Congenital dyserythropoietic anaemia type II: a case study.

A 13-year-old girl with chronic anaemia showed features of congenital dyserythropoietic anaemia (CDA) type II. The main clinical and haematological findings were splenomegaly, a mild microcytic anaemia, and numerous bizarre and binucleate normoblasts in the bone marrow. The acidified serum lysis test (Ham's test) performed with 5 normal sera was positive. The patient's red blood cells showed a markedly increased expression of the i red blood cell antigen.

Simultaneous occurrence of myelodysplastic syndrome and monoclonal B lymphocytes with a different clonal origin.

Bone marrow and peripheral blood from a myelodysplastic syndrome patient with trisomy 13 and monoclonal B lymphocytes (without evidence of systemic lymphoma) were investigated for clonal lymphoid lineage involvement using interphase fluorescence in situ hybridization (FISH) and X-chromosome inactivation assay (HUMARA) on CD19+ and CD34+ sorted cells. Trisomy 13 was detected in 55% of CD34+ cells and in 5.5% of CD19+ cells, the latter being comparable to the negative control specimen. X-chromosome inactivation showed both CD34+ and CD19+ cells to be monoclonal, though their inactivated X-chromosome was different. The results strongly suggested that both populations of CD34+ and CD19+ cells have originated from a different progenitor stem cell.

Chronic myeloid leukemia and interferon-alpha: a study of complete cytogenetic responders.

Achieving a complete cytogenetic response (CCgR) is a major target in the treatment of chronic myeloid leukemia (CML) with interferon-alpha (IFN-alpha), but CCgRs are rare. The mean CCgR rate is 13%, in a range of 5% to 33%. A collaborative study of 9 European Union countries has led to the collection of data on 317 patients who were first seen between 1983 and 1997 and achieved CCgRs with IFN-alpha alone or in combination with hydroxyurea. The median time to first CCgR was 19 months (95% CI, 17-21; range, 3-84 months). At last contact, 212 patients were still alive and in continuous CCgR; 105 patients had lost CCgR, but 53% of them were still alive and in chronic phase. IFN-alpha treatment was discontinued permanently in 23 cases for response loss, in 36 cases for chronic toxicity (15 are still in unmaintained continuous CCgR), and in 8 cases because it was believed that treatment was no longer necessary (7 of these 8 patients are still in unmaintained continuous CCgR). The 10-year survival rate from first CCgR is 72% (95% CI, 62%-82%) and is related to the risk profile. High-risk patients lost CCgR more frequently and more rapidly and none survived more than 10 years. Low-risk patients survived much longer (10-year survival probability 89% for Sokal low risk and 81% for Euro low risk). These data point out that a substantial long-term survival in CCgRs is restricted mainly to low-risk and possibly intermediate-risk patients and occurs significantly less often in high-risk patients.

Trisomy 3q11-q29 is recurrently observed in B-cell non-Hodgkin's lymphomas associated with cold agglutinin syndrome.

Three cases of low-grade B-cell non-Hodgkin's lymphoma associated with cold agglutinin syndrome, cytogenetically characterized by partial trisomy 3, are presented in this report. Our data suggest that the long arm of chromosome 3 might be of particular importance in the pathogenesis of this subgroup of lymphomas.

FISH identifies different types of duplications with 12q13-15 as the commonly involved segment in B-cell lymphoproliferative malignancies characterized by partial trisomy 12.

