Prognostic value of immunological markers in acute myeloblastic leukemia.

In order to investigate the prognostic value of immunologic markers together with the most relevant clinical and hematological disease characteristics in acute myeloblastic leukemia (AML), the reactivity of blast cells from 102 patients with AML was analyzed with a panel of twenty monoclonal antibodies. The univariate analysis showed that five parameters had an adverse effect on both complete remission (CR) and survival: advanced age (greater than 60 years), anemia (hemoglobin concentration (Hb) less than 10g/dl), the expression of the antigens detected by the anti-megakaryocytic antibodies (CDw41/CDw42), the monocytic antibodies (CD14), and the CD9 (FMC56, FMC8) antigen. In addition, the failure to obtain CR had a significant adverse effect on survival (p less than 0.0001). Multivariate analysis showed that only age and Hb had a significant influence on CR while for predicting survival the most important independent prognostic factors were: CR, age, number of platelets and reactivity with the CD9 antibody. These results show that immunological markers could represent a valuable tool in the assignment of risk categories in AML patients.

alpha Interferon in the management of essential thrombocythaemia.

Thirteen patients (mean age 60.7 years; female:male ratio 10:3) with essential thrombocythaemia were treated with 3 million units (MU)/day interferon alfa-2b subcutaneously (s.c.) for 12 weeks, with all patients requiring a dose reduction after 4 weeks. The mean pretreatment platelet count was 1,400 x 10(9)/L and megakaryocytes were increased in all cases. Splenomegaly was present in six patients and haemorrhagic phenomena were observed in two. Nine patients (69.2%) had objective responses, including two (15.4%) complete responses (platelets less than 450 x 10(9)/L) which were then maintained with 5 MU interferon twice a week. Acute toxicity consisted of flu-like symptoms in 12 patients. Chronic toxicity (mainly leucopenia) was observed in nine patients. In conclusion, initial therapy and then requiring maintenance therapy at a reduced dose. However, the frequent side effects observed make it advisable to use a low dose of interferon alfa-2b, and to treat only those patients with significant symptoms and signs of thrombocytosis.

High levels of plasma FVIII and vWF in the toxic epidemic syndrome patients.

Factor VIII and von Willebrand factor proteins were evaluated in 115 patients having the chronic phase of the Toxic Epidemic Syndrome (TES), a new multisystemic disease probably caused by the ingestion of denatured rapeseed oil, and in 50 control volunteers. Higher circulating levels of factor VIII procoagulant activity (VIII:C) (158 +/- 58.4 U/dl), von Willebrand factor antigen (vWF:Ag) (166.1 +/- 55.5 U/dl) and von Willebrand factor ristocetin cofactor activity (vWF:RCo) (178.7 +/- 55.2 U/dl) were seen in TES patients (p less than 0.001, TES patients versus control subjects, for each parameter). The increased levels of vWF:Ag and vWF:RCo observed in TES patients correlated with the scleroderma like lesion of the skin, with the sicca syndrome and with Raynaud's phenomenon (p less than 0.01), but not with other clinical manifestations. The multimeric analysis of vWF in 92% of the TES patients was similar to that found in normal plasma, but in the remaining 8% a very slight increase of larger vWF multimers in plasma were observed. The raised levels of vWF found in TES patients in the chronic phase may reflect an "in vivo" vascular injury.

Effect of dextran on factor VIII/von Willebrand factor structure and function.

Factor VIII/von Willebrand factor was analyzed before and after the infusion of 500 ml of Dextran 70 to normal volunteers. Factor VIII procoagulant activity, factor VIII related antigen and ristocetin cofactor activity showed a significant decrease, reaching after six hours the minimum level, which did not correlate with the hemodilution effect caused by dextran. Ristocetin-induced platelet agglutination (RIPA) in volunteers' platelet-rich plasma (PRP) did not show any significant change between preinfusion time and six hours after the infusion. Multimeric analysis of von Willebrand factor (vWF) showed a progressive decrease of all the multimers which was more pronounced in the largest multimers. No change was seen in the "triplet" structure of vWF. No effect was noticed when dextran was incubated "in vitro" either with PRP or platelet-poor plasma. The modification induced by dextran is close to the pattern seen in subtype Ib von Willebrand's disease.

