Trisomy 12-associated, t(11;14)-negative mature B-cell leukemia with gene expression profile resembling mantle cell lymphoma

Trisomy 12 can be seen in many different lymphoid neoplasms. However, many or most mature B-cell leukemias associated with isolated trisomy 12 are reported in the literature as chronic lymphocytic leukemia (CLL) or 'atypical CLL'. This study reports a case of a mature B-cell leukemia, morphologically and immunophenotypically similar to cases previously published as atypical CLL, in which cytogenetic evaluation revealed an isolated clonal trisomy 12 but no evidence of the mantle cell lymphoma-associated t(11;14)(q13;q32). Further analysis confirmed absence of cyclin-D1 expression. Subsequent lymph node biopsy revealed evidence of large cell transformation of the underlying chronic lymphoproliferative disorder. Gene expression profiling of the initial peripheral blood sample using a cDNA micro-array of ∼10,000 expressed genes revealed a close resemblance between the reported case and 2 cases of known mantle cell lymphoma. When further compared to 7 known 'typical' CLL cases, the reported case was classified as mantle cell lymphoma by hierarchical cluster analysis. The case reported here raises interesting questions regarding the nature of cases reported previously as trisomy 12-associated CLL and reinforces the fact that other leukemic lymphoproliferative disorders should be included in the differential diagnosis of such cases. Further study is indicated to elucidate the nature and diversity of disorders previously reported as trisomy 12-associated chronic lymphocytic leukemia.     

Immunophenotyping of subtypes of B-chronic (mature) lymphoid leukemia. A study of 242 cases.

BACKGROUND. The French-American-British group's proposal for the classification of chronic lymphoid leukemias is unique at this time. Testing, expanding, and adding to the theory by immunophenotyping will help to additionally characterize this group of diseases. METHODS. Peripheral blood samples from 242 patients with chronic lymphoid leukemias were analyzed for immunologic evaluation of the following subtypes: typical chronic lymphocytic leukemia (CLL), 189; CLL with pleomorphic lymphocytes (CLL-pleo), 19; CLL of mixed cell type (CLL/PL), 20; prolymphocytic leukemia (PLL), 22; hairy cell leukemia (HCL), 10; HCL-variant, 1; and splenic lymphoma with villous lymphocytes, 1. RESULTS. The phenotype of CLL and CLL-pleo was weak surface immunoglobulin (SIg) with positive results of mouse rosettes (MR+), CD5+, and CD22-. Of PLL and HCL, it was strong SIg, MR-, CD5-, and CD22+. By analyzing the four markers and accepting the relevant results of two or more as sufficient for diagnosis, all cases (100%) of CLL, CLL-pleo, PLL, and HCL were diagnosed. CLL/PL showed the phenotype of CLL in 66.67% and of PLL in 33.33% of patients. The frequency of cases with weak fluorescence in decreasing order was CLL, CLL-pleo, CLL/PL, and PLL and HCL. The same sequence applied to the mean percentage of mouse rosette-forming cells and CD5 cells, but the sequence was reversed for CD22 cells. CONCLUSIONS. SIg intensity, MR, CD5, and CD22 constitute the minimum number of immune markers for the differential diagnosis of the subtypes of chronic lymphoid leukemia. The frequency of the four markers among the subtypes suggested that CLL and CLL-pleo have identical phenotypes and that the five subtypes follow a continuous range of B-cell differentiation from early mature (CLL and CLL-pleo) to late mature pre-plasma cell stages (PLL followed by HCL), with CLL/PL of intermediate maturity.     

