Prognostic importance of Sudan Black positivity: a study of bone marrow slides from 1,386 patients with de novo acute myeloid leukaemia.

Analysis of bone marrow slides from 1,386 patients entered into the Medical Research Council's 8th and 9th trials in Acute Myeloid Leukaemia confirmed that features associated with differentiation in blast cells, in particular increasing Sudan Black (SB) positivity, were the most important morphological features for predicting remission achievement (P = 0.002) and hence survival (P less than 0.0001). SB positivity was also weakly predictive of remission duration (P = 0.05). A low complement of maturing granulocytes was associated with early induction death and a high percentage of blasts with shorter remissions. The few patients with acute promyelocytic leukaemia (FAB M3) had a high haemorrhagic death rate during induction and a low relapse rate. Apart from this, lineage involvement was not predictive of outcome. Multiple lineage leukaemias, in particular those with megakaryocytic and/or erythroid involvement, which had been reported previously to have a poor prognosis, did not have any worse remission rates in this series. When more than one cell line was involved, no combination with particularly good or poor prognosis could be identified. Multivariate analysis suggested that percentage SB positivity was adequate on its own to divide granulocytic leukaemias into poorly differentiated (less than 50% SB +ve) and well-differentiated groups (50% or more SB +ve) without the need for further measurements. This simple and reproducible test was strongly predictive of resistant disease but not of induction deaths. It was of considerably greater prognostic value--and was less open to inter-observer disagreement--than the FAB criteria which are usually used to classify granulocytic lineage leukaemias into the M1 and M2 subgroups. It is proposed that greater than or equal to 50% of blasts with SB positivity should replace blasts greater than 10% of maturing myeloid cells for this sub-categorization between M1 and M2.

A new human plasma cell line, Karpas 620, with translocations involving chromosomes 1, 11 and 14.

We report here the establishment of a new cell line, Karpas 620 (K620), from the peripheral blood of an elderly woman with an IgG-kappa plasma cell leukaemia (PCL). The line has the same hypotetraploid karyotype as the fresh cells from the patient. The cultured cells have the ultrastructural appearance of plasma cells with abundant rough endoplasmic reticulum (RER) and secrete kappa light chain. They are positive for surface antigens HLA DR, and WR17 (CD 37) and negative for CD1, CD3, CD4 and CD8. Using high resolution (HR) cytogenetic analysis it has been possible to identify all the marker chromosomes including several rearrangements commonly seen in malignancies of B cell lineage. These are a 14q+ marker with a typical 'Burkitt' morphology der(14)(pter----q32.3::8q24.1----qter) but with no reciprocal 8q-, and three translocations involving chromosome 11 at q13 with partners other than chromosome 14, namely 1q32.1, 8q24.22 and 13q14.3. An earlier report of molecular studies on the DNA of K620 has shown a rearrangement near the region on 11q13 designated BCL-1 (Rabbitts et al. 1988). This is the first report of a rearrangement in the region of 11q13 in a cell line originating from a case of plasma cell leukaemia.

The significance of ultrastructural features in acute myeloid leukemia: a study of 220 patients entered into the Medical Research Council's ninth acute myeloid leukemia trial.

Samples of peripheral blood or bone marrow obtained before treatment from 220 patients entered into the Medical Research Council's ninth acute myeloid leukemia (AML) trial were examined by electron microscopy. Several ultrastructural features showed strong correlations with each other, with variants of AML defined by the Hayhoe classification scheme, and with FAB type. However, no ultrastructural feature was found either uniquely in association with any other or specifically in any variant of AML. Each ultrastructural feature was tested for association with achievement of remission, remission duration, and survival time. Only Auer rods were associated with a high remission rate (p = 0.01), but even this association was not quite conventionally significant when the analysis was not quite conventionally significant when the analysis was stratified for age (p = 0.055). Shorter duration of remission was associated with cytoplasmic projections (p = 0.04 stratified for age). Overall survival was worse when convoluted or lobed nuclei were present but only when stratified for age and Hayhoe type (p = 0.04). The possibility of testing combinations of ultrastructural features for correlation with diagnosis or prognosis is discussed but would require data from more than the present 220 patients to be meaningful.

