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.

The Kiel classification of non-Hodgkin lymphomas (NHL) identifies distinct entities by morphological and immunological criteria, subdivided into those of low and high grade malignancy and of B- or T-cell subtype. Numerous clinical studies have proven the clinical and prognostic relevance of this classification scheme although the degree of malignancy alone cannot be automatically equated with prognosis. Thus, histological low grade malignancy correlates with favorable response to therapy in all, but with long term survival only in some clearly defined low grade NHL and final cure is achieved in none of them so far. High grade malignancy corresponds to the aggressive spontaneous course of these NHL but is contrasted with the mostly favorable response to therapy and the chance of cure. The immunological T-cell subtype is clearly associated with an inferior prognosis. Additional biological features of NHL, namely chromosomal aberrations and the status of clonality will contribute to a more differentiated view of NHL and will possibly prove to bear prognostic potential.

Durch die Kiel-Klassifikation wurde eine Neuordnung der Non-Hodgkin-Lymphome in distinkte, auch klinisch-prognostisch differenzier-bare Lymphom-Entitäten erreicht. Im Vergleich zu früheren Klassifikationen grenzt dieses Einteilungsschema zusätzliche Lymphom-Typen (z.B. CC-Lymphom, LP-Immunozytom) (Abkürzungen s. Text) ab, denen wiederum gesonderte klinische Krankheitsbilder ent-sprechen. Im Rahmen einer multizentrischen prospektiven Beobachtungsstudie (Rekrutierung von 1127 unbehandelten Patienten von 1975 bis 1980; Nachbeobachtungsphase bis 1985) wurden eine genauere Definition des klinischen Bildes der einzelnen Lymphom-Entitäten (u.a. Häufigkeit, Alters- und Geschlechtsverteilung, Befalls- und Ausbreitungsmuster) erarbeitet sowie der Erfolg radio-und/oder chemotherapeutischer Maßnahmen überprüft. In der Gruppe der NHL von niedrigem Malignitätsgrad (69,4% der Fälle) lag initial das streng lokalisierte Stadium I/IE der Ann-Arbor-Klassifi-kation bei 1,5-8%, in der Gruppe der hochmalignen NHL (30,2% der Fälle) bei 8-17% der Patienten vor. Durch alleinige lokoregionale Radiotherapie konnte bei 86-89% (CB- und IB-Lymphome) bzw. 100% (LP-Immunozytom, CB-CC- und CC-Lymphome) dieser Patienten eine Vollremission erreicht werden; lediglich die CC- und IB-Lymphome ließen ein relevantes Rezidivrisiko (40 bzw. 50%) erkennen. Eine totallymphatische Radiotherapie führte im Stadium III des CB-CC-Lymphoms noch in etwa der Hälfte der Fälle zu anhaltenden Vollremissionen. Die Überlebenswahrscheinlichkeiten der mit verschiedenen Formen der Chemotherapie behandelten Patienten in den initialen Stadien III und IV spiegeln grundsätzliche prognostische Merkmale der einzelnen Lymphom-Entitäten wider. So weist der kontinuierliche Abfall der Überlebenskurven bei den NHL von niedrigem Malignitätsgrad auf die unabhängig von den bisherigen Therapiestrategien langfristig geringe Beeinflußbarkeit dieser Erkrankungen hin. Bei den hochmalignen NHL unterstreicht der initial steile Abfall der Überlebenskurven mit nachfolgender Plateaubildung die Bedeutung des Erreichens einer Vollremission und die daraus resultierende Heilungschance. Durch uni- und multivariate Analyse der Daten konnten für die verschiedenen Lymphom-Entitäten charakteristische Muster von Risikofaktoren identifiziert werden.

Two patients suffering from proliferation of Tγ cells exhibited uncommon clinical features, such as activation of intravascular coagulation after low dose irradiation of the enlarged spleen in one patient and isolated neutropenia in the other patient. While the malignant nature of the disease was doubtless in one patient, cell proliferation in the other patient was more likely reactive. In addition to T cell determinants the proliferating cells expressed a monocytic antigen. They did not suppress B-lymphocyte differentiation into plasma cells. In contrast the proliferating cells, especially in one patient, acted as potent effectors in NK and ADCC using melanoma and MOLT 4 target cells. Erythrophagocytosis by Tγ cells was seen in one patient. The data suggest that subsets of Tγ cells are related to the monocytic lineage and that these cells can mediate both NKA and ADCC and partly can develop phagocytic activity.

A patient with chronic leukemia exhibited uncommon clinical features, such as hypergammaglobulinemia and activation of intravascular coagulation after low-dose irradiation of the enlarged spleen. By light and electron microscopy, the leukemic cells resembled large granular lymphocytes. The following markers were simultaneously expressed on their surface: receptors for sheep erythrocytes and the Fc part of IgG; common T-cell antigens as revealed by a heteroantiserum (HuTLA) and monoclonal antibodies (OKT3, T411); antigens shared by cytotoxic/suppressor T cells (OKT8, T811) as well as myelomonocytic antigens defined by the OKM1 and M522 monoclonal antibodies. The leukemic cells showed both spontaneous (NK) and antibody-dependent (ADCC) cytotoxicity, but they did not suppress B-cell differentiation in vitro.

