Simultaneous analysis of DNA and cellular proteins provides information on cell proliferation and metabolism. Cellular protein content coupled with nuclear geometric parameters can be used to evaluate cellular maturation and differentiation. In this study, leucoblasts from 50 cases of adult acute myeloid leukemia were analyzed by flow cytometry, and semiautomatic morphometry was performed on bone marrow smears. Ethanolfixed bone marrow blast cells were stained for DNA with propidium iodide (PI) and for proteins with fluorescein isothiocyanate (FITC). On the resulting FITC versus PI histograms we defined the cells with low protein content which are associated with a nonproliferating subpopulation (LPC fraction). Low protein content fraction and S-phase are correlated (p < 0.01). The LPC fraction values are more dispersed than S-phase values and thus should indicate more clearly eventual differences between cellular populations. This hypothesis has been tested with the prognostic significance of cell-cycle variables: The LPC fraction was significantly higher in the complete remission group than in the other (p < 0.01), while S-phase did not show any difference.The peak value of the protein content histograms is significantly lower in the granulocytic leukemias (Ml, M2, M3) than in the leukemias with a monoblastic component (M4, M5). Furthermore, we showed that the differentiation and the maturation of the myeloid blast cells modify the nuclear size. The combination of these two parameters provides useful information for cytological classification.Key terms: Acute leukemia, cell cycle, analytical cytometry, PI + FITC staining Acute myeloid leukemia (AML) displays a broad spectrum of morphological and functional cellular abnormalities as well as a large diversity in clinical evolution and in therapeutic response. In contrast to acute lymphoid leukemia (ALL), for which immunological markers allow a precise evaluation of the functional differentiation, AML is usually dissected and analysed by means of morphological criteria (7,221, cytokinetic characterization (1,3), or cytogenetics (6).Cytology usually indicates that the differentiation and the degree of maturation of blastic cells can be assessed by cell size and protein content. MATERIALS AND METHODSPatients Fifty patients aged 18 to 75 years with AML were studied before chemotherapy. None of these patients had any record of dysmyelopoiesis and none had received any chemotherapy prior to this study.Acute myeloid leukemias were classified according to differentiation and maturation criteria using the FrenchAmerican-British (FAB) classification (7): M1: 11 cases, M1-M2: 2 cases, M2: 10 cases, M3: 7 cases, M4: 13 cases, Address reprint requests to Dr.

Serum beta 2 microglobulin levels, measured by radioimmunoassay (Phadebas test), were found increased in acute myeloid leukemias at diagnosis. Serum beta 2 microglobulin levels were significantly higher in patients with monocytic leukemias (13 patients, M4-M5 FAB classification) than in those with other cytological types (18 patients). Beta 2 microglobulin levels at diagnosis were correlated with serum lysozyme levels, but they were not correlated with blood blast counts, serum LDH and ferritin levels. 195 serum beta 2 microglobulin measurements were made serially in 30 patients with acute myeloid leukemias in first remission. Compared to values at diagnosis, beta 2 microglobulin levels in remission were significantly decreased. Out of 30 patients in remission 12 had increased serum beta 2 microglobulin levels (greater than 3 mg/l). Serial measurements were not predictive for relapses.

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A woman affected by acute myeloblastic leukemia was grafted with HLA A, B and D compatible rhesus-positive bone marrow from her brother. Before grafting, she had anti-D alloantibodies (1/512 IAT, 2.9 μg/ml). To prevent the destruction of donor red blood cells, four plasma exchanges and a conditioning regimen (total-body irradiation 800 rad, cyclophosphamide, methotrexate) were carried out to decrease anti-D from 2.9 to < 0.02 μg/ml on day 0. The anti-D level was 0.8 μg/ml on day 12 and was decreased to 0.2 μg/ml by eight plasma exchanges until day 35. Anti-D antibodies were undetectable with Lalezari’s technique on day 45. Engraftment was obtained on day 25 (3,000 leukocytes/mm³ and 50% erythroblasts in bone marrow). The patient died from aspergillosis and graft-versus-host disease on day 54. This observation shows that an engraftment of rhesus-positive bone marrow in a recipient with anti-D antibody is possible.