Congenital deficiency of vitamin K-dependent coagulation factors and protein C.

A 15-month-old girl from Coimbra (Portugal) had a history of numerous hemorrhagic episodes with multiple bruises, hematomas but not hemarthroses. On serial testing she showed deficiency of factors II, VII, IX, X and protein C. Malabsorption-induced vitamin K deficiency, liver disease or ingestion of a coumarin compound were excluded. An absence of detectable abnormalities was found among her relatives. Consanguinity was not present. The immunologic assay, immunoelectrophoresis or antibody neutralization, revealed much higher levels of these factors than the clotting assay. The non-physiological activator (Echis carinatus venom) produced higher levels of prothrombin activation than those detected by physiological activation. Two-dimensional immunoelectrophoresis of the patient's plasma in calcium showed that prothrombin had the same mobility as acarboxyprothrombin. No significant response to large doses of intravenous vitamin K3 (6 mg) was observed. Transfusion of 120 ml of frozen fresh plasma led to an immediate increase in the procoagulant activities of vitamin K dependent protein, similar to that found after perfusion of plasma plus vitamin K3. The results obtained from this patient suggest a defect in the gammacarboxylation mechanism inside the hepatocyte.

Discrepancies between morphologic, cytochemical, and immunologic characteristics in acute myeloblastic leukemia.

This study was designed to compare the cytochemical pattern with the immunologic phenotype in 108 cases of acute myeloblastic leukemia (AML) classified according to the French-American-British (FAB) criteria. Special attention was paid to the cases where discrepancy existed between these approaches and to a group of 11 patients considered as unclassifiable mainly because a second cell population--megakaryoblastic--was detected. Three types of discrepancies were observed: cases with typical morphologic characteristics and cytochemistry but devoid of lineage-specific antigens; these mainly include poorly differentiated leukemias (eight M1, four M2, and eight M5a), suggesting that the cytochemical enzymes are earlier myeloid markers than the currently available monoclonal antibodies; cases in which immunologic characteristics were discordant with morphologic characteristics and cytochemistry; these include two M2 cases positive for monocytic monoclonal antibodies (CD14); six M5b cases positive for granulocytic monoclonal antibodies (CD15); and seven M4 cases lacking in CD14 or CD15 antigens; cases with discrepancies between morphologic characteristics and cytochemistry and in which the immunologic markers permitted the correct assessment of cell lineage (six cases). These results show that the classification of these patients is better achieved by a combined morphologic, cytochemical, and immunologic approach.

Heterogeneity of T cell lymphoblastic leukaemias.

Twenty eight out of 170 consecutive cases of acute lymphoblastic leukaemia (ALL) were examined. They were of T cell origin, with the following distribution: seven (28%) cases had pre-T or prothymic features; nine (36%) cases showed early thymocytic features, six (24%) had cortical features; and three (12%) had a "mature" phenotype. The remaining three cases could not be sub-classified. A striking finding was that pre-T ALL differed from intrathymic ALL not only in the absence of both E rosettes and intrathymic differentiation antigens, but also in the expression of two non-lineage specific antigens HLA-DR and CD10. Both antigens appear in the bone marrow from the very first stages of lymphoid differentiation, implying that the origin for pre-T ALL is bone marrow. A comparison of the clinical features of pre-T and thymic ALL showed that pre-T ALL disease showed a pattern more similar to non-T ALL disease: a lower incidence of mediastinal mass, absence of extrahaematopoietic disease, lower white cell counts and haemoglobin concentrations, and a higher incidence of bone pain. No obvious difference in response to treatment was apparent. The results show that T-ALL is not only a heterogeneous immunological group but also suggest that it may have different origins: bone marrow for pre-T ALL and the thymus for thymic ALL.

Development of acute leukaemia after idiopathic myelofibrosis.

