Proliferation of human progenitor cells in a long-term culture system is more efficiently sustained by the addition of Flt-3 ligand or megakaryocyte growth and development factor than by Kit ligand.

INTRODUCTION: We compared the effects of the early-acting growth factors (GF), Flt-3 ligand (FL), c-Kit ligand (KL), and leukemia inhibitory factor (LIF), and the late-acting GF, granulocyte-colony stimulating factor (G-CSF) and megakaryocyte growth and development factor (MGDF), added alone in human long-term marrow culture (LTMC). MATERIALS AND METHODS: The GF were used in primary cultures of mononuclear cells (MNC) and in cocultures of CD34+ cells on murine preestablished MS-5 stromal layers. GF activity was assessed as nonadherent and adherent progenitor cell production and cobblestone area formation at week 5. RESULTS: In this system, only FL, KL, and MGDF significantly stimulated early stages of hematopoiesis, whereas only G-CSF stimulated the proliferation of mature progenitor cells within the granulo-monocyte lineage and no effect was observed with LIF. FL displayed the strongest activity, and MGDF was more efficient than KL, both in primary cultures of MNC and in cocultures of CD34+ cells. However, the stimulatory effects of these GF used alone were dependent on the presence of a stromal layer. CONCLUSION: These LTMC data emphasize the particular roles for FL and MGDF in the stimulation of primitive hematopoiesis.

Persistent decrease in proliferative potential of marrow CD34(+)cells exposed to early-acting growth factors after autologous bone marrow transplantation.

Post-graft hematopoiesis is characterized by long-term quantitative deficiency in marrow progenitor cells in both autologous and allogenic settings. In order to evaluate the function of post-graft progenitor cells, the proliferative capacity of marrow CD34(+) cells was evaluated in 10 patients 6 months after autologous bone marrow transplantation (ABMT) for non-Hodgkin's lymphoma and compared to that of 10 patients before ABMT and 10 normal controls. Immuno-selected CD34(+) cells were cultured for 7 days in liquid serum-free medium with a combination of early-acting GF consisting of stem cell factor, IL-3 and IL-1beta. Clonogenic efficiency of unselected cells for CFU-GM and BFU-E was decreased in post-graft patients compared to pre-graft and control patients. However, clonogenic efficiency of selected CD34(+) cells for CFU-GM was not different in post-graft, pre-graft and control patients but BFU-E values of post-graft patients remained lower than those of control patients. Decreased percentages of CD34(+) CD38(-) cells were observed in both post-graft and pre-graft patients while those of CD34(+) c-kit(+) cells were similar in all three patient groups. After 7-day liquid culture, expansion yields of total progenitor cells were significantly lower in post-graft patients (147 +/- 28%) than in pre-graft (255 +/- 27%) and control patients (246 +/- 23%). Post-graft deficiency in progenitor cell expansion was particularly marked for BFU-E (61 +/- 24%) compared to pre-graft patients (220 +/- 82%) and to controls (349 +/- 82%). These results indicate impaired proliferative potential of marrow CD34(+) cells several months after ABMT involving erythroid progenitor cells and/or commitment towards erythroid lineage from a more immature stage (pre-CFU).

Role of early splenectomy in malignant lymphomas with prominent splenic involvement (primary lymphomas of the spleen). A study of 59 cases.

The outcomes were analyzed retrospectively of 59 cases of non-Hodgkin lymphoma (NHL) that included prominent splenic involvement (LPS). Forty-three patients had low-grade NHL, and 16 had intermediate or high-grade NHL. Forty of the 59 patients underwent splenectomy. Four patients died postoperatively before any treatment, and 10 others received no chemotherapy or radiation therapy. Twenty-nine splenectomized and 16 patients whose spleens were not removed received chemotherapy or radiation therapy. One or more cytopenias were present in 45 patients (77%). Five (18%) of the 28 patients who initially were cytopenic underwent splenectomies that did not correct their blood disorders. The median actuarial survival was 108 months in splenectomized patients and 24 months in those not treated surgically (P = 0.0001). For the 40 splenectomized patients, a normal postoperative platelet count, an initial hemoglobin level of 110 g/l or more, and a postoperative hemoglobin level 110 g/l or more were associated with prolonged survival. These results suggest that cytopenias are frequent in LPS and that their reversal is observed after early splenectomy in 82% of cases. The absence of cytopenia after early splenectomy is associated with prolonged survival.