136 patients suffering from ALL were subdivided into 5 subtypes (C-ALL, C/T-ALL, pre-T-ALL, B-ALL) according to rosetting tests and using specific antisera directed against membrane antigens. In addition, leukaemic blasts of all patients were investigated according to morphological and cytochemical criteria. In APh and ANAE, indices and the percentages of cases showing a granular staining pattern were high in pre-T- and in T-ALL, but low in C/T- and in C-ALL. PAS-staining, conversely, was more pronounced in C/T- and C-ALL. APh proved to be more discriminative for recognition of the T- and pre-T-ALL subgroups than ANAE, but ANAE-cytochemistry may be useful to detect contaminating normal T-lymphocytes in ALL. Receptors for C3 were more frequent in C- and in T-ALL than in C/T- and in pre-T-ALL, receptors for Fc were distributed equally among all subtypes. Positively of C3- and Fc-receptors was not correlated with cytochemical results. Morphological criteria were not sufficient for subclassification of ALL; the combination of APh- and PAS-staining, however, is valuable to differentiate between C-subgroups and T-subgroups.

Zur Charakterisierung von Leukämie- und Lymphomzellen werden neben immunologischen in zunehmendem Maße auch biochemische Marker, insbesondere Enzyme, herangezogen. Ein Enzym, welches in diesem Zusammenhang interessant geworden ist, ist die membrangebundene 5′-Nukleotidase (5′-N), ein sog. Ektoenzym, dessen aktives Zentrum nach der Zellaußenseite gerichtet ist. Die normale physiologische Rolle dieses Enzyms ist noch recht umstritten und weitgehend ungeklärt. Durch histochemische Untersuchungen (Silber et al. 1975) war schon länger bekannt, daß die Lymphozyten des peripheren Bluts in eine 5′-N-positive und eine 5′-N-negative Subpopulation zerfallen, welche nicht identisch sind mit T-oder B-Zellen. Auch Beobachtungen an malignen Zellen waren bereits beschrieben, insbesondere das meist drastische Absinken der 5′-N-Aktivität der Lymphozyten bei der CLL (Silber et al. 1975) und Rosetten-positiven Formen der ALL (Reaman et al. 1979). Im Zusammenhang mit Studien zur Biochemie des menschlichen Enzyms wird in diesem Beitrag die Frage der 5′-N-Aktivität bei Leukämien und Lymphomen an einem größeren Patientengut noch einmal behandelt.

The usefulness of cytochemical tests (APh and ANAE) to replace or to supplement membrane markers in subclassification of normal and malignant lymphatic cells was investigated. Material: normal lymphocytes subfractionated by rosetting and centrifugation, and in M. Hodgkin and CLL; lymphoblastoid cell lines; malignant lymphatic cells in different types of lymphatic leukemias. In normal human blood, T-lymphocytes are marked by a distinct “dot-like” ANAE-reactivity which is somewhat less pronounced in the small (11%) subgroup of Fc-IgG-receptor positive T-lymphocytes; B-lymphocytes are negative or finely granular positive. Lymphoblastoid cell lines of B- and of T-type are ANAE- and APh-positive. In some lymphatic malignancies, a characteristic pattern of activity of APh or of ANAE may support the diagnosis. The value of ANAE-cytochemistry is highly estimated for the quantitative determination of the percentage of normal T-lymphocytes in lymphatic leukemias, immunological disorders, and during immunosuppressive therapy.

Die Proteinsynthese in Non-Hodgkin-Lymphomen (NHL) ist bisher wenig untersucht. Sie ist jedoch von besonderem Interesse, wenn man sich drei Eigenschaften dieser Tumoren vergegenwärtigt [3]. Einmal sind 80% der NHL B-Zell-Lymphome, d. h., sie sind dadurch definiert, daß sie Immunglobuline (Ig) synthetisieren. Zweitens entwickeln sich diese Tumoren nach den heutigen Vorstellungen von bestimmten Differenzierungsstufen der Lymphopoese, und diese Differenzierung geht mit wohl definierten Veränderungen innerhalb des Proteinsyntheseapparates und der Ig-Synthese einher. Drittens handelt es sich um eine Gruppe von Tumoren mit unterschiedlicher Proliferationsrate, durch die die Prognose bzw. ihr Malignitätsgrad bestimmt wird. Ausgehend von diesen Eigenschaften haben wir uns für drei Fragen interessiert: 1. Gibt es eine Korrelation zwischen Translationsparametern und klinisch bzw. pathologisch-anatomisch definiertem Malignitätsgrad? 2. Wie verhält sich der Proteinsyntheseapparat in den morphologisch und immunologisch unterschiedlich differenzierten Lymphomen? Bleibt er gleich oder ändert er sich während der Zelldifferenzierung? 3. Ist in den verschiedenen NHL auch biochemisch eine Synthese von Ig, d. h. eine Translation von Ig-Messenger-RNA nachweisbar? Wie hoch ist ihr Anteil an der Gesamtproteinsynthese und bestehen Korrelationen zum Differenzierungsgrad?