Detection of 14q32.33 translocation and t(11;14) in interphase nuclei of chronic B-cell leukemia/lymphomas by in situ hybridization

Abnormalities of chromosome 14 involving band q32.33 are among the most commonly observed cytogenetic alterations in B-cell malignancies. To assess the incidence and pathogenetic implications of 14q32.33 translocation in chronic B-cell leukemia/lymphomas, we performed fluorescence in situ hybridization (FISH) analysis with variable region (V_H) and gamma constant region (Cγ) gene probes in 37 patients with these disorders. Chromosome 14q32.33 translocation was detected in 2 of 18 patients with chronic lymphocytic leukemia (CLL), 1 of 2 with CLL of mixed cell types (CLL/PL), 1 of 2 with pro-lymphocytic leukemia (PLL), 5 of 6 with leukemic mantle-cell lymphoma (MCL), 2 of 7 with splenic B-cell leukemia/lymphoma of possible marginal zone origin (SBLL) and 2 with leukemic follicular lymphoma (FL). To further characterize 14q32.33 translocations in these patients, we developed a new procedure using double-color FISH with PRAD1, BCL2, V_H and Cγ gene probes. Chromosome t(11;14) was detected in 1 patient with CLL/PL, 1 with PLL and 5 with MCL. Chromosome t(14;18) was detected in 2 patients with FL. In a PLL patient with t(11;14), the cosmid CPP29 containing the PRAD1 gene and its 5′-flanking region split and co-localized with both Cγ and V_H gene probes, thus spanning the breakpoint. In CLL and SBLL patients, donor chromosomes were other than chromosomes 2, 11, 18 and 19, suggesting the involvement of a novel oncogene(s) in the pathogenesis of these diseases. Interphase FISH rapidly detected 14q32.33 translocation, t(11;14) and t(14;18) in B-cell malignancies with low mitotic activity at the single-cell level, facilitating the correlation of the molecular features of these translocations with clinical characteristics. Int. J. Cancer 72:31–38, 1997. © 1997 Wiley-Liss Inc.     

Mosaicism of trisomy 12 in chronic lymphocytic leukemia detected by non-radioactive in situ hybridization.

Cytogenetic analysis using banding techniques of B-chronic lymphocytic leukemia (CLL) is hampered by the difficult in vitro proliferation of these tumor cells. For detection of specific cytogenetic aberrations these problems can be overcome with non-radioactive in situ hybridization (ISH). ISH may especially be applied for the detection of trisomy 12, which is the most frequent cytogenetic aberration in CLL. Sixty-seven patients with CLL, four normal controls and one lymphoblastoid B-cell line with a trisomy 12 were studied using a chromosome 12 specific probe. To determine the hybridization properties of the CLL cells, all samples were also hybridized with probes specific for chromosomes 1 and 8. All leukemias were analyzed by immunocytochemistry to determine the proportion of tumor cells. Eight cases (11%) showed a trisomy 12. After correction for the number of tumor cells, it was demonstrated that in almost all cases (7 out of 8), the aberration was present in a proportion of the tumor cells (between 30 and 72%). Except for one patient this mosaicism persisted with long-term follow-up. We conclude that the in vivo incidence of trisomy 12 in CLL is approximately 11%, and that trisomy 12 occurs in most instances in only a subpopulation of the leukemic cells. Both findings suggest that trisomy 12 in CLL is a late event.     

Subtype Distribution,Clinical Features,and Survival in B-cell Chronic Lymphoproliferative Disorders in China: A Review of 1592 Cases

BackgroundB-cell chronic lymphoproliferative disorders (B-CLPDs) are characterized by the sustained accumulation of monoclonal B cells. Limited studies have systematically described the clinical features and outcomes of the whole patient group, especially in Eastern populations.Patients and MethodsA total of 1592 patients with newly diagnosed B-CLPD were enrolled. Chronic lymphocytic leukemia (CLL) accounted for 39%, and Waldenström macroglobulinemia (WM), leukemic marginal zone lymphoma, follicular lymphoma (FL), and mantle cell lymphoma (MCL) constituted 13%, 13%, 9%, and 8% of cases, respectively.ResultsThe median age at diagnosis was 58 years, and the male/female ratio was 1.8:1. The 17p and 11q deletions were most common in MCL (36% and 17%, respectively), and 13q deletion and trisomy 12 were most frequent in CLL (35% and 21%, respectively). Patients with leukemic MCL had significantly worse survival than that of patients with other disease entities, with a 3-year overall survival (OS) of 58%, followed by 68.2% for WM/lymphoplasmacytic lymphoma. Those with CLL, leukemic marginal zone lymphoma, and FL had relatively favorable outcomes, with a 5-year OS > 80%. The survival of patients with B-CLPDs has improved over time with the emergence of novel drugs (3-year OS improvement from 82.1% to 92.2%). The improvement in survival mainly resulted from improvement among patients with MCL, WM/lymphoplasmacytic lymphoma, and FL. On multivariate analysis, only hemoglobin, lactate dehydrogenase, and 17p deletion were independently associated with survival (hazard ratio, 1.6, 2.0, and 3.1, respectively).ConclusionsComprehensive analysis of the clinical characteristics, immunophenotypic profiles, and cytogenetic features can be helpful in the differential diagnosis, especially for patients without a non–bone marrow biopsy specimen available. Universal prognostic factors could help with the early detection of high-risk patients and stratification for risk-adapted therapy.     