Basophils in acute myeloid leukaemia.

Thirty four out of 750 patients entered into the Medical Research Council's (MRC) 9th Acute Myeloid Leukaemia Trial had more than 1% basophils (range 1-27%) often with bizarre granulation and primitive forms, a rare finding in this disease. Both normal and abnormal karyotypes were present including abnormalities of 6p, 12p, and the Philadelphia chromosome. Basophilia was found in both "monolineage" and "multilineage" leukaemias and the commonest French-American-British (FAB) classification group was M2, followed by M4. Basophilia did not seem to be associated with a worse prognosis, although cases with abnormalities of 6p died of disease that was resistant to first line conventional chemotherapy.

Effects of CAMPATH-1 antibodies in vivo in patients with lymphoid malignancies: influence of antibody isotype.

The CAMPATH-1 family of antibodies recognize an abundant glycoprotein expressed on virtually all human lymphocytes. All rat IgM and IgG antibodies of this specificity are lytic with human complement, but only IgG2b is active in antibody-dependent cell-mediated cytotoxicity (ADCC). We compared the ability of IgM, IgG2a, and IgG2b to deplete lymphocytes in vivo in two patients with prolymphocytic transformation of B-cell chronic lymphocytic leukemia (CLL). The IgM (CAMPATH-1M) produced transient depletion of blood lymphocytes with consumption of complement but had no effect on solid masses or bone marrow. Similar transient depletion of blood lymphocytes was noted with the IgG2a (YTH34.5). In contrast, the IgG2b (CAMPATH-1G) produced long-lasting depletion of lymphocytes from blood and marrow and improvement in splenomegaly but no detectable changes in complement levels. These differences probably reflect the importance of Fc receptor binding for effective clearance of target cells in vivo. We treated 16 more patients with a variety of lymphoid malignancies and noted consistent effects on blood lymphocytes, marrow infiltration, and splenomegaly. At this dose level, there was comparatively little improvement in affected lymph nodes or extranodal masses. Nevertheless, the in vivo lympholytic ability of CAMPATH-1G is very potent as compared with other monoclonal antibodies (MoAbs) and may have applications in therapy of lymphoid malignancies and as an immunosuppressive agent.

AML associated with previous cytotoxic therapy, MDS or myeloproliferative disorders: results from the MRC's 9th AML trial.

The outcome of treatment with standard first line therapy of 66 patients with acute myeloid leukaemia (AML) secondary to preceding chemotherapy (Group 1), a myelodysplastic state (Group 2) or a myeloproliferative disorder (Group 3) was analysed in relation to the preceding disorder, the cytogenetic pattern where available, and the cytology and cytochemistry of blood and bone marrow. The complete remission (CR) rate for the secondary AMLs was 36% (24/66), with 24% (16/66) dying in the induction period and 39% (26/66) having resistant disease. The CR rate was 25% (5/20) for Group 1, 42% (15/36) for Group 2, and 40% (4/10) for Group 3. Even after allowance for the generally older age of the secondary AML patients, they still had a significantly poorer CR rate than the de novo AMLs (P = 0.0004). The lower CR rate was chiefly due to resistant disease. Despite this, overall survival was not significantly worse for the secondary AML patients (P = 0.15). For the 36% that achieved remission, remission duration appeared similar to that of de novo cases. Of 62 cases with adequate cytology, 38 (61%) had evidence of erythroid and/or megakaryocytic dysplasia with a CR rate of 32% (12/38). The CR rate of these multineage leukaemias was not significantly different from that of the 24 (39%) who showed granulocyte/monocyte precursor involvement only, 42% (10) of whom achieved CR. The presence of features of differentiation within blast cells such as Auer rods or sudanophilia (greater than 50% positive blasts) was associated with a higher remission rate 47% (18/38) than that of poorly differentiated cases 17% (3/18) (P = 0.04) and thus appeared to be a more important determinant of CR achievement than was lineage involvement. Cases with a normal karyotype had a 33% (7/21) CR rate, while those with chromosomal abnormalities had a 37% (9/24) CR rate. Only 12 of the 45 cases with adequate cytogenetic analysis showed deletions or monosomies involving chromosomes 5 or 7, and seven of these were in Group 1.

