Correlation of MDR1/P-170 expression with daunorubicin uptake and sensitivity of leukemic progenitors in acute myeloid leukemia.

Prior studies have shown that multidrug resistance gene products may be detected in acute myeloid leukemia (AML) cells, and are associated with poor response to therapy. We studied whether P-170 expression was associated with in vitro daunorubicin (DNR) accumulation and sensitivity of leukemic clonogenic cells (CFU-L) to DNR in 16 newly diagnosed AML samples. P-170 expression was assessed by indirect immunofluorescence using the monoclonal antibody MRK16. DNR cellular content was measured by flow cytometry after short incubation with increasing concentrations of DNR, and was not correlated with P-170 expression, although there was a trend for higher values in P-170-negative samples. The sensitivity of CFU-L was studied in a semisolid culture assay by calculating the dose of DNR inhibiting the growth of 90% of CFU-L (D90). The D90 was significantly higher in P-170-positive than in P-170-negative samples (mean = 1.68 +/- 0.42 microgram/ml versus 0.97 +/- 0.35 micrograms/ml respectively, p less than 0.005). Eight of 9 cases achieving complete remission (CR) after intensive chemotherapy were P-170-negative, whereas 7 of 7 nonresponders were P-170-positive (p less than 10(-5)). D90 was significantly lower for patients achieving CR than for those who did not achieve CR (1.12 +/- 0.55 micrograms/ml versus 1.59 +/- 0.37 micrograms/ml, p = 0.04). It is concluded that P-170 expression is correlated with in vitro resistance of clonogenic cells to DNR and may be one mechanism of resistance to chemotherapy.

Specific detection of monocytic lysozyme within normal and leukemic cells.

A murine monoclonal antibody against human lysozyme (AHL MoAb) was produced and tested on normal and leukemic monocytes using flow cytometry. The antibody gave a positive reactivity on normal monocytes permeabilized by saponin (82% to 98% of positive cells) and a negative reactivity on normal permeabilized neutrophils. This monocyte-specific reactivity had not been observed using a polyclonal antibody. Nevertheless, immunoblotting detected lysozyme in both monocyte and polymorphonuclear leukocyte (PMNL) lysates. The AHL MoAb, in the presence of lysozyme substrate (Micrococcus lysodeikticus cell walls), strongly inhibited the enzymatic activity. Flow cytometric analysis of leukemic cells isolated from patients suffering from different subtypes of acute myeloid leukemia (French-American-British [FAB] classification FAB M1-5) showed a highly significant positivity of FAB M5 for lysozyme compared with the other subtypes. The present results were consistent with the detection of a lysozyme epitope by AHL MoAb located near the catalytic site in monocytes. The same epitope was probably masked in PMNL granules.

Specific binding of anti-N-acetyllactosamine monoclonal antibody 1B2 to acute myeloid leukaemia cells.

1B2 is an IgM monoclonal antibody binding to glycoconjugates bearing the terminal N-acetyllactosamine structure. It agglutinates human erythrocytes. Various cell lines, peripheral blood leucocytes, normal marrow and blast cells from 179 acute myeloid leukaemia (AML) and 11 acute lymphoblastic leukaemia (ALL) patients were tested for reactivity with 1B2. Myelomonocytic (CFU-GM), erythroid (BFU-E), mixed (CFU-GEMM) and leukaemic (CFU-L) progenitor cells were tested in clonogenic assays. Granulocytes, monocytes, myeloid cell lines and 152 out of 179 AML were positive. All FAB subtypes were equally recognised. Lymphocytes, T-cell and Burkitt's cell lines, and 10 of 11 ALL samples were negative. 1B2 inhibited partially day 7 CFU-GM, whereas it was not toxic for BFU-E, CFU-GEMM and day 14 CFU-GM. Leukaemic clonogenic cells were killed in 33 out of 36 AML (more than 40% growth inhibition). 1B2 identifies the more mature steps of myeloid differentiation. It may be useful in the diagnosis of AML, and is a candidate for remission marrow purging before autologous transplantation.

Clinical significance of multidrug resistance P-glycoprotein expression on acute nonlymphoblastic leukemia cells at diagnosis.