Acute monocytic leukaemia in adults: treatment and prognosis in 99 cases.

Acute monocytic leukaemia (AMoL) was diagnosed in 99 adults, aged 18-85 years (median 56) over a period of 10 years. Sixty-five patients had extramedullary leukaemia, 13 had clinical signs of leucostasis, and 19 had disseminated intravascular coagulation. Four patients died before receiving any treatment, 12 received supportive care only and seven received low dose AraC, but only one of them responded. Seventy-six patients received intensive chemotherapy, 72 of them with an anthracycline-AraC based regimen, with or without an epipodophyllotoxin. Fifteen patients died within 7 d of diagnosis, due to leucostasis in nine cases. Predictive factors for early death were advanced age, leucostasis, fever, leucocytes above 100 x 10(9)/l, and renal failure. Fifty (66%) of the patients treated intensively reached complete remission (CR). Advanced age, fever and complex cytogenetic abnormalities were significantly associated with a lower CR rate. Median actuarial disease-free survival was 20.5 months, and was not significantly influenced by any pretreatment parameter. Five patients relapsed in the central nervous system (CNS), in spite of systematic CNS prophylaxis. No differences in CR rates were seen with the three anthracycline-AraC based regimens used in our patients. Significant differences in disease-free survival were seen between them, however, suggesting that early consolidation chemotherapy and, more hypothetically, epipodophyllotoxin agents could prolong remission duration in AMoL.

Prognostic value of dysmyelopoietic features in de novo acute myeloid leukaemia: a report on 132 patients.

The prognostic value of cytological features was assessed in 132 patients with de novo acute myeloid leukaemia (AML) treated by anthracycline-cytosine-arabinoside combination chemotherapy. Of these patients, 98 (75%) achieved complete remission (CR). A significantly lower CR rate was seen in patients with trilineage dysmyelopoiesis (TDMP) (P = 0.003), but not in patients with dyserythropoiesis and/or dysgranulopoiesis without abnormal megakaryocytes. Other unfavourable factors were age greater than 50 years (P = 0.042), leucocyte count greater than 100 x 10(9)/l (P = 0.006), M5 FAB subtype (P = 0.013), presence of complex cytogenetic rearrangement or abnormalities of chromosome 5 and/or 7 (P = 0.001). Bone marrow eosinophilia greater than 3% was significantly associated with a higher CR rate (P = 0.04). In a multivariate analysis, a low CR rate was best predicted by the presence of a complex karyotype or abnormalities of chromosome 5 and/or 7 (P = 0.0001) and by the TDMP (P = 0.0036). Median actuarial disease-free survival (DFS) was 24 months. Actuarial DFS was significantly shorter in patients with TDMP (P = 0.0001) and an elevated leucocyte count (P = 0.02). Age, FAB subtype and karyotype had no significant incidence on DFS. Presence of TDMP appears to be an important prognostic factor in de novo AML. This could be used as one of the guidelines to therapy.

[Acquired idiopathic sideroblastic anemia. Apropos of 39 cases]. / Les anémies sidéroblastiques acquises idiopathiques. A propos de 39 cas.