AIMS: To determine the characteristics of blastic transformation of idiopathic myelofibrosis. METHODS: The clinical and haematological features, as well as the morphological characteristics of blast cells, were analysed in nine adults with blast transformation. RESULTS: Most of the patients were male and had enlarged spleens and livers. Five of the patients had normal platelet counts, while all had pronounced anaemia and a moderate degree of leucocytosis. The duration of the acute phase was usually short: 16 (SD 8) weeks. Most myeloid cell lineages--granulocytic, monocytic, and megakaryocytic--were similarly distributed. One patient also had a hybrid (lymphoid-myeloid) phenotype. The morphological assessment of blast cells agreed with immunophenotyping in five out of the nine cases. The onset of the blastic phase was not related to previous treatment. CONCLUSIONS: A pluripotential stem cell with preferential myeloid commitment would be the target cell of blast transformation in idiopathic myelofibrosis. Our immunophenotypic data do not support the concept of a preferential association between megakaryocytic lineage and the acute transformation of idiopathic myelofibrosis. The absence of previous treatment in some cases suggests that this kind of evolution is part of the natural history of idiopathic myelofibrosis.

Acute phase reactant proteins in differential diagnosis of monoclonal gammopathy.

The serum levels of five individual serum proteins, i.e. ceruloplasmin, alpha 1-antitrypsin, orosomucoid, haptoglobin and transferrin, were studied in 55 multiple myeloma (MM) patients, in 34 essential monoclonal gammopathy (EMG) patients, in 14 EMG patients excluded for active inflammatory process and in 14 healthy control subjects. Transferrin--negative acute phase reactant (APR)--was significantly decreased and the remaining proteins under study--positive APR--slightly increased in the EMG group compared with healthy controls. Transferrin and ceruloplasmin levels were significantly different when the MM was compared to the EMG group so that these parameters might be useful in differential diagnosis between both groups. In the MM clinical stage III, the differences are even more significant. In the IgG3 MM the APR levels seem to be more significantly changed than in the IgG1 and IgG2 MM. In EMG patients with active inflammatory process who were excluded ceruloplasmin, alpha 1-antitrypsin, orosomucoid and haptoglobin were significantly increased when compared with the EMG group.

Acute phase reactants and clinical stages in multiple myeloma.

Serum levels of seven specific proteins mostly acute phase reactants (APR) have been studied (transferrin, alpha 2-macroglobulin, alpha 1-antitrypsin, haptoglobin, C3, ceruloplasmin, orosomucoid) in 14 healthy subjects and in 55 patients with multiple myeloma. The alpha 2-macroglobulin and transferrin are significantly decreased in the myeloma group compared with healthy controls whereas the remaining proteins under study are elevated, significantly only orosomucoid and ceruloplasmin. The APR in the IgG3 myelomas seem to show differences from those of the IgG1 and IgG2 subclasses the levels of the negatively changed proteins being lower, whereas the positive APR higher in the IgG3 myeloma. In the IgA myeloma, however, decreased levels of haptoglobin and alpha 1-antitrypsin have been found in spite of being positive APR. Transferrin and haptoglobin serum level can be included as a new parameter in regression equations for calculation of IgA myeloma cell mass.

Immunological markers of peripheral blood and lymph node lymphocytes in Hodgkin's disease.

The lymphoid subpopulations of peripheral blood and cell suspensions of lymph nodes were investigated in a series of 20 patients with Hodgkin's disease (HD) by conventional surface markers and a panel of 11 monoclonal antibodies (McAb). In peripheral blood of HD, the number of T lymphocytes and the distribution of helper and suppressor/cytotoxic subpopulations was normal, suggesting that the alteration in cell immunity in this condition is either due to a functional defect in T cells or to an alteration in the immunoregulatory mechanisms which are not directly dependent on T lymphocytes. The lymph nodes involved by HD showed an increase in the number of T helper lymphocytes (OKT4+) as compared with the reactive lymph nodes from 14 subjects used as a control group. In involved lymph nodes there was a good correlation between the number of lymphocytes reacting with the McAb OKT3 and EEAT rosette--forming cells, whereas in most hyperplastic nodes the number of T3 lymphocytes was greater than that of E+ rosettes. Hodgkin and Reed-Sternberg cells did not react either with T, B lineage specific McAb or with myelomonocytic (OKM1) or monocytic (Mo2, FMC-17, FMC-33) McAb. These results would make a monocyte-macrophage lineage origin unlikely also for these cells.