To evaluate the clinical value of the expression of multidrug resistance P-glycoprotein (P-170) on the surface of acute nonlymphoblastic leukemia (ANLL) cells, we analyzed specimens from 150 newly diagnosed patients for staining with MRK16, a monoclonal antibody (MoAb) that binds to an external epitope of P-170. Other surface markers (CD13, CD14, CD15, and CD34) were studied by the same technique. A marker was considered positive when 20% or more cells were stained. Of 150 samples, 71 were P-170-positive. These cases did not differ from P-170-negative cases with regard to age, sex, initial white blood cell (WBC) counts, or French-American-British (FAB) type (except for M3 ANLL, which were more frequently negative). However, leukemias arising from previous myelodysplastic syndrome (MDS) and therapy-induced leukemias were more frequently P-170-positive. CD34 and P-170 expression were significantly associated. All patients were treated by intensive chemotherapy. Complete remission (CR) rates were significantly lower in P-170-positive (23/71, 32%) than in P-170-negative cases (64/79, 81%) (P less than 10(-5)). CD34 positivity was also associated with a low remission rate (P less than 10(-5)). Survival was shorter for P-170- and CD34-positive patients (P less than 10(-5)). The prognostic value of both markers was confirmed in multivariate analysis. CR duration was also shorter for P-170-positive cases, but the difference is less significant (P = .05). It is concluded that P-170 analysis may be an important tool for predicting the outcome of intensive chemotherapy in ANLL patients.

CD1-reactive leukemic cells in bone marrow: presence of Langerhans cell marker on leukemic monocytic cells.

Langerhans cells originate in bone marrow and probably belong to the monocyte-macrophage lineage. CD1 is a specific marker of Langerhans cells. By immunofluorescence and immunoelectron microscopy, CD1a antigen and myeloid markers (CD11, CD13, CD14, CD15, CD33, HLA-DR) were studied in 53 cases of acute myeloid leukemias (AML) and 3 acute lymphoblastic leukemias (ALL). The 11 ANLL without monocytic component were CD1a negative. 2/5 of acute myelomonocytic leukemias (AML4) and 9/37 of acute monocytic leukemias (AML5) were positive. All 3 ALL were negative. No correlation was found between CD1a and myeloid markers. CD1a+ AML did not differ from CD1a- AML with regard to cytogenetics or response to therapy. The CD1a positive cells may originate from an abnormal proliferation of CD1a positive cells which are present in bone marrow and which may differentiate into Langerhans cell precursors.

Myeloid surface antigen expression in adult acute lymphoblastic leukemia.

The expression of myeloid surface markers was investigated in 41 cases of untreated adult acute lymphoblastic leukemia (ALL). Nineteen cases (46%) reacted with at least one myeloid monoclonal antibody (CD15 in 16 cases, CD13 in 10 cases, CD14 in five cases, and CD33 in four cases). Double-staining confirmed the coexpression of myeloid and lymphoid markers. In addition, 35 samples were tested for CD34 expression. Fourteen of the 17 myeloid-positive cases tested were positive for CD34 vs. eight of 18 negative cases (p less than 0.05). A t(9;22) translocation was found in eight cases, and a t(4;11) translocation in two cases, all expressing CD34 and myeloid antigens. These findings confirm the high frequency of myeloid markers on the surface of adult ALL blasts, and suggest that these leukemias may originate in a poorly differentiated precursor cell with mixed differentiation capacities.

Surface marker expression in adult acute myeloid leukaemia: correlations with initial characteristics, morphology and response to therapy.

The clinical significance of surface markers was investigated in 145 cases of acute myeloid (AML) or undifferentiated leukaemia (AUL), using a panel of six monoclonal antibodies directed to NHL-30.5 antigen (expressed on poorly differentiated myeloid cells), CD13, CD14, CD15, CD33 and CD34 antigens. Expression of CD14 was correlated with higher leucocyte count, higher serum lactate dehydrogenase level and presentation with extramedullary disease. There was no strict correlation with the French-American-British classification. However, the expression of CD14 was associated with monocytic subtypes. CD15 was mainly expressed in M2 and M3 subtypes, and NHL-30.5 and CD34 antigens in AUL and M1 leukaemias. All patients were treated with the same intensive induction treatment. Staining by three antibodies had a prognostic value. The complete remission (CR) rates were 38% (26/68) in NHL-30.5-positive versus 75% (62/77) in NHL-30.5-negative cases (P less than 10(-5), 50% (37/74) in CD34-positive versus 72% (51/71) in CD34-negative cases (P = 0.007) and 70% (77/110) in CD15-positive versus 31% (11/35) in CD15-negative cases (P less than 10(-4). Expression of NHL-30.5 and CD34 antigen was associated with shorter survival (P less than 10(-3) and P less than 10(-2) respectively), whereas survival was longer in CD15-positive cases (P less than 10(-3). In multivariate analysis, expression of NHL-30.5 antigen, absence of CD15, and high LDH level were associated with poor survival. CR duration was not influenced by any of the factors studied, including antigen expression. These results suggest that leukaemias with less differentiated phenotype have a lower response rate to induction treatment.

