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Experiences gathered while exploring the usefulness of intravenous immunoglobulin for children with idiopathic (immune) thrombocytopenic purpura (ITP) are reviewed in view of further investigations characterizing the effects of IgG i.v. in other immune diseases without detectable antibody deficiency. The most pertinent factors to be considered are i) the heterogeneity of an immune disease investigated; ii) the criteria used to evaluate the effects of the IgG therapy; iii) the IgG preparations used and i.v. dose-fractionation. Controlled, prospective clinical trials will be required to further explore the practical usefulness of IgG i.v.

One hundred and eight children with acute lymphocytic leukemia (ALL) were admitted to a prospective therapeutic regime. Remission induction was achieved in 94% of the cases with vincristine, L-asparaginase, adriamycine and prednisone. One hundred and one patients received three intermediate dose methotrexate (MTX) infusions combined with intrathecal MTX, followed by L-asparaginase 24 h later. High risk (HR) patients (n=50) were treated in addition with high dose cyclophosphamide and Ara-C over 3 weeks. One hundred and one patients received cranial irradiation (1,800 rads standard risk (SR)-patients, 2,400 rads HR-patients) and intrathecal MTX. Maintenance therapy was performed with the usual two drug combination of daily 6 mercaptopurine (6 MP) and weekly MTX orally. Based on phenotyping 67% of patients had common type ALL, and pre-T or T-cell type in 18%. Six per cent of the patients had leukemic blasts expressing both common ALL and T-cell markers (c/T-type); 9% had acute undifferentiated leukemia (AUL). Out of 108, 101 achieved a complete remission, 6 patients died during induction therapy, 1 was a non-responder and 9 patients relapsed. Of these 6 had haematological relapses, 2 had CNS relapse and 1 had a testicular relapse. Four patients died in continuous complete remission (CCR). For 101 patients the 30 months probability of CCR is 0.85 (± 0.05). For 51 patients with standard risk CCR probability is 0.98 (± 0.03), for 50 patients with high risk indices it is 0.65 (±0.11). Patients with c-ALL have a CCR survival of 0.85 (±0.07), those with T- or pre-T-ALL 0.88 (±0.09), all 5 patients with c/T-ALL alive in CCR. In our study pediatric AUL patients have the most unfavourable prognosis.

Acute ITP following infection in children has a high rate of spontaneous remission. Since the platelet count is not uncommonly profoundly depressed, ITP is associated with a mortality of around 1%. The aim of treatment in ITP is to prevent bleeding. As a rule, this can only be achieved by normalizing the platelet count. Intravenous immunoglobulin offers certain advantages over conventional steroid therapy. Moreover, two out of three patients with chronic ITP respond to IgG i.v. The characteristic features of ITP in pregnancy, thrombocytopenia in babies born to mother with ITP and neonatal isoimmune thrombocytopenia are discussed.

The carboxylic esterase (E.C. 3.1.1.1 ) isoenzymes from cases of acute myeloid leukemias were separated by analytical isoelectric focusing on horizontal thin-layer gels. One isoenzyme consisting of one or two components (bands) could be completely and selectively inhibited by addition of 40 mM sodium fluoride ( NaF ) to the staining bath. The 105 cases were classified into the groups M 1 –M 6 according to the FAB proposals. The NaF -sensitive isoenzyme was not detected in cases of FAB groups M12 (acute myeloblastic leukemia without or with maturation), group M 3 (acute promyelocytic leukemia) or group M6 (erythroleukemia). Thirty-one out of 33 cases in the FAB group M 4 (acute myelomonocytic leukemia) and 99 cases in FAB group M 5 (acute monocytic leukemia) expressed the NaF -sensitive isoenzyme. The NaF -sensitive isoenzyme was found at different staining intensities; all M5 cases showed the isoenzyme at strong or very strong intensity, whereas most of the M4 cases displayed the isoenzyme at weak, medium or strong staining intensity. The data presented are further evidence that the presence of the NaF -sensitive esterase isoenzyme indicates monocytic involvement or differentiation in cases of myeloid leukemias. The easy and fast to perform method of isoelectric focusing can be used to distinguish the monocytic variants among the acute myeloid leukemias and can supplement the morphological analysis of these cases.

Receptor binding and biological effects of insulin and insulin-like growth factors I and II (IGF I/II) were assessed in three human malignant cell lines: a Burkitt-type ALL-cell line, a ANLL-cell line and a Hodgkin's disease-cell line. Insulin receptor binding could be demonstrated in Burkitt-type ALL cells and ANLL cells, whereas no insulin receptor binding was detectable in Hodgkin cells. IGF I and IGF II binding could be demonstrated in all leukemic cells. Insulin stimulated glycogen synthesis in the insulin receptor bearing cell lines. DNA synthesis was stimulated by insulin, IGF I and II. IGF I was more active in stimulating DNA synthesis than IGF II.

