The predictive value of initial cytogenetic studies in 148 adults with acute nonlymphocytic leukemia: a 12-year study (1970-1982)

When leukemic blood or marrow specimens from 148 adults with newly diagnosed acute nonlymphocytic leukemia (ANLL) were studied with chromosome banding techniques, 79 were found to have clonal abnormalities. Among 130 treated patients, the 53 with initially normal karyotypes had a significantly longer survival rate than the 16 in whom no normal metaphases were observed (p = 0.02). The 55 patients with both normal and abnormal metaphase cells had an intermediate survival. Once a complete remission had been attained, however, there was no significant difference in median survival between those patients with entirely normal karyotypes and those with abnormal karyotypes. Among the various FAB morphologic subsets of ANLL, the differences in complete remission rate and overall survival between the various cytogenetic subsets were greatest in acute myelogenous leukemia (AML, M1 + M2). The presence of an abnormal clone was a more important predictor of clinical outcome (p = 0.02) than the presence of normal stem cell clones. Aneuploidy alone (hyperdiploidy or hypodiploidy) was not of predictive value, indicating that the use of banding techniques to identify structural rearrangements in pseudodiploid cells was essential. Clonal chromosomal abnormalities were nonrandom and acquired, and specific abnormalities were closely associated with specific clinical-pathologic subsets of ANLL. All 13 patients with acute promyelocytic leukemia and adequate cytogenetic specimens had t(15;17); this translocation was not found in any other subset of ANLL. Six patients with AML (M2) had t(8;21) or a variant of this rearrangement. Seven patients had inv(16)(p13q22) associated with acute myelomonocytic leukemia (AMMoL, M4) and abnormal marrow eosinophils. Two patients had ins(3;3) and thrombocytosis. Four patients had a translocation involving 11q, but none of these had acute monocytic leukemia (AMoL, M5); no patient had del(11q).     

MLL-CBP fusion transcript in a therapy-related acute myeloid leukemia with the t(11;16)(q23;p13) which developed in an acute lymphoblastic leukemia patient with Fanconi anemia

We describe a boy with Fanconi anemia (FA) who developed acute lymphoblastic leukemia (ALL) (FAB-LI) followed by acute myeloid leukemia (AML) (FAB-M5) at relapse. The patient was diagnosed with early pre-B-cell ALL without preceding aplastic anemia and was treated with ALL-oriented chemotherapy which included doxorubicin (a total dose of 140 mg/m(2) administered), which is a topoisomerase II inhibitor. Complete remission was obtained, but after 38 weeks AML developed. The karyotype of ALL cells at diagnosis showed 46,XY, and that of AML cells at relapse was 46,XY, t(11;16)(q23;p13). An MLL gene rearrangement and MLL-CBP chimeric mRNA were found in AML, but not in ALL. A diagnosis of FA was confirmed by an increased number of chromosomal breaks and rearrangements in peripheral blood lymphocytes cultured with mitogen in the presence of mitomycin C. We conclude that this FA patient developed ALL followed by a therapy-related t(11;16)-AML resulting in an MLL-CBP fusion. Further examination of such patients would shed light on leukemogenesis in FA patients. Genes Chromosomes Cancer 27:264-269, 2000.     

Secondary acute myeloid leukemia in children treated for acute lymphoid leukemia

We studied the risk of the development of acute myeloid leukemia (AML) during initial remission in 733 consecutive children with acute lymphoid leukemia (ALL) who were treated with intensive chemotherapy. This complication was identified according to standard morphologic and cytochemical criteria in 13 patients 1.2 to 6 years (median, 3.0) after the diagnosis of ALL. At three years of follow-up, the cumulative risk of secondary AML during the first bone marrow remission was 1.6 percent (95 percent confidence limits, 0.7 and 3.5 percent); at six years, it was 4.7 percent (2 and 10 percent). The development of secondary AML was much more likely among patients with a T-cell than a non-T-cell immunophenotype (cumulative risk, 19.1 percent [6 and 47 percent] at six years). Sequential cytogenetic studies in 10 patients revealed entirely different karyotypes in 9, suggesting the induction of a second neoplasm. In eight of these patients, the blast cells had abnormalities of the 11q23 chromosomal region, which has been associated with malignant transformation of a pluripotential stem cell. There was no evidence of loss of DNA from chromosome 5 or 7, a karyotypic change commonly observed in cases of AML secondary to treatment with alkylating agents, irradiation, or both. We conclude that there is a substantial risk of AML in patients who receive intensive treatment for ALL, especially in those with a T-cell immunophenotype, and that 11q23 chromosomal abnormalities may be important in the pathogenesis of this complication.     