Surface Phenotype and Adhesion Activity of B-Cell Chronic Lymphoid Leukemias

Surface phenotypes and adhesion activity to human umbilical vein endothelial cells (HUVECs) were studied using leukemic cells from 12 Japanese patients with B-cell chronic lymphoid leukemias including 7 with chronic lymphocytic leukemia (CLL), 1 with prolymphocytic leukemia (PLL), 2 with hairy cell leukemia (HCL) and 2 with HCL variant (HCL-V). CD 13 and CD23 were found to be characteristically positive in CLL, whereas they were not expressed in non-CLL cases except for positivity of CD23 in two such cases. Except for CDl lb, all other leukocyte integrins examined (CDlla, CD11c and CD18) and their ligand (CD54) were highly expressed in non-CLL cases. Adhesion activity of leukemic cells to HUVECs after co-culture with HUVECs was well correlated with the expression of CD11b lb, CD18 and CD54, but showed no predilection for any leukemia subtype. Positivity for CD5, CD21, CD23 and CD13 changed after the co-culture with HUVEC. These results suggest that adhesion activity after co-culture does not correlate with the leukemia subtype and that endothelial cells activate or differentiate leukemic cells.     

T- and B-lymphocytes in lymphoproliferative diseases]

T-cell and B-cell markers and the response of lymphocytes to phytohemagglutinin (PHA) have been studied in 62 patients with chronic lymphocytic leukemia (CLL), in 3 patients with well-differentiated lymphocytic lymphoma and in 1 patient with lymphosarcoma. To investigate the functional integrity of T-lymphocytes in 10 patients with CLL the response of the T-cell-rich population to PHA was determined. The leukemic lymphocytes in all the cases but one had B-cell characteristic. The T-cell-rich population would show a normal response to PHA.     

慢性淋巴细胞白血病的分子遗传学特点

目的 了解慢性淋巴细胞白血病（CLL）的分子遗传学特性。方法运用间期荧光原位杂交（FISH）技术对60例初发的B细胞CLL（B-CLL）患者进行12号染色体3体（＋12）、del（13q14）和del（17p13）检测。结果60例患者中,41例（68．3％）至少有一种分子遗传学异常,2例（3．3％）具有2种染色体异常。12例（20．0％）有＋12异常,其畸变细胞率在4．0％-34．0％之间;24例（40．0％）有del（13q14）异常,其畸变细胞率在22．0％～93．0％之间,其中3例有2条染色体del（13q14）异常;7例（11．7％）有del（17p13）异常,其畸变细胞率在6．0％～68．0％之间。不同Binet分期中,3种分子遗传学异常差异无统计学意义。结论FISH是一种在分析CLL染色体数目和结构异常方面较为快速、准确和敏感的方法,可为CLL的研究提供较为准确的分子遗传学信息。     

Monoclonal antibody Ki-67 identifies B and T cells in cycle in chronic lymphocytic leukemia: correlation with disease activity.