Fresh leukaemia cells from the peripheral blood of 6 patients with B-chronic lymphocytic leukaemia (CLL) were cultured in the continuous presence of the phorbolester 12-O-tetradecanoylphorbol 13-acetate (TPA) for in vitro induction of differentiation. Upon treatment with TPA the cells showed distinct morphological changes consisting of cytoplasmic and nuclear enlargement, vacuolisation and protrusion of cytoplasm, eccentric location of nuclei with perinuclear clear zones, and oval to elongated cell forms. Isoenzyme profiles of the enzymes carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase (LDH) were analysed by isoelectric focusing on polyacrylamide gels. An increase in the number and in the staining intensity of isoenzymes were observed for all 4 enzymes in the TPA-exposed cells indicating a maturation along the B cell pathway. TPA triggered the new expression of the tartrate-resistant acid phosphatase isoenzyme, a marker of hairy cell leukaemia (HCL) cells, and of the hexosaminidase I isoenzyme, a marker of multiple myeloma cells. The induced phenotypic changes are suggestive of differentiation to stages corresponding to those of HCL cells or 'pre-plasma cells'. Images Figure 2 Figure 4

Two paediatric patients with systemic lupus erythematosus were treated with immunoglobulin G (IgG). In the first case treatment resulted in regression of the most acute symptoms and a long remission was achieved. In the second patient, who was treated during the chronic stage of the disease, there was no significant effect on the course of the SLE.

The phorbolester 12-0-tetradecanoylphorbol 13-acetate (TPA) was used for the induction of differentiation in cells of the human leukemia cell line SPI-802. The cellular morphology, surface marker antigen expression, and isoenzyme profiles of four enzymes (carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase) served as parameters for monitoring the induced phenotypical changes. TPA led to distinct alterations of the morphology and significantly affected the growth rate with cessation of cell proliferation. No major increase in the number of nitro blue tetrazolium-positive cells or aggregation of cells, phagocytosis of latex beads, adherence to plastic surface, or development of pseudopodia were observed. As TPA-treated SPI-802 cells remained negative for these markers of the monocyte-macrophage complex, it can be concluded that the cells did not differentiate into monocytes and macrophages. The immunological marker profile based on testing of 55 monoclonal antibodies, terminal deoxynucleotidyl transferase and two erythrocyte rosette tests indicated a differentiation of SPI-802 cells along the granulocytic cell lineage. This was confirmed by isoenzyme analysis, especially that of carboxylic esterase. An isoenzyme specific for monocytes and macrophages was not detected. In earlier studies it was found that SPI-802 cells produce hemoglobin upon exposure to TPA or hemin. This latter observation and the present results suggested a comparison with the two erythroleukemia cell lines K-562 and HEL. SPI-802 cells appear to have the potential to differentiate along several cell lineages.

In patients with Haemophilia A, the development of inhibitor is a life-threatening complication of treatment. These patients are at high risk for dangerous bleeding as a result of this acquired resistance to human Factor VIII concentrate. Although treatment of bleeding complications has been improved with the introduction of an activated prothrombin complex preparation, therapy remains unsatisfactory. Two patients with Haemophilia A inhibitor were treated with high dose intravenous immunoglobulin in the expectation of an immunosuppressive effect. A rise in the antibody titre at the same time as the administration of factor VIII concentrate showed that this treatment was ineffective in patients with Haemophilia A inhibitor.

In a 14.5 year old girl with Crohn's disease a complete remission was achieved with intravenous immunoglobulins. Earlier treatment with various therapeutical regimens had been unsuccessful. During the next two relapses the same prompt positive response was reproducible, whereas in the third the dose of immunoglobulin had to be doubled.

Thymidine kinase positive (TK + ) N type cell lines that had been transformed by spleen focus-forming virus were established by transformation with NB tropic Friend virus complex. Thymidine kinase deficient (TK - ) cell clones were isolated. Some of these cell clones release 1000- to 100,000-fold reduced amounts of Friend virus complex as compared to the TK + parental cell clone. TK - clones were grown in medium without BrdUrd. Some of these TK - clones can be induced to release endogenous helper virus and transforming spleen focus-forming virus on reexposure to 10 -6 -10 -4 M BrdUrd. The induced Friend virus complex is of N host range as expected with induced endogenous virus in N-type cells. Before the induction of the endogenous virus spleen focus-forming virus complex, an induction of thymidine kinase (ATP:thymidine 5′-phosphotransferase, EC 2.7.1.75) activity is observed. The latter is possibly a prerequisite for the induction of endogenous virus in TK - cells. Induction of thymidine kinase activity and of endogenous virus is transient and always correlated. The role of BrdUrd and another thymidine analogue, azidothymidine, in interfering with C-type virus release in virus positive cells is discussed. Azidothymidine is unable to induce endogenous virus. Induction of endogenous virus by BrdUrd and inhibition of virus release in virus positive cells is apparently not caused by the same mechanism.