Die Klassifizierung nicht-myeloisch oder -monozytär differenzierter Leukämien bereitet Schwierigkeiten, da typische morphologische Reifungsmerkmale zumeist nicht vorhan-den sind. Die Anwendung immunologischer Methoden zur Diagnostik und Klassifizierung derartiger Leukosen (lymphatische, „undifferenzierte“) erscheint aus mehreren Gründen erfolgversprechend. In experimentellen Untersuchungen bei verschiedenen Spezies hat es sich erwiesen, daß morphologisch nicht unterscheidbare Zellen, wie z. B. die Lymphozyten, durch die Bestimmung ihrer Membraneigenschaft recht präzise in Subpopulationen unterteilt werden können, die zu Herkunft und Funktion korrelieren. Hierzu definierte Membranmarker (z. B. membrangebundenes Immunglobulin als spezifischer B-Zellmarker) und geeignete Nachweisverfahren ließen sich zumeist auf das menschliche System übertragen. Die Anwendung dieser Techniken zu diagnostischen Z wecken konnte sich jüngst als „Immundiagnostik von Leukiimien und Lymphomen“ etablieren und erbrachte bereits wesentliche neue Aspekte in Diagnostik, Pathogenese und Klassifizierung von Hämoblastosen [20].

In order to explore their differentiation capacity peripheral blasts from 4 children with acute lymphoblastic leukemia of the common type (cALL) as well as cells of the Reh line were cultured in diffusion chambers implanted intraperitoneally into preirradiated CBA mice (8 gy). At various times after initiation of the culture, changes in the membrane marker profile were studied by investigation of surface antigens and rosetting phenomena. In the course of the culture, the cells of two patients, T. H. and I.G., developed surface immunoglobulin (sIg) in combination with the CALL antigen (cALLA). Simultaneously, the cells of I.G. containing cytoplasmic Ig prior to cultivation acquired a receptor for mouse erythrocytes (EM). Besides quite a few cells developing the T-cell antigen, the majority of the cell yield in patient V.M. expressed sIg alone or together with the cALLA. Comparable results together with an increasing portion of mouse rosette forming cells were obtained in the fourth patient, C.M. In case of the Reh-line the cells developed the T-cell antigen, and in one of two experiments they further acquired a receptor for sheep erythrocytes. conversely, in a second experiment and EM-receptor was detected. Our data suggest that leukemic blast cells carrying the cALLA are representative of an early developmental stage of lymphatic ontogeny.

Acute lymphatic leukaemia (ALL) in infants is a very heterogeneous disease from the immunological point of view. Phaenotyping of leukaemic cells in 138 patients via different immunmarkers has so far allowed subdivision into 6 subgroups. Clinical risk factors and correlations with the immunologically defined subgroups, as well as their significance for prognosis, are demonstrated with the help of 61 examined ALL patients. 90-95% of all acute lymphatic leukaemias can presently be identified via heterologous antisera. After corresponding absorption, these antisera show high cytotoxicity against ALL cells without displaying any aggressivity towards normal haemopoetic stem cells. The authors make therapeutic use of anti-T cell and anti-common ALL (cALL) globulin in autologous bone marrow transplantation. This method involves withdrawal of bone marrow from patients with ALL in complete remission, the marrow being then fractionated, treated with antiserum to destroy residual leukaemic cells, and then cryopreserved. The leukaemia-free bone marrow will thus be available for transplantation in relapse.

The Fc-receptor of normal human leukocytes, of CLL-cells, and of hematopoietic cell lines was demonstrated with soluble peroxidase-anti-peroxidase (PAP) complexes. In about 9% of normal lymphocytes an almost continuous, strong labeling of the cell membrane was established. Some of these lymphocytes were characterized by a peculiar uniform fine structure. The percentage of PAP-labeled monocytes was in the range of 25%, neutrophils nearly 100%, eosinophils 0%, CLL-cells 10%. Labeled portions of the membrane were interiorized from monocytes. The lymphoid cell-line Daudi established from a Burkitt's lymphoma appeared almost negative, the cell line K562 established from a myeloid leukemia in 75% of the cells strongly positive. PAP-labeling was not influenced by preincubation with trypsine or with neuraminidase; it was negative when PAP-F(ab)2 was used. Results of PAP-labeling were not always in agreement with EA-rosettes or with agg-Ig.

According to observations in six patients and published reports, malignant histiocytosis is characterized by premature multifocal proliferation of atypical histiocytes, especially in lymph-nodes, spleen, liver, bone marrow, and lung. The diagnosis can be confirmed by electron-microscopy, and enzyme as well as immunocytochemical tests. Fever, anaemia, leukopenia (with absolute reduction in T-lymphocytes), and jaundice are frequent. Immunoglobulins are normal or polyclonally increased. Malignant histiocytosis is more like monocyte leukaemia and histiocytosis X than neoplasms of the lymphatic system.