Multivariate analysis of prognostic factors in acute myeloid leukemia: value of clonogenic leukemic cell properties.

The initial clinical and biological parameters, including clonogenic leukemic cell (CFU-L) assay, were reviewed for their prognostic significance in a cohort of 188 adult patients with newly diagnosed untreated acute myeloid leukemia (AML). Almost all patients received induction therapy with daunorubicin (DNR) and cytarabine (Ara-C) according to the European Organization for Research and Treatment of Cancer (EORTC) AML 5 to AML 9 trials. Bone marrow samples from 116 representative patients were obtained for CFU-L assay with an efficiency percentage of 89.6%; 76 patients had a measurement of the CFU-L self-renewal capacity (second plating efficiency [PE2]) and 91 patients had CFU-L inhibition test after exposure to DNR and/or Ara-C. The prognostic significance of parameters such as age, hematological antecedent, WBC count, liver enlargement, and Auer rods is confirmed in the present study. Moreover, high platelet and polymorphonuclear counts appeared to be related to resistance to induction course. However, through multivariate analysis, CFU-L sensitivity to drugs and self-renewal capacity appeared to be major independent prognostic factors in AML. A low CFU-L inhibition in the presence of the DNR and Ara-C combination correlates with a poorer complete remission (CR) rate, but not with CR duration. Patients with the lower PE2 values experienced both higher CR rate and longer CR duration. The practical interest of CFU-L study remains to be defined but, at least, PE2 measurement could be considered in the future as a major variable in determining therapeutic aggressiveness.

Granulocyte-macrophage colony-stimulating factor associated with induction treatment of acute myelogenous leukemia: a randomized trial by the European Organization for Research and Treatment of Cancer Leukemia Cooperative Group.

PURPOSE: To assess the value of granulocyte-macrophage colony-stimulating factor (GM-CSF) for induction treatment of acute myeloid leukemia (AML), both for priming of leukemic cells and for acceleration of hematopoietic recovery. PATIENTS AND METHODS: GM-CSF was administered 5 micrograms/kg/d by continuous intravenous (i.v.) infusion during induction therapy with daunorubicin (DNR) (days 1 to 3) and cytarabine (ARA-C) (days 1 to 7). A total of 102 patients were randomized onto four arms, as follows: (1) GM-CSF 24 hours before and during chemotherapy (arm +/-); (2) GM-CSF after chemotherapy until day 28 or recovery of polymorphonuclear leukocytes (PMNs) (arm -/+);(3) GM-CSF before, during, and after chemotherapy (arm +/+); or (4) no GM-CSF (arm -/-). Stopping rules were applied in case of an initial WBC count greater than 30 x 10(9)/L or a secondary increase of circulating blast cells. Analyses were performed according to the intention-to-treat principle. RESULTS: The complete remission (CR) rates were 77% (arm -/-), 72% (arm +/-), 48% (arm -/+), and 46% (arm +/+). Patients randomized to receive GM-CSF after induction (arms -/+ and +/+) had a significantly lower CR rate (P = .008) and a trend toward accelerated recovery of neutrophils, but no fewer infections or induction deaths. The lower CR rate appeared to be related to an increased resistance rate, with persistent leukemia. The main side effects of GM-CSF were fluid retention and hypotension. CONCLUSION: GM-CSF administered during induction treatment of AML with a DNR/Ara-C combination did not provide any clinical benefit. Furthermore, there was a significant decrease in the CR rate with more persistent leukemia when GM-CSF was administered during the hypoplastic phase after the chemotherapy courses.

Fluorescence in situ hybridization analysis of minute marker chromosomes in leukemia with monosomy 7.

Monosomy 7 was detected in bone marrow cells from three patients, one with myeloid leukemia, and two others with myelodysplastic syndrome following previous chemotherapy. Fluorescence in situ hybridization (FISH), carried out with an alphoid DNA probe specific for chromosome 7 centromere, showed that a small marker chromosome present in the tumor cells' karyotype of the three patients, was derived from the missing chromosome 7. In two cases, the marker was a ring chromosome, whereas in the third case it was a tiny dot-like chromosome, unnoticed at first examination on R-banded metaphases. In the three cases, the marker was lost in a proportion of tumor cells. FISH experiments suggested that the marker centromere had undergone structural alterations, with a fluorescence pattern distinct from a normal one. On the whole, these data suggest that: firstly, leukemia-associated monosomy 7 results, in a proportion of cases, from a structural event rather than from simple loss of a whole chromosome 7; secondly, interpretation of interphase FISH must be cautious in monosomy 7 evaluation; and thirdly structural alteration of the chromosome 7 derivative alphoid DNA could explain its propensity to segregate unequally and to be lost at mitosis.