急性白血病患者MSC的免疫原性及抑制异体T淋巴细胞增殖功能和机制研究

目的：研究分析急性白血病患者的骨髓间充质细胞（MSC）对异体T淋巴细胞增殖功能的抑制作用及其作用机制。 方法：选取30例确诊急性髓细胞白血病（AML组）患者及30例急性淋巴细胞白血病（ALL组）,同期健康研究对象30例（健康组）,分离培养三组研究对象的MSC细胞,测定三组MSC细胞培养上清液中细胞因子的表达水平,采用Transwell培养检测三组MSC对T淋巴细胞增殖的抑制作用,采用混合淋巴细胞方法检测MSC对异体T淋巴细胞增殖的抑制作用。 结果： AML组上清液中的TGF-β1、HGF水平显著低于ALL组和健康组（P<0.05）,ALL组的IL-11水平显著高于AML组、健康组（P<0.05）;三组间MSC上清液中IL-6的水平差异无统计学意义（P>0.05）。AML、ALL、健康组的MSC细胞分泌的细胞因子对T淋巴细胞增殖仍然具有显著的抑制作用,接触性共培养和不直接接触的情况下对T淋巴细胞增殖的抑制作用差异无统计学意义（P>0.05）;在加入MSC后,三组的T细胞增殖作用受到显著的抑制,在加入抗TGF-β1抗体、抗HGF抗体后T细胞增殖抑制作用较单纯加入MSC时明显降低（P<0.05）。 结论：急性白血病患者的MSC对异体T淋巴细胞增殖具有抑制作用,其作用原理可能与细胞因子的分泌有关。     

Chromosome abnormalities in acute lymphoblastic leukemia

Less information is available on the cytogenetic abnormalities in marrow cells of patients with acute lymphoblastic leukemia (ALL) than on abnormalities in acute nonlymphocytic leukemia (ANLL); nonetheless, some patterns of karyotypic change in ALL are evident. Even with banding, about 50% of patients appear to have a normal karyotype. The modal chromosome number tends to be higher in ALL than in ANLL. Every patient with B-cell ALL has had an abnormality of one chromosome No. 14 that involved the translocation of material to the end of the long arm. Among seven reported cases, the translocation was from 8q in three patients and 11q in one. Cells with a haploid or near-haploid (24–35) chromosome number have been reported in five patients with ALL and in four patients in a lymphoid blast crisis of chronic myelogenous leukemia. The karyotype in the four ALL patients whose cells were analyzed with banding was remarkably consistent. All patients had the haploid number, usually with both sex chromosomes, plus an additional No. 10, 18, and 21. Evolution of the karyotype, which occurs in the leukemic cells of about 50% of patients, involves cells of patients who had an initially normal or an initially abnormal karyotype. The evidence regarding a correlation between the presence of an abnormal clone prior to treatment and response to treatment is contradictory at present. Some chromosome abnormalities, such as the presence of a Philadelphia (Ph¹) chromosome, a 14q+ chromosome, or a haploid clone, are associated with a relatively short survival.     

Phenotypic and functional comparison of mesenchymal stem cells derived from the bone marrow of normal adults and patients with hematologic malignant diseases

Mesenchymal stem cells (MSCs) have received much attention for their ability to differentiate into various cell types under specific conditions and to support the proliferation of hematopoietic stem cells. However, it is unclear whether the characteristics of MSCs are altered in different disease states. In this study, we obtained and expanded MSCs from bone marrow of patients with acute lymphoblastic leukemia (ALL), Hodgkin disease (HD), and non-Hodgkin lymphoma (NHL). Our results showed that MSCs derived from ALL, HD, and NHL were similar to normal adult bone marrow-derived MSCs in morphology, growth properties, surface epitopes, and differentiation ability in vitro. Moreover, MSCs derived from ALL, NHL, and HD had a normal karyotype and ultrastructure. These cells could express hematopoietic cytokines and support hematopoiesis in long-term culture. However, adherent cells isolated from bone marrow of patients with acute myeloid leukemia (AML) showed abnormal biological properties, including heterogeneity in morphology, limited proliferation capacity, and impaired differentiation and hematopoiesis support ability. These results indicate that there are differences in the characteristics of adherent cells derived from different disease states, which may be important for reasonable MSC selection in stem cell-based therapy.     