Clinical, cytogenetic, fluorescence in situ hybridization (FISH), and Southern blot data of 18 patients with different subtypes of B-cell non-Hodgkin's lymphoma, cytogenetically characterized by partial trisomy 12, are presented. These chromosomal changes occurred predominantly in clinically progressive chronic lymphocytic leukemia, mixed cell type, and advanced-stage follicle center cell lymphoma at the time of relapse or transformation into diffuse large cell lymphoma. Partial trisomy 12 consistently included the long arm of chromosome 12, either completely or partially, and resulted from dup(12q) or other rearrangements involving chromosome 12. The duplications were cytogenetically identified as dup(12)(q13q23), dup(12)(q13q22), or dup(12)(q13q15) in follicle center cell lymphoma or t(14;18)-positive diffuse large cell lymphoma; dup(12)(q13q22) or dup(12)(q13q24) in chronic lymphocytic leukemia; and dup(12)(q13q21) in a case of t(14;18)-negative diffuse large cell lymphoma. FISH, using library probes and a panel of YAC probes, mapped along the long arm of chromosome 12, confirmed the cytogenetic results in all cases analyzed except for three cases of t(14;18)-positive follicle center lymphoma or diffuse large cell lymphoma with dup(12q). In these cases, FISH showed similar, possibly identical, duplications, which involved a region more centromeric (12q11-21) than assumed by karyotypic analysis (12q13-22 or 12q13-23) and included alphoid DNA sequences, a combination hitherto unknown. In addition, commonly duplicated regions of chromosome 12 could be defined: 12q11-21, including alphoid DNA sequences for follicle center cell lymphoma or t(14;18)-positive diffuse large cell lymphoma, 12q13-22 for chronic lymphocytic leukemia, and 12p13-q15 for marginal zone cell lymphoma, all of which overlapped in 12q13-15. Whether these regions, especially 12q13-15, may contain genes which are important in malignant transformation or disease progression of B-cell lymphoproliferative malignancies characterized by complete or partial trisomy 12 remains to be determined.

Further characterization of morphologically defined typical and atypical CLL: a clinical, immunophenotypic, cytogenetic and prognostic study on 390 cases.

We analysed a group of 390 patients, diagnosed with chronic lymphocytic leukaemia (CLL). Cases were subclassified as morphologically typical and atypical CLL according to the criteria of the FAB proposal. Typical CLL cases were mostly diagnosed at a low-risk stage (Binet A/Rai 0), required no immediate treatment and expected a long survival; atypical CLL cases mostly presented at a more advanced risk stage (Binet B/Rai I-II), usually required immediate treatment and their survival was shorter. Moreover, clinical staging was of prognostic significance in typical but not in atypical cases. In typical CLL, del(11q) was the most common chromosomal abnormality (21%) whereas in atypical CLL trisomy 12 was found in about 65% of the cases documented with an abnormal karyotype. Although chromosomal abnormalities were associated with a poor survival in typical CLL, they are of no prognostic significance in atypical CLL. Based on these data, we conclude that subtyping CLL by morphology enables the identification of two groups of cases, each characterized by a specific clinical presentation, different cytogenetic abnormalities and prognostic parameters. We speculate that these two groups may represent two related, but different, diseases with different prognostic parameters and a different survival.

Philadelphia chromosome-positive acute myeloid leukemia: cytoimmunologic and cytogenetic features.

BACKGROUND: Little is known about the morphological and clinical features of the minority of acute myeloid leukemias (AML) that carry the t(9;22)(q34;q11) translocation. MATERIALS AND METHODS: Cytologic, cytogenetic and clinical features were studied at diagnosis and during disease evolution in 11 patients presenting with de novo AML. Diagnoses according to the FAB criteria were AML-M2 (3 cases), AML-MO and AML-M4 (2 cases each), AML-M4eos, AML-M5, AML-M6, AML-M7 (1 case each). RESULTS: Immunophenotyping disclosed positivity for CD33 or CD13 and for the CD34 stem cell antigen in all cases tested. Lymphoid-associated markers (LM) were detected in 7/9 patients. In 5 cases the expression of at least 2 LM was seen. In addition, evidence of clonal rearrangement of the immunoglobulin (Ig) and/or T-cell receptor (TCR) genes was documented in 3/4 evaluable cases. The Ph chromosome was found as the sole change in 5/11 cases; the karyotype reverted to normal in 2/4 patients who achieved complete remission. Rearrangement in the M-bcr region was detected by Southern blotting in 2/7 cases. CONCLUSIONS: This infrequent cytogenetic subset of de novo AML appears to be characterized by heterogeneous cytologic features, with frequent expression of lymphoid markers, and by unfavorable prognosis. The combination of clinical, cytologic and cytogenetic studies is important in distinguishing de novo AML with the t(9;22) from chronic myelogenous leukemia blast crisis.

Dicentric (1;15) in myeloid disorders.