The ability of 12-O-tetradecanoylphorbol 13-acetate (TPA) to induce stable phenotypic changes that serve as markers of differentiation was examined in the non-T/non-B leukemia cell lines KM-3 and REH and in the pre-B leukemia cell lines BV-173 and NALM-6. Isoenzymes of the enzymes carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase (LDH), separated by isoelectric focusing on horizontal polyacrylamide thin-layer gels, were used to monitor induced changes. TPA in different concentrations completely or partially inhibited cell proliferation, but had no drug-related cytotoxicity. No increase in the number of nitro-blue-tetrazolium-reducing cells nor adherence to plastic surface was found. In all four cell lines, TPA caused an increase in number and staining intensity of esterase, acid phosphatase, and LDH isoenzymes. The resulting isoenzyme profiles corresponded to those seen at more mature intermediate stages of B-cell proliferation, but did not indicate a terminal differentiation to mature B cells. The loss of the hexosaminidase I isoenzyme, which is a marker of immature hematopoietic cells, was a further indicator of induced maturation. These results demonstrate that while TPA is capable of inducing various immature non-T/non-B and pre-B cell lines to differentiate, the differentiation progression appears to be restricted to intermediate stages, in contrast to the terminal differentiation inducible in myeloid cells.

The clinical application of an antiserum recognizing common ALL associated antigen (cALL-AG) is very useful in classifying leukemias and diagnosing bone marrow relapse as well as CNS-leukemia. We could demonstrate that sera of common ALL (cALL) patients contain cALL-AG; its partial biochemical characterization is described. The anti cALL serum (cALL-AS) was raised in rabbits with cALL-cells precoated with rabbit antiserum against normal human lymphocytes. After appropriate absorbtion the cALL-AS was highly specific for cALL cells. The isolation of serum cALL-AG was performed by ammoniumsulfat precipitation, gel chromatography and affinity chromatography on agarose lens culinaris hemagglutinin A (lentil lectin). The apparent molecular-weight of the serum glycoprotein is 125 000. Two cALL-AG active structures could be solubilized from cALL cell membrane. The apparent molecular-weights were calculated to be 55 000 and 110 000.

Stable human leukemia-lymphoma cell lines provide model systems to study the processes involved in leukemic and normal cell differentiation [10]. Leukemic cells, arrested at a certain stage of differentiation, can be triggered to differentiate to functionally and morphologically more mature cells [7]. Both “fresh” leukemic cells and cells maintained in long-term culture are sensitive to in vitro induction of differentiation. Furthermore, cell differentiation is a novel concept in the treatment of acute leukemias and several substances such as low dose Ara-C and the physiologic Compound retinoic acid have been shown to act as differentiating agents in vivo [7].

The cyclic chemotherapy scheme OS I/75 was tried in 6 patients with newly diagnosed osteosarcoma and in 3 patients with secondary metastases. The treatment consists of high dose methotrexate, followed by citrovorum-factor rescue, doxorubicine (Adriblastin) and cyclophosphamide (Endoxan). All 6 primary patients are in a continuous remission of 6+ to 21+ months (median 12+ months). The length of remission in the patients with metastases is 5.5+ and 8+ months. The haematological side effects led to an average prolongation of the cycle by 11 days in a planned cycle duration of 42 days. However, they were readily manageable. Among the other side effects two cases of Adriblastin myocardiopathy are remarkable which became apparent after methotrexate and ifosfamide. In order to improve possibilities for treatment regional centralisation of patient care and interdisciplinary and supraregional cooperation of treatment centres are necessary. A prospective treatment programme has been developed for the Federal Republic of Germany and Austria.

29 cases of T-cell derived lymphoblastic lymphoma and T-ALL have been analyzed. There is a striking prevalence of the male sex. In the peripheral blood we often find initially an excessive number of white blood cells combined with normal values for the other constituents in about half of the patients; This may be an expression for the rapid occurrence of leukaemia in T-cell lymphosarcoma. In addition to systemic ALL-therapy we performed X-ray irradiation of the mediastinum in 8 of our patients. This yielded to significantly longer first complete remissions. All patients with T-cell LSA/ALL with or without mediastinal mass should be treated in this manner. Cytochemically a strong focal acid phosphatase reaction was found to be acharacteristic of these cells. It has proved to be a screening method for this disease. The cells are T-cell derived and their pattern of surface markers is similar to that found in fetal thymocytes.

Five lymphatic neoplasms with strong focal acid phosphatase reactivity were selected from a group of acute lymphocytic leukemias and lymphoblastic lymphomas. All five cases showed an anterior mediastinal mass and exhibited identical morphology. This type of lymphoma has been described by Lukes under the term "malignant lymphoma of convoluted lymphocytes". Analysis of surface membrane receptors revealed that the tumor cells lacked surface immunoglobulin and receptors for Fc-fragment, but possessed receptors for complement (C3), untreated SRBC (ES) and SRBC treated with neuraminidase (ESN). By applying a mixed rosette assay using nucleated chicken erythrocytes coated with antibodies and C3, and denucleated ESN, it was found that a considerable number of tumor cells in all five cases formed mixed rosettes, i.e. that they bore the C3 receptor characteristic of B cells and simultaneously the E receptor characteristic of T cells. Thus the tumor cells resembled immature thymocytes of 10-15 weeks' gestation, which also show focal acid phosphatase reactivity and simultaneous expression of C3 and E receptors.