A modern classification of the acute non-lymphoblastic leukaemias.

Acute leukaemias have traditionally been classified according to the nature of the predominating cells as judged by cytomorphology and cytochemistry. A codification of this classification into M1-M7 for myeloid cases (AML), proposed by the FAB group of haematologists, has been used extensively in the past decade. Some criticisms of this codification are presented and other approaches to classification are discussed. Among these are included the potential importance of multiple lineage expression, measurements of cell differentiation using cytochemical criteria, surface antigenic differences as demonstrated by the use of monoclonal antibodies and the growing contribution of cytogenetics. A simplified classification into broad groups is proposed. These groups may be further elaborated as appropriate.

Classification of acute leukaemias.

Acute leukaemias have traditionally been classified according to the nature of the predominating cells as judged by cytomorphology and cytochemistry. A codification of this classification into L1-L3 subdivisions for lymphoblastic cases (ALL) and M1-M7 for myeloid cases (AML), proposed by the FAB group of haematologists, has been used extensively in the past decade. Some criticisms of this codification are presented and other approaches to classification are discussed. Among these are included the potential importance of multiple lineage expression, measurements of cell differentiation using cytochemical criteria, the relevance of isoenzyme biochemistry of leukaemic cell populations, their surface antigenic differences as demonstrated by the use of monoclonal antibodies and the growing contribution of cytogenetics. The development and complexity of combined classifications which attempt to incorporate most of these features is illustrated. A simplified classification into broad groups is proposed, ALL being divided by surface antigenic and cytogenetic criteria and AML by morphology and cytochemistry. These groups may be further elaborated as appropriate.

Two further cases of acute myeloid leukemia with trisomy 4.

Two cases of acute myeloid leukemia (FAB M1 and M2) with trisomy 4 are described. Morphologic abnormalities were confined to the granulocytic monocytic lineage with no evidence of erythroid or megakaryocytic involvement. One of the cases, who also had trisomy 13, presented initially with skin infiltration without bone marrow involvement.

A Ki-1 (CD30)-positive human cell line (Karpas 299) established from a high-grade non-Hodgkin's lymphoma, showing a 2;5 translocation and rearrangement of the T-cell receptor beta-chain gene.

We describe the characterization of a new human cell line, Karpas 299 (K299), established from blast cells in the peripheral blood of a 25-year-old white man. His illness, which began with enlarged occipital and axillary nodes and weight loss, ended after 7 months with generalized lymphadenopathy, pleural effusion, and bone marrow involvement. A lymph node biopsy showed a large cell lymphoma mainly sinusoidal in distribution. The blast cells with pleomorphic nuclei resembled primitive histiocytes. The cells, which expressed the T-cell-associated markers CD4 and CD5, were positive for HLA-DR, epithelial membrane antigen, and CD30 (Ki-1 antigen). The karyotype was aneuploid and included a translocation 2;5. The site of translocation on chromosome 5 (at 5q35.1) is in the region of the locus of the c-fms oncogene (receptor of the monocyte-macrophage colony-stimulating factor MCSF or CSF-1). The cell line Karpas 299 has the same karyotype and pattern of antigen expression as the patient's cells. Northern blot analysis of RNA showed an active rearrangement of the T-cell receptor beta-chain gene. This is to our knowledge the first Ki-1 antigen-positive line to be established from a case of non-Hodgkin's lymphoma.

Translocation 5;21 and interstitial deletion of chromosome 7 in a case of chronic myelomonocytic leukemia.

We report a case of chronic myelomonocytic leukemia with a translocation involving chromosome 5 and 21, namely, t(5;21)(q35.3;q22.1), and an interstitial deletion of the long arm of chromosome 7. High-resolution analysis at the 900-band stage has shown that the lesion in chromosome 7 is an interstitial deletion involving loss of the segments q22-q35 of the long arm of chromosome 7 and retaining the telomere.