[Post-transfusion graft versus host disease in a patient with Hodgkin's disease]. / Enfermedad del injerto contra el huésped postransfusional en una paciente con enfermedad de Hodgkin.

A female with Hodgkin's disease developed graft versus host disease (GVHD) after the administration of two units of packed red cells. Ten days after the transfusion she developed fever and rash, with subsequent hepatic and intestinal disease and a profound bone marrow aplasia. She died from a Candida tropicalis sepsis. The diagnosis of GVHD was made on the basis of clinical and histological criteria. We review this uncommon complication of hemotherapy, with special emphasis on its differential clinical features and its prevention.

[Renal disease in myeloma. Role of the tumor cell mass (author's transl)]. / Nefropatía del mieloma. Papel de la masa tumoral.

Out of a group of 57 patients with the diagnosis of multiple myeloma fourteen (25%) with different degrees of renal disease were selected. Bence-Jones (BJ) proteinuria, infections, and, above all, the tumoral cell mass were the three main factors implicated in the development of myeloma associated renal disease. Only 13% of IgG myelomas presented with renal failure as compared to 27% of IgA myelomas. The patients with BJ and IgD myeloma, classically those with a higher tendency to develop renal disease (in our series 37% of BJ myelomas and 50% of IgD myelomas had renal disease), had the biggest tumoral cell mass of all patients studied. The relationship between tumor cell mass and renal disease in myeloma is supported by the recovery of renal function in a patient with chronic renal failure after a treatment-induced reduction of the tumoral cell mass from 1.71 to 0.82 x 10(12) cells/m2 body surface.

[Antibodies against hepatitis C virus in hemophiliacs: correlation with peripheral blood lymphocyte populations]. / Anticuerpos frente al virus de la hepatitis C en hemofílicos: correlación con las poblaciones linfocitarias de sangre periférica.

The presence of antibodies (Abs) against hepatitis C virus (HCV) was analyzed in 26 haemophiliac patients; a prevalence of 38% was found in the total series and 56% in the treated patients. There were no cases with a positive serology among the three patients who had only received pasteurized factor. The frequency of detection of anti-HCV was higher in the patients who had a positive serology for HIV (67%) and hepatitis B virus (56%) than in the seronegative patients (24 and 29%, respectively). The patients with HCV Abs showed a decrease in the helper (CD4+) lymphocytic population, mainly due to the decrease in the helper inducer (CD4+/Leu8-) cells, such alterations being more evident in the patients who also were HIV+ who showed as well an increase in the T-cell activated lymphocytes (CD3+/la+) and a decrease in CD16+ natural killer cells. These results suggest that the lymphocytic alterations found in the patients with anti-HCV Abs are not specific and would rather be related to the presence or not of HIV infection.

[Clinico-hematological characteristics of acute transformation of chronic myeloproliferative syndromes]. / Características clinicohematológicas de la transformación aguda de los síndromes mieloproliferativos crónicos.

BACKGROUND: The aim of the present study was to analyze the blastic transformation occurring during chronic myeloproliferative diseases (CMPD) and to establish the differences between them. METHODS: The clinical and hematological characteristics of 54 patients in blastic crisis (BC) of a CMPD were analyzed: 40 chronic myelogenous leukemias (CML), 9 idiopathic myelofibroses (IMF), 4 polycythemia vera (PV), and one essential thrombocythemia (ET). The results were analyzed by the BMDP statistical program. RESULTS: The BC of CML appeared in younger patients (p less than 0.05). Only in this group did some patients achieve complete remission. Moreover, in this BC a greater incidence of visceromegalies and leukocytoses were observed. The BC of the IMF patients led to marked anemia (p less than 0.01) and bone marrow infiltration (p less than 0.05); these leukemias were more frequent in males and began with lymphadenopathies and visceromegalies. The transformations of PV were preceded by a longer chronic phase and had a lower incidence of visceromegalies and a nearly normal hemoglobin count value. The patient with acute leukemia secondary to ET did not display visceromegalies but did have anemia, leukopenia and a normal platelet count. None of the four groups responded to therapy, and their survival was short. CONCLUSIONS: Blast transformations of different myeloproliferative disorders have their own idiosynchratic clinical and hematological characteristics, some of these may be related to the chronic phase of disease.