Treatment of patients with acute promyelocytic leukemia. The EORTC-LCG experience. EORTC Leukemia Cooperative Group.

Acute promyelocytic leukemia (M3) is, as one of the FAB subtypes of AML, included in the EORTC/GIMEMA AML-8A and 8B randomized trials. In these trials 1519 patients were included, 477 of them in non-Italian EORTC-LCG centers and 1042 in GIMEMA centers. A total of 80 patients were classified as M3 including 18 patients with M3-variant. Thirty-nine were male and 41 female. Ages ranged from 15 to 59 years; 25 (31.3%) of them were younger than 30, 34 (42.5%) between 30 and 45, and 21 (26.3%) older than 45 years of age. 56.3% of the patients had leukocytes less than 5 x 10(9)/l at the time of diagnosis vs. 24.9% of the patients belonging to the other FAB subtypes. Remission induction consisted of a standard protocol with 3 days daunorubicin and 7 days of cytosine arabinoside. Forty-three patients (53.8%) achieved a complete remission compared to 64.6% of the remaining AML patients. After salvage treatment this percentage increased to 70%, which is the same as for the other AML subtypes. Thirteen (16.3%) patients died during remission induction, mainly due to hemorrhagic complications. This percentage is significantly higher than the death rate (9.1%) in the other FAB subtypes of AML. All patients received one course of consolidation treatment. Post consolidation treatment could be either standard maintenance, intensive consolidation courses, autologous or allogeneic transplantation, according to the guidelines of the treatment protocols. At present, relapses almost all in the bone marrow, are seen in only 34.9% of the M3 patients, compared to 48.4% in the remaining AML patients. Disease-free survival for patients less than 45 years of age with the M2 and M3 subtypes was approximately 50% at 3 years compared to 30-40% for the other FAB subtypes. Despite the higher death rate during induction, the long-term survival results were better for M3 patients in comparison with the remaining AML patients. The projected survival at 3 years was 50% for M3 patients vs. 38% for remaining patients.

Occupational risk factors for acute leukaemia: a case-control study.

A case-control study has been performed for occupational risk factors of acute leukaemia, based on 185 cases more than 30 years old and 513 matched controls. There was a significant excess of polyvalent farming and electronic engineers among professions of cases, and, in addition of metal workers when considering the professions pursued for more than 5 years. The corresponding exposures were analysed through a detailed questionnaire, and assessed by an industrial hygienist after blinding the case-control status. The odds ratios (OR) were computed after adjustment on matching variables and prior chemo- or radiotherapy treatment, and after stratification for the level and total duration of exposure. There was no excess of professional exposure to ionizing radiation among cases. A significant relationship was observed between acute leukaemia and high or medium exposure to benzene, as well as over 10 years high or medium exposure to exhaust gas. In addition a significant relationship was observed with exposure to pesticides--insecticides and/or weed killers--and to electric and magnetic fields (EMF). The relationship with pesticides was significant when considering high or medium exposure to weed killers and more than 10 years exposure to both subtypes of pesticides. The relationship with pesticides and EMF remained significant when confounding factors were taken into consideration and after adjustment on co-exposure to benzene. The cytological studies showed that acute leukaemias following exposure to benzene (high or medium) and to EMF were only of myelogenous subtypes, whereas those following exposure to pesticides were divided between lymphoblastic and myeloblastic subtypes. Cytogenetic studies failed to show increased frequency of chromosomal abnormalities, as described in acute leukaemias secondary to anti-cancer treatments. Our study adds credence to the hypothesis that pesticides and EMF are leukaemogenic agents, together with benzene.

T-cell depletion does not affect long-term human bone marrow culture.

Bone marrow long-term cultures were studied for 8 weeks in four experiments before and after T-cell depletion with a mixture of monoclonal antibodies directed against CD2, CD5 and CD7. No differences in cellularity or granulomonocytic progenitor numbers appeared during the period of culture. We conclude that in vitro treatment in itself is not likely to explain non-take or graft rejection associated with T-depleted bone marrow transplantation.

Another case of trisomy 4 with double minute chromosomes in acute non-lymphocytic leukemia.

A further case of trisomy 4 with double minute chromosomes in acute non-lymphocytic leukemia is reported. The non-random association between these two cytogenetic abnormalities is reinforced. A possible relation with environmental exposure is discussed.

Pentasomy 13q in a case of acute myelogenous leukemia (Mo).

