tumor-Nekrose-Faktor), Nachweis bestimmter Oberflachenantigene (Immuntherapie mit monoklonalen Antikorpern) und Nachweis immunologischer Marker fur Zytostatikaresistenzen. Entscheidend im Hinblick auf eine breitere Anwendung der Immunphanotypisierung maligner Lymphome in der Klinik sind eine Standardisierung der verwendeten Reagenzien, eine Vereinfachung der Logistik und eine deutliche Reduktion der Kosten.