Beneficial effect of heparin in the management of patients with APL.

115 patients with acute promyelocytic leukaemia (APL) were studied retrospectively to evaluate prognostic factors and assess therapeutic approaches, particularly the use of heparin in the management of disseminated intravascular coagulation (DIC). The remission rate was 86% (30/35 patients) in those who received heparin and 49% (39/80 patients) in those who received no heparin (P = 0.0002). This difference in remission rates was accounted for by a marked decrease in the number of haemorrhagic deaths, especially those due to intracranial haemorrhage (ICH), in the heparin treated group. Other factors associated with a poor remission rate were prothrombin ratio (PTR) greater than 1.3 (P = 0.008), fibrinogen less than 1.5 g/l (P = 0.02) and WCC greater than 2.0 x 10(9)/l (P = 0.03).

Translocation (3;6)(q21;p21) in acute myeloid leukemia with abnormal thrombopoiesis and basophilia.

A case of acute myeloid leukemia (AML) with increased numbers of basophils and abnormal megakaryocytes was shown to have a t(3;6)(q21;p21) in the bone marrow cells. The morphology is described in detail and the case is discussed with reference to t(6;9)(p23;q34) and inv/ins(3)(q21q26) in AML. It is possible that increased numbers of basophils in AML may be associated with a translocation involving 6p21-6p23.

Complex translocation t(8;12;14) in a cell line derived from a child with nonendemic Burkitt-type acute lymphoblastic leukemia.

A cell line is described with a typical Burkitt lymphoma (BL) marker 14q+ due to the classical reciprocal translocation between chromosome #8 and #14 with breakpoints at 8q24.1 and 14q32.3. In addition, an interstitial piece from the long arm of 12(q24.1-q24.3) is inserted at the site of the exchange on chromosome #8, proximal to 14q32.3.

Cytochemical demonstration of 'platelet' peroxidase at the light microscope level.

A reliable and reproducible modified method was developed for the cytochemical demonstration of the so-called 'platelet' peroxidase at the light microscopy (LM) level. Interestingly, not only normal platelets and megakaryocytes showed the peroxidase activity, but also normal lymphocytes. As with the ultrastructural platelet peroxidase (PPO) method, myeloperoxidase (MPO) was also demonstrated in granulocytes. Peroxidase activity was exhibited by blast cells of acute myeloid leukaemia, including megakaryoblasts of megakaryoblastic leukaemia, by hairy cells of hairy cell leukaemia, by centroblasts of centroblastic lymphoma and by plasma cells of plasma cell leukaemia. The enzyme activity was also demonstrated in the mast cells of systemic mastocytosis. It seems that different forms of peroxidase are present in most haemic cells. Fixation and conditions of incubation are probably the determining factors for their demonstration. The method may allow further cytochemical discrimination at the LM level between blast cell populations which are negative for MPO by standard methods.

Use of Karpas HIV cell test to detect antibodies to HIV-2.

Commercial enzyme-linked immunosorbent assays for HIV-1 infection often fail to detect antibodies to HIV-2. A simplified version of the HIV cell test detects antibodies to this new virus and enables typing of HIV infections. Since sera from 3 macaque monkeys infected with simian immunodeficiency virus gave weakly positive reactions, this test holds promise for discovering any further strains of HIV that may exist or evolve.

Cytochemistry of gamma-glutamyltransferase in haemic cells and malignancies.

An improved cytochemical method for the demonstration of gamma-glutamyltransferase is described. The enzyme was demonstrated in almost all normal leucocytes and in platelets. There was markedly reduced activity in most lymphoproliferative disorders. In a single case of Hodgkin's disease, T-lymphocytes showed slight to moderate activity contrasting with the marked activity displayed by Reed-Sternberg cells. The plasma cells of multiple myeloma and plasma cell leukaemia showed activity equal to or stronger than that of their normal counterparts. In acute myeloid leukaemia the positivity varied widely in blast cells, but was consistently increased in monoblasts of acute monoblastic leukaemia. gamma-Glutamyltransferase may serve as a differentiation marker in the study of granulocytic and lymphocytic cell lineages.