A case of acute myelogenous leukemia Mo FAB subtype with a pentasomy 13q (associated with a trisomy 19 in a subclone) in the initial bone marrow metaphase cells is reported. The pentasomy 13q is the result of the presence of double isochromosome 13q and one normal chromosome 13. In our case, this abnormality had a poor prognosis.

In vitro effect of rGM-CSF on proliferation and maturation of leukemic cells from patients with acute myelogenous leukemia.

We investigated the effect of rGM-CSF on the proliferation/differentiation balance of the leukemic cells maintained in liquid cultures during 7 days, from 16 patients with acute myelogenous leukemia (AML). Cell proliferation was measured by tritiated thymidine (3HT) incorporation, and by the plating efficiency (PE) observed after 7 days of culture. Differentiation was measured by the ability of cells to reduce nitroblue tetrazolium (NBT) and by the percentage of immature myeloid cells. After 7 days of culture without rGM-CSF, an increase of 3HT incorporation (p = 0.01 compared to control) was observed in 8 cases. In these patients, an absence of PE was noted in only one case. Among the 8 patients with decreased 3HT incorporation (p = 0.01 compared to control), 6 exhibited functional maturation (increase of % of NBT + cells, p = 0.01), and 4 showed no PE. Seven days exposure to 50 ng/ml rGM-CSF increased the leukemic cell proliferation in 9 cases, induced complete functional differentiation in 4, and enhanced the CFU-L recovery in 6 cases. These effects were mainly observed in the "proliferative" group, where 7 of the cases had an increase of 3H-T. However, two of the "non proliferative" cases were also stimulated by GM-CSF. An absence of proliferation was generally accompanied by functional maturation.

Early cytoreduction: a major prognostic factor in adult acute lymphoblastic leukemia.

Prognostic factors in acute lymphoblastic leukemia (ALL) are used for treatment stratification of ALL. Definition of simple parameters such as the presence or absence of peripheral leukemic cells after one week of treatment could help for stratification. A retrospective study was conducted on 79 previously untreated adult patients with ALL followed in the Hematology department of Hotel Dieu from 1981 to 1991. 84% of patients achieved complete remission (CR), 7% were refractory to induction treatment, and 7 patients (9%) died during the first month after diagnosis. After multivariate analysis the only independent statistically significant factors for achieving CR were the absence of peripheral blast cells at day 7 (PBC D7) (p = 0.009) and age (< 50 years) (p = 0.03). For CR duration the same independent statistically significant factors were found (PBC D7 = 0 versus > 0, p = 0.008; and age < or > or = 30 years, p = 0.045). The PBC D7 value was more significant when circulating blast cells were present at diagnosis. In patients with more than 50,000 PBC at diagnosis, the 10- years event free disease was 62% +/- 20% when PBC were absent at day 7 versus 0% when PBC were present (p < 0.002). All 20 patients with prolonged DFS had PBC D7 = 0 achieving CR by 28 days. The persistence of PBC at Day 7 could be used as a factor to identify a subgroup of poor prognosis adults with ALL.

Treatment of chronic myelogenous leukemia in blast crisis and in accelerated phase with high- or intermediate-dose cytosine arabinoside and amsacrine.

Twenty-two patients (mean age 41 years) in blast crisis or accelerated phase (AP) of chronic myelogenous leukemia (CML) were treated with cytosine arabinoside (Ara-C) 500 mg/m2 [intermediate dose] or 1000 mg/m2 [high dose] twice a day for 6 days and amsacrine (AMSA) 120 mg/m2 for 3 days. Twenty-one cases were of myeloid type and one was a lymphoid BC. The mean duration of aplasia (neutrophils < 0.5 x 10(9)/l) was 21.5 days. Four patients (18%) died of infection during aplasia and minor toxicities were noted for the remainders. Nine patients (41%) achieved a complete remission (CR) and 4 (18%) a partial response. Various additional therapies were proposed after induction treatment including allogeneic bone marrow transplantation (2 patients), Ara-C and AMSA maintenance or other regimens with or without alpha-interferon (9 patients). Median survival for the entire cohort was 20 weeks (wks), significantly superior for complete responders (37 wks) than for others (7 wks) (p = 0.008). In this study, age, sex, initial platelet or basophil counts, interval between diagnosis of CML and blast crisis were not predictive of response. Although inducing a high CR rate and associated with acceptable toxicity, this regimen did not improve the survival of patients with BC or CML, strengthening the need for alternate approaches to be defined.

Evaluation of carboplatin as a single agent in highly refractory acute myeloid leukemia.