We report three cases of myeloid disorders with a dic(1;15)(p11;p11), resulting in trisomy of the long arm of chromosome 1. A review of the literature showed six cases, reported as t(1;15). We suggest that these cases have the same anomaly and should be reappraised as dic(1;15).

Deletions of the long arm of chromosome 7 in myeloid disorders: loss of band 7q32 implies worst prognosis.

Clinical and cytogenetic data were analysed in 54 patients with acute non-lymphocytic leukaemias (ANLL) or MDS (myelodysplastic syndromes) and deletion of the long arm of chromosome 7 (7q-), in order to determine if there is a commonly deleted region in 7q and to establish possible correlations between karyotypic features, such as karyotype pattern, karyotype complexity, associated anomalies, and/or the type of deleted segments, and outcome of patients with these disorders. The median follow-up of our patients was 4 months (range 1-89), as was the median survival. In 30% of the cases there was a history of preceding MDS or previous chemotherapy. Clinical and cytogenetic remission was obtained in 7/36 patients treated with chemotherapy (CT). At the time of 7q- detection, three patients previously treated with CT for ANLL were in clinical remission. 5q- was the most recurrent associated abnormality. Complex karyotypes were observed in 68% of the cases. In univariate analysis, statistical differences in survival were observed according to diagnosis (therapy-related and secondary diseases had a worse prognosis than primary disorders), the chromosomal segments deleted (the loss of band 7q32 was of poor prognostic value), the karyotype complexity (patients with single anomalies did better than patients with complex anomalies) and the response to therapy (patients who achieved complete remission had a better survival probability). In multivariate analysis, the loss of band 7q32 was found to be significantly related to very poor prognosis. This finding suggests that band 7q32 may contain critical genes that should be explored at the molecular level.

BCL3 rearrangement and t(14;19)(q32;q13) in lymphoproliferative disorders.

Translocation t(14;19)(q32;q13) is a rare but recurrent abnormality in chronic lymphocytic leukemia and small cell lymphoma. It has been associated with rearrangements of the BCL3 gene, which is located at the breakpoint on chromosome 19 and is juxtaposed to the immunoglobulin heavy chain locus on chromosome 14 as a result of the translocation. This results in transcriptional up-regulation of the BCL3 gene, which encodes a transcription coactivator, an I-kappa B protein, probably contributing to disease progression. We found, among 4,487 cytogenetic analyses of lymphoproliferative disorders, six cases with a t(14;19)(q32;q13), five of which showed the classical t(14;19)(q32;q13) and one of which showed a three-way translocation t(7;19;14)(q21;q13;q32). The 14;19 translocation never occurred as a single abnormality; additional aberrations included trisomy 12 and several structural abnormalities. The cytogenetic examination was supplemented by molecular analysis using available probes for the BCL3 locus (p alpha 1.4P and p alpha 5B) in 1,150 of the 4,487 patients. Rearrangements of BCL3 could be detected in five cases, all of which had the classical t(14;19). In the case with t(7;19;14), the suspected BCL3 involvement could only be confirmed using long-range restriction mapping, indicating that, with the usually available BCL3 probes, rearrangements of this locus may be missed.

Cytogenetic analysis of B cell chronic lymphoid leukemias classified according to morphologic and immunophenotypic (FAB) criteria.

609 patients with B cell chronic lymphoproliferative disorder were studied with the primary aim of analyzing the cytogenetic profile of B cell chronic lymphocytic leukemias and, if possible, define correlations with FAB classification of these diseases. Morphological and immunological studies were performed according to criteria proposed by the FAB group. A panel of monoclonal antibodies, including at least sIg, CD19, CD5, and FMC7 was used. Interpretations of morphology and cytogenetics were made independently. When applying strict FAB criteria 65% of the cases could be classified. Most of them (44%) were chronic lymphocytic leukemia (CLL). The cases not satisfying strict FAB criteria could be divided into two groups: one closely related to CLL, and here defined as atypical CLL (aCLL) (21%) and another group consisting of patients with leukemic manifestations of B cell non-Hodgkin's lymphoma (LL) (14%). Analyzable metaphases were obtained in 89% of patients. Clonal abnormalities were present in 35% of patients. The most frequent chromosomal changes were abnormalities of chromosome 11q (60 cases), trisomy 12 (46 cases) and structural rearrangements of chromosome 14q (44 cases). Statistical associations with FAB subtypes were found: aCLL and trisomy 12 (P < 0.00001); mantle zone lymphoma (MZL) and t(11;14) (P < 0.00001) and del(6)(q) (P < 0.0001); CLL/mixed cell type and del(6)(q) (P < 0.002); follicular lymphoma and t(14;18) (P < 0.00001); splenic lymphoma with villous lymphocytes and del(7)(q) (P < 0.0004); leukemic lymphoma (LL) with rearrangements in chromosome 9q (P < 0.0001) and trisomy of 3 (P < 0.001). Chronic lymphocytic leukemia was not statistically associated with any specific chromosomal abnormality. However, this subtype showed a high incidence of del(11)(q) and rearrangements of 13q. This study confirms the value of cytogenetic investigation in the diagnosis of these disorders and may provide some new elements for future refinement of the FAB classification in mature B cell lymphocytic disorders.