We report a series of 39 cases of acquired idiopathic sideroblastic anemia accounting for 12 p. 100 of the 330 cases of myelodysplastic syndrome diagnosed in our department in six years. Patients' mean age was high (70.8 years). Anemia was present in 92 p. 100 of the cases and was of the macrocytic type in 87 p. 100 of the patients. Only two patients (5 p. 100) had neutropenia and two (5 p. 100) had thrombocytopenia. Thrombocytosis was observed in 17 cases (43 p. 100); it was usually moderate. The mean proportion of medullary ring-sideroblasts was 35 +/- 14 p. 100. A clonal abnormality was found in 4 of the 11 karyotype examinations performed. The course of the disease was marked by recurrences of anemia which required regular blood transfusions in 82 p. 100 of the cases. In at least 30 p. 100 of the patients followed up for more than 30 months, these transfusions were responsible for secondary haemochromatosis. Conversely, in only three patients (7.5 p. 100) the disease progressed to acute leukemia. On the other hand, 4 patients (10 p. 100) developed thrombocythemia (greater than 1,000 x 10(9)/1). Median actuarial survival rate was 48 months. There were only two factors of poor prognosis: age over 70 and anemia with less than 8 g/dl haemoglobin at the time of diagnosis. Transfusion haemochromatosis is the principal risk in acquired idiopathic sideroblastic anemia: this risk can be prevented by iron chelating agents in patients with frequent blood perfusions.

Translocation t(10;17)(p13;q12) in two cases of acute nonlymphocytic leukemia with phagocytic activity of blasts. A new cytogenetic entity?

We report two cases of translocation t(10;17)(p13;q12) found in a series of 278 cytogenetically studied acute nonlymphocytic leukemia cases. Blast cells, in both cases, were undifferentiated and had phagocytic properties. These patients might represent cases of a new cytogenetic entity.

Translocation t(1;19)(q23;p13) in acute lymphoblastic leukemia. A report on six new cases and an unusual t(17;19)(q11;q13), with special reference to prognostic factors.

We report clinical, immunologic, and cytogenetic characteristics of six patients with a t(1;19)(q23;p13) that was balanced in one case and of the unbalanced type [-19,der(19)t(1;19)(q23;p13)] in the remaining five cases. Intracytoplasmic immunoglobulins (cIg) were positive in the three cases where they were found. We also report on another patient, with a t(17;19) involving 17q11 and probably 19q13 regions, although involvement of 19p13 could not be excluded. In this patient, cIg were also present, thus raising the issue of whether such a rearrangement could be a variant of t(1;19). Clinically, five patients belonged to the high-risk acute lymphoblastic leukemia (ALL) group, because of high leukocytosis, central nervous system (CNS) disease at presentation, or massive organomegaly. Cytologically, all cases were FAB type L1. Except for the two cases allografted in the first complete remission (CR) all patients relapsed, three of them within 13 months. Two CNS relapses were seen in spite of adequate CNS prophylaxis. ALL with t(1;19) appears to be a poor-risk ALL subgroup and probably requires a reinforcement of therapeutic modalities that might include, when possible, allografting at first CR.

Acute lymphocytic leukemia with 9p anomalies. A report of four additional cases and review of the literature.

Childhood acute lymphocytic leukemia (ALL) with partial deletion of the short arm of chromosome 9 (9p-), particularly in the p21-22 region, associated with bulky disease, has been regarded as a possible subgroup of ALL. We have reviewed clinical and cytologic data in 128 cases of ALL (childhood and adult). Four of them had 9p anomalies. Two patients had a deletion in the 9p21 region associated with another deletion (9p13----pter) in one case and with t(1;19)(q21;p13) in the second patient. A third patient had a t(9;14)(p21;q12) balanced translocation associated with 14q22----qter deletion; the last patient showed a t(5;9)(p14;q21) unbalanced translocation also associated with 14q deletion. All four patients had lymphomatous ALL, but immunophenotype was non-T, in the four cases, (non-T, non-B in two patients and common ALL in the two remaining cases). Acute lymphocytic leukemia with 9p anomalies appears relatively frequently and is usually associated with poor prognostic features (i.e., bulk disease and high leukocyte counts) but does not seem restricted to childhood and T-cell lineage.

Refractory anaemia according to the FAB classification: a report on 69 cases.