Thirteen patients (pts) with highly refractory acute myeloid leukemia (AML) 10 pts with de novo AML and 3 with blast crisis of chronic myeloid leukemia were treated with carboplatin (CP) 150 mg/m2/day through continuous IV infusion for 7 consecutive days. Seven of them received CP at least as third or more line therapy after a median duration of the disease of 26 weeks. None achieved a complete remission but a good hematologic response, with disappearance of circulating blast cells along with correction of bone marrow failure, persisting for 3 months was obtained in one patient and correction of hyperbasophilemia was observed in another with blast crisis of chronic myelogenous. Myelosuppression was the most consistent toxic effect. Two deaths occurred, one from renal acute failure and the other from sepsis. Median survival after CP was 8 weeks (range 4 days-11 months) and the majority of patients were able to return home. When used as a single agent and with the dose-schedule used in this study, CP does not appear effective in refractory AML. Other studies are necessary to assess its role at an higher dose or in combination with other agents in earlier phases of the disease.

[Myelodysplasia and tuberculosis]. / Myélodysplasie et tuberculose.

We report here the case of a patient with myelodysplasia and concomitant tuberculosis. Refractory anemia with blasts excess diagnosis was based upon morphological and cytogenetical criterias (del 20q), and tuberculosis was diagnosed on a cervical lymph node biopsy. Hematological data remained stable without any specific treatment for several months, cell counts even normalized under antituberculosis tritherapy. Clinical and hematological worsening appeared 3 years later, 1 year after discontinuation of antituberculosis therapy. It was characterized by progressive bone marrow failure and transformation in acute myeloid leukaemia. Concomitantly tuberculosis relapsed. The association of antituberculosis therapy and polychemotherapy (daunorubicine and aracytine) did not allow to obtain a hematological remission. The relationship between tuberculosis and myelodysplasia is discussed.

[Acute nonmyeloid leukemia in adults. Study of membrane antigens and terminal transferase. Diagnostic and prognostic value]. / Leucémies aiguës non myéloïdes de l'adulte. Etudes des antigénes de membrane et de la terminal transférase. Valeur diagnostique et pronostique.

Between January, 1982 and April, 1983, 92 adult patients with acute leukaemia were investigated in our department. According to classical criteria (cytology and optical cytochemistry), 34 were classified as "non-myeloid". These were further tested with a panel of monoclonal antibodies against cell membrane, including ALB1-2 (anti-CALLA), ALB6 (anti-p24), ALB7-8-9 (BA1), OKT and HLA DR; they were also tested for E rosettes, Slg and terminal transferase (TDT). When all these markers but HLA DR were negative, patients were investigated for ultrastructural peroxidases which were found to be present in 2 cases. Among all non-myeloid leukaemias, 18 (56%) were CALLA-positive and TDT-positive acute lymphoblastic leukaemias (ALL), 2 (6%) were ALL T, 7 (22%) were CALLA-negative and TDT-positive ALL and 5 (16%) were acute leukaemias null for our markers, a phenomenon the significance of which is discussed. Patients with the CALLA-negative TDT-positive phenotype were peculiar with regard to age (mean: 50 years), female predominance, L2 cytological pattern according to the FAB classification, good prognosis (complete remission in 100% of the cases) and median survival (p less than 0,03).

[The prognosis factors of subacute and chronic myelo-monocytic leukemias (author's transl)]. / Les éléments du prognostic des leucémies myélo-monocytaires subaiguës et chroniques.

The myelo-monocytic leukemias (MML) can be of acute, subacute or chronic type, according to the level of bone-marrow blast cells and to the spontaneous course. We have compared the clinical, hematological and biological characters of 11 cases of subacute MML-defined by a survival of less than 12 months - to 20 cases of chronic MML. Anemia, hyperleucocytosis, monocytosis, number of circulating granulocytes and blast cells, level of bone marrow blasts, were superior in acute MML, with significant difference for hemoglobin, circulating and bone-marrow blasts. A high urine lysozyme output, a decrease of granulo-monocytic colonies after bone-marrow culture on semi-solid media were further arguments in favor of the subacute type. This variety, which evolves into acute MML in near 50 p. cent of cases appears consequently as the first step of an acute leukemia, or represents a very close aspect. Chronic MML on the contrary, although with still 30 p. cent blastic transformation, is characterized by a steady course, with a median survival of 3.3 years, and number of untreated patients surviving many years beyond this median time.