The analysis of various enzymes has been found to distinguish immunologic subsets of human leukemias. This is especially true for terminal deoxynucleotidyl transferase (TdT). This enzyme is present in 90% of all cases with common ALL, pre B-ALL, T-ALL, and it is negative in B-ALL. Some 5%–15% of AML are also positive for TdT. Interestingly, acute undifferentiated leukemias (HLA-DR positive, cALL-A negative) have been found to be TdT positive in 40%–60% of cases. This nuclear enzyme thus seems to be a very early marker of lymphohematopoietic development. The lysosomal enzyme s-hexosaminidase (N-acetyl-s-D-glucosaminidase) can be separated into two major forms: an isoenzyme A, which is constituted by two a and two s subunits, and in isoenzyme B, which consists of four s subunits.

Since in a pilot study i.v. IgG was shown to induce a rapid rise of thrombocytes in children with ITP two prospective multicenter ITP studies were started: one comparing i.v. IgG with oral corticosteroids in previously untreated acute ITP, the other investigating the response to i.v. IgG in pretreated acute or chronic ITP in childhood. In this report preliminary results of both studies are summarized. i.v. IgG treatment of acute and chronic ITP is at least as effective as corticosteroid therapy but is not associated with significant side effects. At least some patients with chronic ITP may benefit from i.v. IgG. Longer observation periods are required for further analysis.

Besides the recent advances in immunology that provided important information for the characterization of leukemia cells, enzyme marker analysis is another area of progress in leukemia research. Assays of enzyme activities, both quantitative enzyme levels and qualitative isozyme changes, are of value in the classification of leukemias. The interest in the study of enzyme markers is due not only to technical improvements and new biochemical possibilities, but also to the involvement of enzyme marker analysis in the so-called "multiple marker analysis," which combines traditional and recently developed techniques in leukemia research. The aims of this review are to describe well-known and potential enzyme markers as well as to stress the relevance of enzyme marker analysis combined with multiple marker analysis for leukemia subclassification and the understanding of normal hematopoietic cell differentiation.

ITP in pregnancy may lead to fetal thrombocytopenia caused by the transplacental passage of maternal antiplatelet antibody. The most hazardous complication in the infant is intracranial hemorrhage. In addition ITP in pregnancy is reported to be associated with an increased abortion rate and an elevated fetal morbidity and mortality. Therefore obstetric management must aim at increasing maternal and fetal platelets. Severel therapeutic approaches to the treatment of ITP in pregnancy are evaluated. Two cases of ITP in pregnancy are reported. Administration of high-dose intravenous immunoglobulin is introduced as a new therapy for ITP in pregnancy. The rapid reversal of thrombocytopenia following immunoglobulin G administration suggests that it is useful especially as emergency treatment for ITP in pregnancy.

A case of omento-mesenterial fibromatosis in a 6-year-old boy revealing through acute intestinal obstruction is reported. Mesenterial fibromatosis, also known as aggressive fibromatosis or desmoid tumor is generally associated with Gardner's syndrome but otherwise an extremely rare disease. Fibromatoses do not metastasize but are characterized by a high incidence of local recurrence. Primary treatment is operative. After multiple recurrence irradiation, cytostatics and antiestrogens have been used additionally to control the disease.

Die primaren Targetzellen des HIV sind die T-Helfer (CD4)-Lymphozyten, die an ihrer Oberflache OKT- bzw. CD4- und Leu 3-Antigene tragen (Barre-Sinoussi et al. 1983, McDougal et al. 1986). In zahlreichen Studien konnte nachgewiesen werden, das die Zahl der T4-Zellen nach Serokonversion zeitabhangig abnimmt und als ein wesentlicher Pradiktor fur die Erkrankung von AIDS zu betrachten ist. Im Verlauf der HIV-Infektion werden periphere Zytopenien beschrieben, wobei auser uber Lymphopenien, Anamien und Neutropenien auch uber Thrombozytopenien berichtet wurden (Ubersicht bei Murphy et al. 1987).

Three cases of congenital brain tumours are reported, which presented with signs of increased intracranial pressure soon after birth, giving rise to the suspicion of a cerebral haemorrhage or of hydrocephalus. Correct diagnosis of tumour was established by computerised tomography, which additionally demonstrated a concomitant haematoma in each case. Two of the newborns had a primarily fatal course, with no specific treatment of the tumours being feasible. The remaining child underwent two operations, experiencing an unhindered neurologic and mental development thereafter. The histologic diagnoses were spongioblastoma and medulloblastoma in the first two cases, and ganglioneuroblastoma in the last. A conspicuous clinicopathologic feature of this neuroblastoma was the marked change in its growth pattern, revealing a higher degree of histologic differentiation and less malignant biological behaviour when tumour regrowth occurred. The diagnostic, therapeutic, and prognostic implications for this special clinicopathologic condition are discussed, with a review of the relevant literature.