Ki-67 is a monoclonal antibody that recognises a nuclear antigen expressed during most phases of the cell cycle. We have analysed, by immunocytochemistry, the frequency, morphology, and clinical significance of Ki-67+ cells in 108 patients with B-cell chronic lymphocytic leukemia (CLL). Because in normal peripheral blood Ki-67+ cells are mainly T lymphocytes, we have also investigated, by double immunoenzymatic staining, the proportion of Ki-67+ T cells (Ki-67/CD3+) in CLL. Four groups of patients were identified: (i) 47 with stage A, (ii) 32 with stages B + C, (iii) 24 with greater than 10% of circulating prolymphocytes (CLL/PL) and (iv) five with Richter's syndrome. Within stage A CLL, two groups were considered: A' (Hb greater than or equal to 12 g/dl and lymphocytes less than 30 + 10(9)/l) and A" (Hb less than 12 g/dl or lymphocytes greater than or equal to 30 x 10(9)/l). The percentage and absolute number of Ki-67+ leukemic cells was found to increase with the stage of the disease and correlate with the proportion of prolymphocytes. On the other hand, the proportion of Ki-67+ T cells (CD3+) was significantly higher in patients with CLL stage A' (29.3 +/- 4.5), which includes patients with long-standing, stable disease, than in CLL stage A" (9.5 +/- 3.3), B + C (7.1 +/- 4.6), and CLL/PL (6.4 +/- 2.8). Ki-67 seems to identify patients with more aggressive forms of CLL, such as CLL/mu 2PL with more than 10% Ki-67+ cells (25% of the cases) and Richter's syndrome, in which all the large lymphoma cells are Ki-67+. Long-term follow-up will establish whether Ki-67 is a good prognostic marker and can predict disease outcome.     

Atypical lymphocytes in B-cell chronic lymphocytic leukemia and trisomy 12 studied by conventional staining combined with fluorescence in situ hybridization

Trisomy 12 is one of the most frequent chromosomal abnormalities in B-cell chronic lymphocytic leukemia (CLL), and is predominantly found in CLL with atypical morphology (aCLL). It has been suggested that the atypical morphology might be a feature of the abnormal trisomy 12 clone, but so far it has been difficult to allocate chromosomal aberrations to individual leukemic cells identified by cytomorphology. We therefore wanted to use our MGG/FISH method, which combines fluorescence in situ hybridization (FISH) and standard cytomorphology, to study if the trisomy 12 clone in CLL was restricted to lymphocytes with atypical morphology. Peripheral blood specimens of four patients with aCLL were studied using a DNA probe against the pericentromeric region of chromosome 12. Trisomy 12 was found in 10-34 % of the lymphocytes. In three patients, the proportion of atypical and typical lymphocytes with trisomy 12 was quite comparable, and so was the percentage of atypical cells with lymphoplasmacytoid morphology and those with cleaved nucleus showing trisomy 12. Only one patient differed, since we found an overrepresentation of trisomy 12 among the atypical lymphocytes. However, this could be fully explained by the diluting effect of contaminating T-cells after chemotherapy. The results of the present study show that despite the strong association of trisomy 12 and atypical morphology in CLL, this chromosomal abnormality is not confined to lymphocytes with atypical morphology, but is also found in typical CLL cells. This supports that both cell types have the same clonal origin and that different cell morphology cannot be explained alone by the acquisition of an additional chromosome 12.     

Cellular distribution of a B-cell specific surface antigen (gp54) detected by a monoclonal antibody (anti-BL4)