[Contribution of cytochemistry and delayed hypersensitivity skin tests in the diagnosis and prognosis of chronic lymphoproliferative syndromes]. / L'apport de la cytochimie et des tests cutanés d'hypersensibilité retardée dans le diagnostic et le pronostic des syndromes lymphoprolifératifs chroniques

Cytochemistry is disappointing in lymphoproliferative syndromes for it does not permit one to classify the various diseases with certainty. In the early stages, if the three indices are lowered, the prognosis seems poorer. A study of glucuronidase permits, in hyperlymphocytosis, one to differentiate benign from malignant lymphocytes, but does not permit one to differentiate from one another, the other chronic lymphoproliferative syndromes. The acid phosphatase is interesting in the study of hairy cell leukemia. Finally, it was not possible to distinguish chronic lymphoid leukemia from leukemia with lymphosarcomatous cells, nor from the cytochemical point of view nor using tests for delayed hypersensitivity.

Acute myelogenous leukaemia in the elderly: retrospective study of 235 consecutive patients.

A retrospective analysis was performed on 235 elderly acute myelogenous leukaemia (AML) patients aged 60 years or more, consecutively admitted to a single haematological department during a 10-year period from 1980 to 1989. 46% of patients received only conventional induction chemotherapy. The rate of inclusion in EORTC cooperative clinical trials was significantly lower than for younger patients despite specific protocols proposed for the elderly since 1983, thus confirming the important selection bias of most published series on elderly AML patients. Compared with treatment results in patients < 60 years, complete remission (CR) rate was lower (33.3% v 65.4%, P < 0.0001), with a marked drop in patients older than 70, and induction death rate was higher (21.3% v 12.5%, P = 0.04). Intrinsic characteristics of leukaemic cells, especially expression of the MDR1 gene, in vitro growth of the leukaemic clonogenic cells and sensitivity to daunorubicin-(+)cytosine arabinoside, did not differ according to age, except that there was a higher incidence of previous myelodysplastic syndromes and a lower incidence of good prognostic cytogenetics in the elderly patients. Thus, treatment failure in elderly AML patients appears to be mainly due to host-related factors (especially performance status and age < or > or = 70 years), and to inadequate treatments. Some elderly patients may have been undertreated because of the planned anthracycline dose reduction, resulting in a higher rate of 'resistant' AML, i.e. patients surviving the induction period without entering into CR, than in younger patients (45.4% v 22.1%, P < 0.0001). 11 patients (4.7%) with untreated or 'resistant' AML survived more than 1 year, while receiving only supportive care. These slowly progressive AML patients were characterized by a good performance status, and lower circulating blast cells and bone marrow blast counts.

[Biermer's disease without anemia, nor megaloblastosis. Value of the study of antibodies, vitamin assays, and the dU suppression test for the early diagnosis]. / Maladie de Biermer sans anémie, ni mégaloblastose. Intérêt de la recherche des anticorps, des dosages vitaminiques, et du test de dU suppression pour le diagnostic précoce.

In two patients, a diagnosis of pernicious anemia was made without anemia, megaloblastosis and even macrocytosis. The diagnosis of pernicious anemia was suggested by the autoimmune abnormalities which are frequently associated with this disease: goitre with anti-thyroid and anti parietal cell antibodies, and besides this, idiopathic thrombocytopenic purpura in the first patient, vitiligo in the second. The morphological abnormalities were limited to slight macrocytosis in the first patient and to hypersegmentation of polymorphonuclear leucocyte in both. Vitamin B12 deficiency was demonstrated by serum assay as well as deoxyuridine suppression test ("dU suppression"). The diagnosis was confirmed by demonstration of atrophic gastritis, failure of secretion of Intrinsic Factor and Schilling test. These observations show that the Addison-Biermer disease can be detected early in persons at high risk by looking for vitamin B12 deficiency and specific antibodies.

[Cytological classification of adult acute leukemia (author's transl)]. / Classification cytologique des leucémies aiguës de l'adulte

134 cases of acute leukemia in adults were classified according a "double blind" cytological diagnosis, a cytochemical study, and lysozyme assay. Each type has distinct characters, allowing a good nosological definition. The histochemical methods, as well as lysozyme assay are usually unnecessary, particularly in well differenciated myeloblastic, promyelocytic or myelo-monocytic acute leukemia. They are, on the contrary, frequently useful in poorly differenciated myeloblastic or monoblastic leukemia. But, they cannot help to solve every problem in cytological diagnosis: there are still 10% of undifferenciated acute leukemia, the lymphoblastic acute leukemia are not clearly defined and 4% of cases of acute leukemia have atypical characters leading to difficulties in cytological classification. The need for new methods and markers is emphasized.