Sinee 1 December 1979, 149 children with acute lymphoblastic leukemia (ALL) have been treated according to the prospective study protocol ALL 77-02 [5]. Patients’ characteristics are given in Table 1. High risk patients are defined by WBC > 25 X 103/mm3 and/or T cell marker, AUL, or B cell ALL. The treatment protocols for Standard risk and high risk patients are given in Figs. 1 and 2. During consolidation therapy, three intermediate dose methotrexate infusions [2] were administered at 10-day intervals; 24 h after the end of MTX infusion, l-asparaginase and citrovorum factor were given [7]. The second part of consolidating consisted, in therapy group B of high dose cyclophosphamide (X2) and intramuscular Ara-C (X 16) [6].

The isoenzyme profiles of hexosaminidase (N-acetyl-beta-D-glucosaminidase) were analyzed by isoelectric focusing on horizontal polyacrylamide thin-layer gel with special emphasis on the intermediate isoenzyme (Hex I). The expression of Hex I was examined in 87 leukemia-lymphoma cell lines, in 14 B-lymphoblastoid cell lines, in 441 cases of leukemia-lymphoma (specimens containing 80% or more tumor cells), in 22 leukemia cell lines and in 14 cases of leukemia that had been treated with phorbolesters (TPA) for induction of differentiation, and in the mononuclear cell preparations separated from peripheral blood, lymph node, thymus, bone marrow, tonsil, liver, and spleen specimens from normal donors. Hex I was detected in the leukemia cell lines arrested at early, immature or at late, mature stages of B- and T-cell differentiation, but not in cell lines blocked at intermediate stages of maturation. Most myelomonocytic leukemia cell lines and the erythroleukemia cell lines showed Hex I, whereas the B-lymphoblastoid cell lines were negative for this marker. During induction of differentiation, the expression of Hex I was lost in 13 of 15 leukemia cell lines that were originally Hex I-positive. Among the panel of the "fresh" leukemia-lymphoma cells, Hex I was found predominantly in cases of acute lymphoblastic leukemia and acute myeloblastic/monoblastic leukemia, but rarely or not at all in the mature T-, B- or myeloid malignancies. However, two out of two cases of multiple myeloma were Hex I-positive, and the Hex I expression could be induced by TPA in three of six B-cell chronic lymphocytic leukemia cases. Chronic myelocytic leukemia cells remained Hex I-negative during induction of differentiation. Hex I-positivity was not detected in the cell preparations from normal tissues, and peripheral blood indicating that the normal cellular counterpart of the Hex I-positive tumor cells are present at only low percentages within the respective cell populations. It is suggested that Hex I is a marker of early lymphoid and myeloid hematopoiesis that is no longer expressed in intermediate stages of lymphoid differentiation and in later or terminal stages of myeloid differentiation, but that is again detectable in terminally differentiated B-cells. Further studies will focus on identification and isolation of normal Hex I-positive cells.

The cells from 87 leukemia-lymphoma cell lines, 14 B-lymphoblastoid cell lines, 459 cases of leukemia-lymphoma, normal specimens, 22 leukemia-lymphoma cell lines treated with 12-O-tetradecanoylphorbol 13-acetate (TPA) and 14 cases of chronic lymphocytic leukemia (CLL) and chronic myelocytic leukemia (CML) treated with TPA were analyzed for the expression of tartrate-resistant acid phosphatase (TracP) isoenzyme separated by isoelectric focusing. The TracP isoenzyme was seen in the following leukemia-lymphoma cell lines: 4 of 30 T-cell, 2 of 35 B-cell, 1 of 6 non-T/non-B-cell, 1 of 8 myelomonocytic, 3 of 4 erythroleukemia, and 3 of 4 Hodgkin's disease-derived cell lines. The expression of the TracP band could be induced by treatment with TPA in 3 myelomonocytic leukemia cell lines. Among the different types of leukemia-lymphoma cells freshly obtained from patients, the TracP isoenzyme was detected at a high incidence in cases of B-CLL, hairy cell leukemia (HCL), and B-lymphoma. Of the myeloid leukemias, 10% to 20% displayed the TracP isoenzyme. TracP positivity was detected in the peripheral blood, tonsil, bone marrow, spleen, and liver obtained from healthy donors, but not in the thymus. The expression of the TracP band could be newly induced by TPA in cases of CLL and in cases of CML. It is concluded that TracP activity is not specific for HCL, but is found at high incidences in cases of HCL, B-CLL and B-lymphoma. The TracP isoenzyme is not expressed by very immature lymphoid leukemia cells, but by cells arrested at later stages of differentiation of the T- or B-cell lineage, and by some myeloid cells.