A monoclonal antibody (anti-BL4) recognizing a previously characterized Mr 54,000 glycoprotein (gp54) was developed by immunizing BALB/c mice with cells from a precursor B-cell line (Josh-7). In normal individuals, this antigenic molecule was present on tonsillar B-cells (60-80%) and on a fraction of peripheral blood B-cells (5-25%). BL4 (gp54) expression was investigated in 186 patients with a variety of hematological malignancies using indirect immunofluorescence and flow cytometric analysis. Twenty-six of 37 cases of B-cell chronic lymphocytic leukemia (CLL) and 18 of 33 cases of B-cell non-Hodgkin's lymphoma were BL4 positive. Surface expression of BL4 on reactive cases of CLL and non-Hodgkin's lymphoma was brighter than those of B1, B2, and B4, BL4 positive CLL cases expressed a higher proportion of mouse rosette forming cells and Leu-1 positive cells than the BL4 negative subgroup and were not associated with elevated serum immunoglobulin levels. Four of 7 BL4 negative CLL cases were associated with increased serum levels of immunoglobulin M. Lymphoblasts from 14 of 14 cases of non-T acute lymphoblastic leukemia and 3 of 3 pre-B lymphoid blast crisis of chronic myeloid leukemia were BL4 negative. Neoplastic cells from 2 of 3 cases of Waldenstrom's macroglobulinemia and 4 of 7 cases of hairy cell leukemia were BL4 reactive. None of 7 cases of multiple myeloma and plasma cell leukemia were BL4 positive. All 11 T acute lymphoblastic leukemia cases, 6 other T-cell malignancies, 5 cases of Hodgkin's disease, 51 cases of acute nonlymphocytic leukemia, and 9 cases of chronic myeloid leukemia in chronic phase thus far studied were BL4 negative. An in vitro induction experiment using phorbol ester on a case of B-CLL demonstrated disappearance of BL4 accompanied with further B-cell differentiation. Our study further substantiates the previous finding that gp54 is a differentiation antigen restricted to the B-cell lineage and expressed during the intermediate stage of B-cell ontogeny.     

The clinical and diagnostic relevance of CD23 expression in the chronic lymphoproliferative disease

BACKGROUND: CD23 antigen is a cell surface protein considered important in the differentiation of chronic lymphocytic leukemia (CLL) from other lymphoid leukemias. METHODS: To better clarify CD23 role as a diagnostic tool, the authors retrospectively evaluated clinical and laboratory features of 372 patients who were referred to M.D. Anderson Cancer Center with a diagnosis of CLL or B-cell chronic lymphoproliferative disease. RESULTS: Most of the patients (91%) were CD19+/CD5+. Only 6% of these CD19+/CD5+ patients were CD23-. Overall, CD23- patients had the worse prognostic features compared with CD23+ cases, including anemia (P = 0.03), massive splenomegaly (P = 0.000), high lactate dehydrogenase (P = 0.007), high beta2-microglobulin (P = 0.006), older age (P = 0.001), and male gender (P = 0.02). Surface immunoglobulin expression was moderate/strong in 19 (82%) patients, and FMC-7 was positive in 22 (96%) patients. None of the 13 patients tested for CD10 expressed the antigen. Based on morphology, of the CD23, 16 (70%) were diagnosed with mantle cell leukemia (MCL) was diagnosed in 16 (70%) CD23- patients, 3 (13%) with splenic marginal-zone leukemia, 3 (13%) with prolymphocytic leukemia (PLL) or PLL/CLL, and 1 (4%) with CLL. No cyclin D1 protein expression was noted by Western blot analysis in the one case that showed typical CLL morphology, and this patient did not require therapy. On the whole, the survival rate of CD23- patients was significantly worse than that of patients with CD23+. In contrast, 15 of 32 (49%) CD19+/CD5- patients were CD23-. CD23 negativity in this group was not associated with distinct clinical features or outcome. Eleven (73%) of these patients were classified as having splenic marginal-zone lymphoma and 4 as having follicular lymphoma. CONCLUSIONS: These data indicate that CD23 negativity is rare in typical B-cell CLL, and CD23 negativity in patients with CD19+/CD5+ is suggestive of mantle cell leukemia a more aggressive disease with poor response to conventional therapy in which newer chemotherapy regimens such as hyper-CVAD may be more effective.     

慢性淋巴细胞白血病的Richter’s征转化

目的:探讨Richter’s征(RS)的临床特征、预测指标、治疗和预后因素。方法:报告2例慢性淋巴细胞白血病(CLL)向RS转化并结合文献进行复习。结果:患者表现为进行性淋巴结肿大,伴乏力、盗汗,经血细胞形态学、免疫分型确定存在CLL克隆,淋巴结病理1例示弥漫性大B细胞淋巴瘤、另1例为小B细胞淋巴瘤向大细胞淋巴瘤转化。结论:RS为少见疾病,预后较差,诊断时需有病理证实CLL向侵袭性淋巴瘤转化,治疗应采用含利妥昔单抗的多药联合化疗,年轻患者如对初始治疗反应较好且有合适供体应进行异基因移植。     