Isodicentric (X)(q13) in haematological malignancies: presentation of five new cases, application of fluorescence in situ hybridization (FISH) and review of the literature.

Idic(X)(q13) represents a rare but recurrent chromosomal abnormality in haematological malignancies. We present five new cases characterized by this particular aberration and review the literature on this subject. The patients were elderly females with a diagnosis of refractory anaemia (1/5), refractory anaemia with ringed sideroblasts (2/5), chronic myelomonocytic leukaemia (1/5), and Philadelphia chromosome-negative chronic myeloid leukaemia (1/5). Three out of the five patients demonstrated an increased proportion of bone marrow ringed sideroblasts. After a follow-up period of 30-57 months all patients but one are alive. Idic(X)(q13) always occurred as the sole chromosomal abnormality, either in one or in two copies. We confirmed the dicentric nature of the aberration by fluorescence in situ hybridization (FISH) on metaphases as well as interphase nuclei using an X-chromosome-specific alpha-satellite probe, and performed chromosome painting to visualize possible additional chromosomal changes involving the X chromosomes. Our findings and the data of 17 previously published cases indicate that idic(X)(q13): (1) may play a significant pathogenetic role in haematological malignancies affecting exclusively females and deriving predominantly from early progenitor cells; (2) is frequently associated with a pathological iron accumulation; (3) indicates a variable prognosis.

Trisomy 3 is a consistent chromosome change in malignant lymphoproliferative disorders preceded by cold agglutinin disease.

Cold-antibody autoimmune haemolytic anaemia is a rare entity that has been associated with a wide variety of pathological processes, including malignant lymphoproliferative disorders. In this retrospective study we recorded, as far as possible, clinical, haematological, immunological, morphological, pathological, cytogenetic and molecular data on 10 patients with cold agglutinin disease (CAD). Cytogenetic anomalies were found in four cases in which an underlying lymphoma could be evidenced. Trisomy 3 was the only recurrent aberration in our series. It was observed in all patients with abnormal karyotype, either as a complete trisomy or as a partial trisomy of the long arm. The importance of this particular karyotypic aberration in the monitoring of CAD is emphasized.

Mediastinal B-cell lymphoma with sclerosis: clinical features and treatment results in 10 patients.

Mediastinal large-B-cell lymphoma with sclerosis is now considered to be a discrete subtype of lymphoma. It probably originates in the thymus, a T-cell organ. Early publications consider this lymphoma as an aggressive disorder with poor prognosis. We studied retrospectively ten consecutive patients with mediastinal B-cell lymphoma with sclerosis seen in the department of hematology. Nine were women. The median age at diagnosis was 38.3 years (16-60). Dyspnea (experienced by 7 patients), chest pain (5) and cough (10) were the most common clinical features at presentation. Superior vena cava syndrome occurred in three patients. Five had infiltration of the chest wall or of the pulmonary tissue. Four patients were in clinical stage I (all bulky > 10 cm), four in stage IIE, one was in stage IIE and one in stage IV (Ann Arbor classification). All patients were treated with intensive chemotherapy, mostly containing cyclophosphamide, doxorubicin, vincristine or vindesine, bleomycin and prednisone, combined with etoposide or teniposide and methotrexate. Nine patients responded well to chemotherapy (tumor reduction > 75%). One patient progressed. Eight patients received involved field radiotherapy (36-40 Gy) after chemotherapy. The two other patients were treated with intensive chemotherapy (BEAC, BCNU, etoposide, cytarabine, cyclophosphamide), followed by autologous bone marrow transplantation. Two patients died: one patient received autologous bone marrow transplantation in partial remission and relapsed after 6 months; the other patient had progressive disease despite chemotherapy, surgery and radiotherapy. Mean follow-up is 54.6 months (15-118) with 8 patients still remaining in complete remission. In patients with mediastinal B-cell lymphoma, tumour localisation is often limited to the thorax.(ABSTRACT TRUNCATED AT 250 WORDS)