Between 1980 and 1986, we diagnosed refractory anaemia (RA), according to the FAB classification, in 69 patients, who constituted 22% of the 312 cases of myelodysplastic syndromes (MDS) seen over that period. The haematological features were variable, with pancytopenia in 14 cases (20%), bicytopenia in 24 (36%) and mono-cytopenia in the remaining patients, including 21 (30%) cases of anemia alone, 8 (12%) cases of refractory neutropenia and 2 (3%) cases of refractory thrombocytopenia. Myelodysplastic features were also quite variable, involving one, two or all three lineages. In patients with a single cytopenia or only one dysplastic lineage, FAB criteria appeared insufficient for adequate inclusion among RA and we suggest more precise diagnostic criteria, resulting from the utilization of cytogenetics, ferrokinetics, progenitor cultures and perhaps molecular biology, in such cases. Median survival was 42 months. 12 patients (17%) progressed to RAEB (of whom 7 finally developed ANLL) and 4 patients (6%) to CMML. In spite of the heterogeneity of haematological features, only two factors were associated with poor prognosis, namely age greater than 70 yr at diagnosis and haemoglobin less than 10 g/dl, whereas, to a lesser extent, neutropenia was associated with progression to RAEB.

Promyelocytic blast crisis of Philadelphia-positive thrombocythemia with translocations (9;22) and (15;17).

We report a promyelocytic blast crisis in a case of Ph-positive thrombocythemia with both t(9;22) and t(15;17). Our patient confirms the specificity of t(15;17) in malignant proliferation of promyelocytes and suggests its appearance as a second genetic event in the genesis of blast crisis occurring in a Ph-positive clone.

Total peroxidase deficiency in eosinophils: a report on twin sisters, one with a refractory anaemia.

A total lack of EPO was fortuitously discovered in a 63-yr-old woman from the north of France who had primary refractory anaemia, but without dysgranulopoiesis; MPO activity was normal. Her twin sister, whose blood count was normal, also had EPO deficiency. This familial disorder was first described in the Israeli Jewish population and is very rare in Caucasians; it seems to have no pathological consequences. Microscopic studies showed no anomaly except a negativity of Sudan Black B staining which we consider to be a sign of peroxidase deficiency. Ultrastructural studies of the granules revealed normal cristalloid, but the matrix, which contains EPO in normal eosinophils, was very thin; the nature of the relation between functional and morphological anomalies has still to be elucidated.

[Cytological diagnosis of myelodysplastic syndromes. Apropos of 200 cases]. / Diagnostic cytologique des syndromes myélodysplasiques. A propos de 200 observations.

The authors present a cytological study of 200 cases of myelodysplastic syndromes (MDS) [corrected], classified according to the FAB cooperative group. This analysis involves the conventional parameters of the peripheral blood and bone marrow differential counts, the systematic recording of signs of dysmyelopoiesis on the erythroblastic [corrected], granulocytic and megacaryocytic lineages and the assessment of blood granulocytes myeloperoxidase (MPO). On the outside of the typical acquired idiopathic sideroblastic anaemia (severe, isolated anaemia associated with intense dyserythropoiesis), the diagnosis of MDS requires very astute cytological interpretation: indeed, when the dysgranulopoiesis is pronounced, determination of peripheral blood and bone marrow differential count is especially difficult. Conversely when the dysmyelopoiesis is slight, a systematic search of morphology signs [corrected] of its presence must be done and associated with functional studies such as MPO activity technically easy to realize.

[Acute lymphoblastic leukemia, Burkitt-cell type, after Hodgkin's disease. Study of 2 cases]. / Leucémie aiguë lymphoblastique à cellules de Burkitt après maladie de Hodgkin. Etude de deux observations.

Two cases of Burkitt cell leukaemia (ALL3) occurring 60 and 51 months respectively after treatment of Hodgkin's disease (HD) are described. Clinical, cytological, immunological and cytogenetic features in these patients were comparable with those of the 6 previously published cases and with de novo ALL3. The prognosis of these secondary ALL3 is uniformly poor: our 2 patients died 2 and 14 days after the start of combination chemotherapy, and all the other cases survived less than 60 days after diagnosis. The significance of ALL3 occurring after HD is discussed: the association is probably not fortuitous, and its pathogenic mechanisms probably differ from those intervening in secondary myeloid leukaemias.