Acute leukaemia following high-dose chemoradiotherapy with bone marrow rescue for ovarian teratoma.

A case of acute leukaemia following intensive chemo- and radiotherapy for solid tumour is reported. A 15-year-old girl received four courses of chemotherapy with adriamycin, cyclophosphamide and cisplatinum after the surgical diagnosis of ovarian immature teratoma. An intensification treatment was performed with cyclophosphamide and total body irradiation, followed by marrow rescue. The diagnosis of myelomonocytic acute leukaemia was performed 8 months later. The patient died after a transient improvement following a treatment by hydroxyurea. The roles of the primary tumour, of the chemotherapy and of the marrow transplantation in the occurrence of leukaemia are discussed.

Expression of a B-lymphoid differentiation antigen (CD 19) on acute non-lymphoblastic leukaemia cells.

The expression of CD 19 (B4) surface antigen was investigated in 41 cases of acute myeloid leukaemia (AML), and 8 cases of acute undifferentiated leukaemia (AUL). Diagnosis was carried out by usual cytological and cytochemical means, and leukaemias were classified according to the FAB classification. CD 19 antigen is thought to be expressed on B lymphoid cells only. However, 2 cases out of 9 FAB M1, M2 and M3 subtypes were weakly stained (20 to 25%) by B4 antibody; 7 out of 11, and 17 out of 18 cases of M4 and M5 subtypes were markedly stained (25 to 70% of cells). 3 cases of AUL were also stained. Moreover, B4 expression was often correlated with expression of monocyte-associated antigens, as recognized by CRIS6, My9, KiM1 and UCHM1 antibodies. It is concluded that B4 antigen is probably expressed before B differentiation and is retained in early stages of monocytic differentiation.

Surface markers in acute non-lymphoid leukemia: analysis with a panel of 36 monoclonal antibodies.

The reactivity of a panel of monoclonal antibodies was studied in fifty-four cases of acute myeloid (AML) or undifferentiated (AUL) leukemias. Thirty-six antibodies from the Myeloid section of the Second Workshop on Human Leukocyte Differentiation Antigens were used in an indirect immunofluorescence assay. The antibodies could be classified into three groups recognizing respectively granulocytic, monocytic or granulomonocytic leukemias. Most antibodies stained erythroblastic and megakaryoblastic leukemias. In each group, it was possible to define antibodies staining either the less differentiated forms (FAB M 1 and M 5 a) or the more differentiated forms (M 2, M 3, M 4 and M 5 b). Six out of eight AUL were stained by some of the antibodies (mainly from the monocytic group). However, a heterogeneity of stainings in a same blast population was observed.

[Prolonged survival in adults with acute myeloid leukemia treated with BCG immunotherapy]. / Longue survie dans les leucémies aiguës myéloïdes de l'adulte sous immunothérapie par le B.C.G.

Between november, 1974 and june, 1977, 64 adult patients with acute myeloid leukaemia in remission were divided at random into two treatment groups: chemotherapy alone (n = 32) or chemotherapy plus B.C.G. (n = 32). Fresh B.C.G. from the Pasteur Institute, Paris, was administered once a week, two or three times per month, in the intervals between chemotherapy cycles, using the Heaf-gun method. The duration of the first remission was significantly increased in the group receiving B.C.G. (median duration 685 days versus 390 days: p = 0.03 on log rank test) as was the overall survival (median duration of survival 1.471 days versus 554 days: p = 0.01). Second remissions were more frequent in the B.C.G. group (79% versus 42%; p = 0.014). In addition, prolonged survival was more frequent in the B.C.G. group, where 12/32 patients remained in first remission between 3 and 6 years, as against 5/32 in the chemotherapy group.

Prognostic value of mouse red cell rosette formation in chronic lymphocytic leukaemia.

Mouse red cell (MRC) rosette formation was studied in 20 malignant haemopathies. Only cells from chronic lymphocytic leukaemia (CLL) and hairy cell leukaemia (HCL) formed MRC rosettes with a high frequency. However, cells from 2 cases of CLL could not be demonstrated to form MRC rosettes, although they were undoubtedly found to be of B origin. These 2 cases presented as typical CLL, but they were observed to transform into a sarcomatous type after 5-6 months of evolution, suggesting that the study of MRC rosette formation might be of prognostic value in CLL.