CLLU1 expression distinguishes chronic lymphocytic leukemia from other mature B-cell neoplasms

The distinction of CLL from other mature B-cell neoplasms, especially from leukemic forms of mantle cell lymphoma or splenic marginal zone lymphoma, can be difficult but has important prognostic and therapeutic implications. We measured CLLU1 (CLL upregulated gene1) mRNA by qPCR and found a highly significant difference between CLL and other lymphoid neoplasms (AUC 0.96, 95%CI 0.93-0.99). Based on our cut-off values we can predict CLL and other mature B-cell neoplasms with high probability (PPV 99% and 94%). Analysis of CLLU1 expression is a rapid and reliable tool that may facilitate the diagnosis of mature B-cell neoplasms especially in inconclusive cases.     

荧光原位杂交技术检测慢性B-CLL第12号染色体三体畸变

目的 检测２０例慢性Ｂ淋巴细胞白血病（Ｂ－ＣＬＬ）中第１２号染色体三体畸变。方法 运用荧光原位杂交技术（ＦＩＳＨ）。结果 ６例（３０％）患者有１２号染色体三体（＋１２）,其中５例经染色体核型分析发现的＋１２均得到证实。在１例Ｂ－ＣＬＬ中,染色体分析未发现任何分裂相,而ＦＩＳＨ检测发现了＋１２克隆。在另１例Ｂ－ＣＬＬ中,ＦＩＳＨ不仅证实了染色体核型检查发现的存在于扁桃体的＋１２,同时还发现了染色体分     

Molecular allelokaryotyping of early-stage, untreated chronic lymphocytic leukemia

BACKGROUND: To the authors' knowledge, genetic abnormalities in early-stage chronic lymphocytic leukemia (CLL) have not been examined fully. Single nucleotide polymorphism (SNP) genomic array (SNP-chip) is a new tool that can detect copy number changes and uniparental disomy (UPD) over the entire genome with very high resolution. METHODS: The authors performed SNP-chip analysis on 56 samples from patients with early-stage, untreated CLL. To validate the SNP-chip data, fluorescence in situ hybridization (FISH) analysis was performed at selected sites. Expression levels of ZAP-70 and the mutational status of immunoglobulin heavy-chain gene also were examined. RESULTS: SNP-chip analysis easily detected nearly all changes that were identified by FISH, including trisomy 12, deletion of TP53 (17p13), deletion of ATM (11q22), and deletion of 13q14. Only 10 of 56 CLL samples (18%) had no genomic abnormalities. Excluding the 4 common abnormalities mentioned above, 25 CLL samples (45%) had a total of 45 copy number changes detected by SNP-chip analysis. Four samples had 6q deletion at 6q21 that involved the AIM1 gene. UPD was detected in 4 samples; 2 samples involved whole chromosome 13 resulting in homozygous deletion of micro-RNA-15a (miR-15a)/miR-16-1. CLL samples with deletion of 13q14 and trisomy 12 were mutually exclusive. CONCLUSIONS: Genetic abnormalities, including whole chromosome 13 UPD, are very common events in early-stage CLL. SNP-chip analysis can detect small genetic abnormalities in CLL and may be able to support or even supplant FISH and cytogenetics.     

荧光原位杂交技术检测慢性淋巴细胞白血病p53基因的缺失

背景与目的：p53基闪的点突变是慢性淋巴细胞白血病（CLL）患者较频发的分子事件，B—CLL细胞有丝分裂活性低，且对有分丝裂原反应差，因而常规细胞遗传学方法不一定能真实地反映核型状况，本文采用FISH技术进行间期细胞核分析CLL中p53基因缺失情况，探讨其与CLL疾病进展的关系。方法：运用荧光素Spectrum—Red直接标记的p53单一序列DNA为探针的荧光原位杂交（FISH）技术对83例初发的B细胞CLL（B—CLL）患者的间期细胞进行p53基因的检测结果：83例B—CLL中10例（12．0％）有p53基因缺失，其阳性细胞率在6．0％～80．0％之间。p53基因缺失在不同Binet分期中分别为A期6／63（9．5％）、B期2／15（13．3％）、C期2／5（40．0％），A期与C期p53基因缺失阳性率比较有湿著性差异（P〈0．01）。结论：p53基因缺失与部分CLL的发生发展密切相关，FISH是一种在分析CLL p53基因缺失异常方面较为快速、准确和敏感的方法。     