The expression of hormonal and growth factor receptors in leukemic cell lines might be heterogeneous due to the admixture of different cell cycle phases and the presence of yet unidentified sublines. Therefore, cell cycle-specific separation of Burkitt type ALL cells was performed by counterflow elutriation, and by limited dilution procedure three sublines of this cell line were obtained. Counterflow elutriation enriched to 60-80% purity for G1-, S-, and G2-phase which was shown by DNA flow cytometry. Insulin and insulin-like growth factor I (IGF-I) binding was investigated in the G1, S, and G2 phase. Insulin binding sites decreased from 10 to 15,000 per cell in G1 to 1,000-5,000 in S and increased to 40-50,000 in G2. The affinity of insulin binding remained constant during the cell cycle. IGF-I binding sites increased from 2,000 per cell in G1 to 5,000 in S and 15,000 in G2. The affinity of IGF-I binding decreased from G1 toward S and then remained constant in G2. The three isolated clonal sublines differed in numbers of insulin and IGF-I binding sites/cell without differences in affinity. The fact that IGF-I shows higher affinity binding during G1 than during S and G2 Burkitt type ALL cells suggests that IGF-I might be important for initiation of proliferation. The reduction in insulin binding sites during S-phase may indicate refractoriness of the cell to insulin during DNA replication.

Monocyte esterase activity was studied in 1,000 doctor-attending patients with normal hematological indices and in 56 patients with non-Hodgkin's lymphoma (NHL) or B chronic lymphocytic leukemia (CLL). The incidence of esterase deficiency was significantly greater in the NHL-CLL patients (7.1%) than in the population group (1.7%; p less than 0.05). In the NHL-CLL group, study of the families showed the esterase deficiency to be a familial characteristic. We postulate that the presence of the anomaly may be either a factor predisposing to the development of the NHL or CLL or a factor indicating a predisposition to these disorders.

Spleen focus-forming virus-transformed erythroleukemic cell clones, which have been established by infection of N type mice with NB trophic Friend virus, continue to release biologically active tfriend virus of NB host range. Dimethylsulfoxide induces erythroid differentiation and a 10- to 100-fold increase in the release of biologically active Friend virus. The increase of Friend virus release is a function of the differentiating erythroleukemic cell. The induced Friend viurs is not the NB tropic Friend virus complex, but shows N host range. The induction of the Friend virus complex is due to simultaneous induction of both spleen focus-forming and endogenous viruses.

In 10 out of 15 children with first relapse of ALL complete remission could be achieved by a combination of Adriamycin + Methylprednisolon followed by Asparaginase + Methylprednisolon. Median remission duration was 24 weeks. The results of this regimen proved superior to other regimens tested.

The initial bone marrow smeasrs of 821 children with ALL have been cytochemically analysed. (PAS-, acid phosphatase-(SPH), peroxydase-(POX), alpha-N-esterase-(EST)reaction) with regard to presenting clinical data and course of disease. Granular reactions of PAS and SPH were not meaningful in this respect. The paranuclear SPH-reaction was associated with high incidence of mediastinal mass, high peripheral blas count and male preponderance. All 5 patients with histories of more than 12 weeks duration belongs to the PAS-type of ALL (cloddy reaction). The PAS-type showed a higher incidence of CNS-leukaemia and generally a less favorable prognosis as compared to the UND-type. Thereby a high proportion of PAS positive cells, independently of other risk factors, indicates a better prognosis. The highterto neglected weak EST-reaction (Grad I and II according to Löffler) seems to signalise a clincally distinct subtype of ALL with a younger age distribution maximum as compared to the UND-type, low incidence of risk factors, high rate of remissions lasting more than 2 years but unfavorable endprognosis.

The expression of a particular alpha-naphthyl acetate esterase isoenzyme which is specific for monocytes was examined in a panel of cultured leukemia-lymphoma cell lines (n = 88), freshly obtained leukemia-lymphoma cells (n = 527), and in fresh (n = 10) and cultured (n = 22) leukemia cells treated with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). The sodium fluoride-sensitive isoenzyme was separated by isoelectric focusing on horizontal thin-layer polyacrylamide gels. The esterase isoenzyme was not detected in untreated or TPA-treated lymphoid, erythroid, or Hodgkin's disease-derived cell lines, but was seen in leukemia cell lines of monocytic origin. TPA induced the new expression of this marker isoenzyme in two leukemia cell lines of promyelocytic and erythroid origin that are known to differentiate along the monocytic-macrophage cell lineage; TPA stimulation increased the staining intensity of the band in monocytoid cell lines. This esterase isoenzyme was found in 92% of the cases classified morphologically as acute myelomonocytic or monocytic leukemia, but only in 3% of the non-monocytic acute myeloid leukemias. All lymphoid or erythroid leukemias or lymphomas were negative. Treatment with TPA of AML and CML cells, which commonly differentiate to monocyte/macrophage-like cells, showed de novo the monocyte-specific isoenzyme. It is concluded that this isoenzyme is a characteristic marker for monocytic leukemia cells and will be a useful tool for the discriminatory identification of the monocytic element in normal and leukemic cells.