T-lymphoid extramedullary (lymphadenopathic) blast crisis in CML.

T-cells are only rarely involved in chronic myeloid leukemia. We report an unusual case of T-lymphoid extramedullary (lymphadenopathic) blast crisis in a 66-year-old patient with Philadelphia positive chronic myeloid leukemia. The lymph node morphological picture resembled that of a peripheral T-cell lymphoma. Immunohistochemical stainings showed most lymph node blasts to have T-phenotype (positivity for CD-2, CD-3 and CD-5). Flow cytometric phenotyping confirmed T-phenotype, with positivity for CD-7, CD-38 and TdT. Cytogenetic analysis of lymph node cells revealed Philadelphia chromosome with an additional chromosomal abnormality (add(6p)). The BCR-gene rearrangement in the lymph node blasts was identical to that in the bone marrow. This case adds to the growing evidence that CML may be a disorder of the common stem cell from which T-, B- and myeloid progenitors originate.

Cytogenetics of hybrid acute leukemias.

Although the recognition of hybrid acute leukemia (HAL) is still controversial, several reports have described cytogenetic findings in these leukemias over the last 3 years. A distinct chromosomal profile appears to be associated with different immunologic subsets of HAL. The classical t(15;17), and inv(16) as well as abnormalities of the long arm of chromosome 5 and/or 7 are preferentially associated with acute myeloid leukemia (AML) with T-cell features; the t(8;21)(q22;q22), the Ph chromosome, and 11q23 rearrangements are more frequently found in AML with B-cell features; the Ph chromosome, t11q23 and 14q32 breaks without rearrangements of the immunoglobulin heavy chain gene may be associated with acute lymphoblastic leukemia (ALL) with myeloid markers. In addition, some chromosome aberrations may be encountered more frequently in acute leukemia with major phenotype deviations than in unselected cases of acute leukemia: namely the Ph chromosome, 11q23 rearrangements, and +13. These chromosome changes appear to be associated with a low complete remission (CR) rate. An association has been documented in some patients with ALL between the presence of the t(9;22) and a minor myeloid component consisting of 5-15% blast cells with myelomonocytic features, raising the possibility that a diagnosis of bilineal acute leukemia would be more appropriate in such cases. These patients appear to have a severe outcome with significantly lower CR rate than similar cases of Ph-positive ALL without a minor myeloid component.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between nuclear morphology and DNA content in large cell lymphomas.

The relationship between DNA content and nuclear morphology in large cell lymphomas (LCLs) was investigated on lymph node imprints. Mean maximum nuclear diameter (mean MND), nuclear shape and chromatin pattern were evaluated microscopically on May-Grünwald-Giemsa-stained slides. DNA content and nuclear area were measured on Feulgen-stained slides by image analysis. Twelve of the 24 cases were DNA diploid and 12 tetraploid. The DNA diploid cases were characterized by medium large (mean MND 11.2-13.7 microns), round nuclei with a fine chromatin pattern. The DNA tetraploid cases had significantly (P < .01) larger (mean MND 13.0-19.1 microns) nuclei and a higher frequency of coarse chromatin pattern, nuclear irregularities and multilobation. One of the researchers, unaware of the DNA index, could predict the ploidy level in 80% of cases on morphology. The linear coefficient of correlation between mean MND and mean nuclear area was 0.84. We also found that nuclear area increased as the cell moved through the cell cycle. DNA content, related to ploidy and position in the cell cycle, is an important explanation for the variable nuclear morphology in LCLs.