Analysis of DNA aneuploidy as a tumor marker

Cellular DNA content was determined by flow cytometry for 185 adult cases with various types of leukemia. DNA aneuploidies (DA) were detected in 50 of 185 patients (27%). Incidences of DA in subtypes of leukemia were as follows: 26% in acute lymphocytic leukemia (ALL), 17% in acute non-lymphocytic leukemia (ANLL), 28% in chronic myelocytic leukemia in blastic crisis (CML-BC), 6% in chronic lymphocytic leukemia and 55% in adult T-cell leukemia (ATL). Among subtypes, the incidence in ATL was significantly high (p less than 0.01), which suggested the special entity of this disease. No differences of incidence were found between male and female patients. DNA index (DI) ranged between 0.913 and 2.042, and the mean value was 1.144. Hypodiploid clones were detected in two cases (4%) and the rest of cases all showed hyperdiploid clones. Also only in two cases (4%), two abnormal DNA stemlines were detected, which suggested the low incidence of heterogeneous population in leukemic cells. In a few cases , whom we were able to follow up the clinical course, we could detect the early relapse in advance to morphological diagnosis and identify the "recruitment" of leukemic cells with the use of DA as a tumor marker. With regards to the prognosis of patients, DA was found to be useful as an unfavorable prognostic factor in ANLL (p less than 0.01) as well as in CML-BC (0.05). These data showed a clinical usefulness of flow cytometric analysis of DNA aneuploidy as a tumor marker. Moreover, based on our results, the usefulness of flow cytometric analysis of DA was discussed, with reference to current studies on leukemias and solid tumors.     

白血病和淋巴瘤常见染色体融合基因的检测分析

 目的　应用多重巢式RT PCR法 ,结合染色体核型检查分析 75例不同类型白血病观察其治疗和预后。方法　收集 4 1例AML、8例CML、1例CLL、1例PLL、6例MDS、8例NHL、4例MM、5例ALL及 1例嗜酸细胞性白血病患者骨髓标本进行RT PCR及染色体核型检查。结果　 (1)RT PCR示 :75例白血病中 30例阳性条带 ,4 5例无阳性条带。其中M 3:PML/RARA的阳性率为 5 0 % ,PLZF/RARA 8.3% ,(包括 3/ 12例门诊复查病例 ,2 / 12例为同一患者 )。M 4 :CBFB/MYH114 4 .4 % ,MLLex7/MLLex2 11.1%。M5 :MLL/AF6 33.3%。CML :BCR/ABL 87.5 %。MDS :MLLex7/AF4 16 .7% ,EVI1(3q2 6 ) 16 .7%。NHL :EVI1(3q2 6 ) 12 .5 %。MM :HOX115 0 .0 %。ALL :BCR/ABL 2 0 .0 %。 (2 )染色体核型检查多数与融合基因PCR结果相符 ,2例染色体核型检查正常而融合基因检测有阳性条带。结论　 (1)某些融...     

Diffuse large cell lymphoma occurring in a patient with Waldenstr?m's macroglobulinemia. Evidence for the two different clones in Richter's syndrome

The authors report a 60-year-old man with Richter's syndrome, or diffuse large cell lymphoma (DLCL) occurring in a patient with either chronic lymphocytic leukemia (CLL) or Waldenstr?m's macroglobulinemia (WM). Surface marker analysis revealed that the WM showed mu kappa surface immunoglobulin (Ig) chains, and that the DLCL showed mu lambda Ig chains. Flow cytometric DNA analysis demonstrated DNA content differences between WM and DLCL, the former diploid and the latter aneuploid. The current study suggests that Richter's syndrome derives from two independent B-cell malignancies.