The effects of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) and of hemin on the human erythroleukemia cell lines HEL and K-562 were studied using the parameters cellular morphology, cell proliferation, positivity in the nitro-blue tetrazolium (NBT)-reduction test, adherence to surface, development of pseudopodia, and isoenzyme profiles of carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase (LDH) separated by isoelectric focusing on horizontal polyacrylamide gels. TPA and hemin induced similar morphological changes in HEL and K-562, erythrocyte-like cells were seen to be the predominant population after exposure to hemin. Both TPA and hemin led to cessation of cell proliferation and to a dose-dependent decrease in the number of viable nucleated cells. Whereas HEL cells treated with TPA became positive in the NBT-reduction test, adhered to plastic surface and showed pseudopodia, HEL cells exposed to hemin and K-562 cells cultured with TPA or hemin did not show these markers indicative of the macrophage-cell lineage. TPA-treated HEL cells exhibited isoenzyme patterns which were characteristically seen in cells of the monocyte-macrophage complex. 'Myeloid isoenzyme profiles' were found in TPA-treated K-562 cells. As evidenced by the above described phenotypical changes, HEL cells appeared to differentiate along the macrophage and erythroid cell lineages. Induced K-562 cells displayed myeloid and erythrocytic markers.

The human leukemia cell lines HL-60, KG-1, KLM-2, ML-3, THP-1, and U-937 were treated with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). TPA partially or completely inhibited the proliferative activity of the cell cultures. The number of cells with the ability to reduce nitroblue tetrazolium increased in the TPA-treated cell lines HL-60, ML-3, THP-1, and U-937, whereas the cell lines KG-1 and KLM-2 remained nitroblue tetrazolium negative. Except for KG-1 and KLM-2, all TPA-treated cell lines showed varying degrees of strong adherence to plastic surface. The carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase isoenzyme profiles from these cell lines were analyzed by isoelectric focusing on horizontal polyacrylamide gels. The new or stronger expression of an esterase isoenzyme which is specific for monocytes-macrophages was induced in HL-60, ML-3, THP-1, and U-937 but not in KG-1 or KLM-2. The new expression of the tartrate-resistant acid phosphatase isoenzyme was induced in ML-3, THP-1, and U-937. The number of esterase and acid phosphatase isoenzymes and the staining intensity of isoenzymes characteristic for myeloid cells were increased by TPA in all cell lines. The loss of the hexosaminidase I isoenzyme which is a marker of immature hematopoietic cells was noted in KG-1, ML-3, THP-1, and U-937. TPA triggered an increase in number and staining intensity of the lactate dehydrogenase isoenzymes in all cell lines. Some isoenzymatic changes (e.g., monocyte-specific esterase, tartrate-resistant acid phosphatase, hexosaminidase I) appear to correlate with TPA-induced differentiation while other alterations in the isoenzyme patterns do not (e.g., lactate dehydrogenase, other esterase and acid phosphatase isoenzymes). Differentiation of nonmonocytoid cells appears, at the isoenzyme level, to be quite different from that of the monocytoid cell lines.

In a prospective multicenter study 42 thrombocytopenic (less than 30 X 10(9) platelets/l) children with chronic idiopathic thrombocytopenic purpura (ITP) or with acute ITP, dependent on or refractory to corticosteroids, were given 0.4 g i.v. IgG/kg body weight/day on 5 consecutive days and thereafter once a week if the platelet count fell to less than 20 X 10(9)/l or if the patient bled. After the initial 5 days of i.v. IgG the platelets rose within a mean of 7-8 days to greater than 30 X 10(9)/l in all and to greater than 150 X 10(9)/l in 33 of 42 patients (79%). After a mean observation time of 26.6 months 26 of 42 patients (62%) showed a satisfactory long-term effect, i.e. no need for treatment for at least 6 months without bleeding and with no platelet counts below 20 X 10(9)/l. No difference in response rate was found between children with chronic and those with previously treated acute ITP. These results indicate that i.v. IgG could be used to control emergency situations, e.g. to stop bleeding or to prepare a patient for surgery. I.v. IgG also represents a good alternative to treatment modalities, such as splenectomy and/or the administration of cytostatic immunosuppressants with potentially serious side effects. In addition to the expected transient rise in serum IgG levels, i.v. IgG induced a more prolonged elevation of serum IgM. Platelet associated IgG, elevated before therapy, was correlated with the clinical long-term outcome.

Wir berichten uber einen 41/2 Monate alten weiblichen Saugling, bei dem aufgrund der Leitsymptome Hepatomegalie und rezidivierende Hypoglykamien die Verdachtsdiagnose Glykogenose Typ I gestellt wurde. Nach einer Leberbiopsie, die sowohl einen normalen Glykogengehalt der Leber als auch eine normale in vitro Aktivitat der Glucose-6-Phospha-tase ergab, wurde die Diagnose revidiert. Die daraufhin durchgefuhrte Funktionsdiagnostik des Kohlenhydratstoffwechsels ergab aber eindeutig, das in vivo die Umwandlung von Glucose-6-Phosphat in Glucose blockiert ist. Es mus daraus gefolgert werden, das auch bei normaler in vitro Aktivitat der Glucose-6-Phosphatase die Diagnose einer Glykogenose Typ I nicht ausgeschlossen ist. Die Funktionsdiagnostik des Kohlenhydratstoffwechsels gewinnt durch diese Beobachtung an Bedeutung. Als Bezeichnung der Erkrankung schlagen wir den vereinzelt schon gebrauchlichen Begriff Glykogenose Typ I B vor.

The reactivity with monoclonal antibodies (MoAbs) specific for myelomonocytic cells and the expression of a particular esterase isoenzyme were analyzed in 159 cases of acute myeloid leukemias. The incidence of positivity of 16 MoAbs (MCS-2, MCS-1, OKM1, My-1, Leu-M1, Leu-M3, CA-2-38, MY4, MY7, MY8, MY9, VIM-D2, VIM-D5, Mo1, Mo2, 63D3) was studied using the indirect immunofluorescence technique. A carboxylic esterase isoenzyme which can be inhibited completely and selectively by sodium fluoride (NaF) was demonstrated by isoelectric focusing on horizontal polyacrylamide gels. This NaF-sensitive isoenzyme indicated the monocytic origin of the blast cells as it is specific for this cell lineage. Prior to the immunological-isoenzymatic analysis all cases were categorized into two subtypes according to morphological criteria of the FAB classification system: 147 cases of AML (FAB M1-3) and 12 cases of AMMoL/AMoL (FAB M4/5). However, 15 out of 147 cases of AML expressed the NaF-sensitive isoenzyme and were therefore assigned to the group AMMoL/AMoL. Likewise, 1 case, diagnosed morphologically as AMMoL, was negative for this marker isoenzyme and was assigned to the other leukemia subtype. The incidence of reactivity varied widely for the MoAbs tested regarding the overall results on all cases and the positivity on cases of either AML or AMMoL/AMoL. The MoAbs were grouped into four classes depending on the pattern of reactivity with myeloblastic or monoblastic or both subtypes of acute myeloid leukemia. The MoAbs MCS-2, MY7, Leu-M1, and MY9 detected the vast majority of cases with either myelocytic or monocytic involvement (group-I: “pan-myelomonocytic” reactivity). The MoAbs MCS-1, OKM1, VIM-D5, and Mo1 showed a predominance in their staining pattern for monocytic variants, but were also positive on a substantial percentage of nonmonocytic cases (group-II: predominantly reactive with monocytic, but also myelocytic cases). The MoAbs Leu-M3, MY4, VIM-D2, Mo2, and MY8 reacted with the large majority of AMMoL/AMoL cases and with a small number of AML cases (group-III: monocyte-“specific” reactivity). The MoAbs of group-I are useful in differentiating acute lymphoid from acute myeloid leukemias. The MoAbs of group-III, and to a lower extent those of group-II, will be of considerable value in the subtyping of acute myeloid leukemias. The results show that (1) accuracy of leukemia classification might not always be achieved by morphology alone, but that immunological and biochemical aspects should be included as well, and (2) several MoAbs are very useful tools for classification and subtyping of acute myeloid leukemias.

For a number of lysosomal hydrolases, abnormally expressed forms have been found to occur in childhood and adult leukemia. Most of these abnormalities are confined to the physicochemical properties of the enzymes, e.g., abnormal electrophoretic mobility, change of isoelectric point, and alteration of isoenzyme pattern. All this has been described for the hexosaminidase system in human leukemia cells. The most-investigated phenomenon is the occurrence of hexosaminidase I [1–3, 5, 6, 19]. Other abnormalities, such as the anodic shift of the Hex A isoenzyme [2, 6] or a relative decrease of the Hex B form [1], are found in various leukemia subtypes.

B-cell neoplasias such as CLL can be viewed as models of monoclonal populations restricted within discrete ranges of B-cell maturation. It is unknown whether other B-cell leukemias such as prolymphocytic leukemia (PLL), lymphoplasmacytoid immunocytoma (IC), and hairy cell leukemia (HCL) involve different B lineages or are malignant variants of B cells in successive stages of development along the same lineage. Therefore in vitro maturation was induced with the phorbol ester TPA in leukemic cell samples from 10 CLL, 4 PLL, and 4 IC patients. Morphologically, both plasmacytic and hairy cell-like phenomena were induced. The latter unexpected finding was confirmed by reaction with HD6 (CD22) antibody which stains HCL but is unreactive with plasma cells, multiple myeloma, and CLL cells. Tartrate-resistant acid phosphatase was demonstrated in TPA-cultured CLL, PLL, and IC cells, and the same isoenzyme band as in HCL was revealed by isoelectrofocusing. On the other hand, an increase of IgM messenger RNA was detected in up to 20% of the cells in CLL cultures by single-cell in situ hybridization with fluoro-chrome-labeled DNA probes. An abundance of IgM messenger RNA characterizes lymphoplasmacytoid cells as found in IC. Our data demonstrate that CLL, PLL, and IC can be induced to realize a common genetic program which bears characteristics of HCL indicating that these four entities are more closely related than previously thought.

A 12-year old boy with mucoviscidosis developed a mechanical obstruction of the small intestine. The diagnosis of a meconium ileus equivalent was made and a conservative treatment was started. After this treatment failed, a laparotomy was performed which verified the diagnosis of a meconium ileus equivalent. It was possible to move the inspissated feces inside the terminal ileum into the ascending colon. The postoperative course was uneventful. The etiology, differential diagnosis, and both conservative and operative treatment of this complication of the mucoviscidosis are discussed.