Plasma soluble interleukin-2 receptor (sIL-2R) levels in patients with acute leukemia

The plasma soluble interleukin-2 receptor (sIL-2R) level was higher in 137 patients with acute leukemia (1,489 +/- 1,798 U/ml, including 98 cases of acute myeloid leukemia (AML), 1,063 +/- 1,414 U/ml, and 39 cases of acute lymphoblastic leukemia (ALL), 2,561 +/- 2,194 U/ml), compared to 49 normal control subjects, 421 +/- 151 U/ml). The ALL patients showed elevated plasma sIL-2R levels more frequently than the AML patients (92.3% vs 44.9%). No patient with either hypoplastic AML or AML with multilineage dysplasia and only 1 of 13 patients with acute promyelocytic leukemia (APL) had an elevated plasma sIL-2R level. All the My+ ALL patients (15 cases) showed elevated plasma sIL-2R levels. Plasma sIL-2R levels were significantly lower after chemotherapy in the ALL patients, but were not significantly lower in the AML patients. IL-2R was expressed on the leukemic cells in 36 (53.7%) of 67 AML and in 9 (21.4%) of 42 ALL cases. None of the AML M3, M4, M5, M6, or M7 subgroups showed IL-2R expression. The My+ ALL patients (42.9%, 6/14) showed IL-2R expression more frequently than the other ALL subgroups (10.7%, 3/28) (p = 0.025). The plasma sIL-2R level was correlated with the proportion of leukemic cells expressing IL-2R in acute leukemia. However, there were many cases, particularly ALL cases, who had elevated plasma sIL-2R levels without IL-2R expression on their leukemic cells. These results suggest that the plasma sIL-2R level is a valuable marker for monitoring ALL after chemotherapy, particularly in My+ ALL cases, and that the T cell immune reaction to leukemia appears to be much higher in ALL patients than in AML patients.     

Fluorescence in situ hybridization analysis of whole-arm 7;12 translocations in hematologic malignancies

Cytogenetic analysis of one case of acute myeloid leukemia (AML), one of acute lymphoblastic leukemia (ALL), one of refractory anemia with excess of blasts (RAEB), and one of acute mixed lineage leukemia (AMLL) with unbalanced 7;12 translocations mapped the breakpoints to the centromeres on both chromosomes. The rearrangements were interpreted as the whole-arm translocations der(7;12)(q10;q10) in the AML and ALL and der(7;12)(p10;q10) in the RAEB and AMLL. However, further analysis by metaphase and/or interphase fluorescence in situ hybridization (FISH) showed centric fusion only in the AML and ALL. In the RAEB and AMLL, centromeric material from chromosome 7 but not from 12 was present in the derivative chromosome. Whereas the t(7;12) resulted in loss of 12p in all four cases, the corresponding chromosome 7 imbalances differed—monosomy for 7q in the RAEB and AMLL and monosomy for 7p in the AML and ALL. Six hematologic neoplasms with unbalanced whole-arm or near-centromeric 7;12 translocations and seven dic(7;12) with juxtacentromeric breakpoints have been reported previously: 2 AML, 1 RAEB in transformation, and 10 ALL. All karyotypically informative cases had loss of 12p material. All but one of the cases with combined 7p and 12p deletion were ALL, whereas all cases with 7q and 12p loss showed myeloid differentiation. No particular clinical, morphologic, or immunophenotypic features seem to characterize ALLs with t(7;12). AMLs with an unbalanced t(7;12), often together with 5q deletions, might be associated with previous genotoxic exposure and poor prognosis.     

The activity of nucleolar organizer regions of human bone marrow cells studied with silver staining. II. Acute leukemia

The activity of nucleolar organizer regions (NOR) in chromosomes and interphase nuclei of bone marrow cells from 11 adult patients with acute lymphoblastic leukemia (ALL), 35 patients with acute nonlymphoblastic leukemia (ANLL), and eight healthy donors has been studied with silver nitrate staining. PHA-stimulated lymphocytes of the same individuals were used as standards of the maximum silver-staining patterns for each person. In 90% of patients with acute leukemia the average number of Ag+NOR in metaphases was lower when compared with that of PHA-stimulated lymphocytes. A variable expression of NOR was observed within the cell population and between individual patients. The populations tested showed high heterogeneity in relation to the content of Ag-negative mitoses. Ag+NOR per metaphase and the content of Ag-negative mitoses in bone marrow did not differ between patients with ALL and ANLL. Differences in the staining pattern in leukemic cells are discussed.     

Isolated del(14)(q21) in a case of precursor B-cell acute lymphoblastic leukemia

We present a case of del(14)(q21) as a sole abnormality in a 4-year-old boy diagnosed with precursor B-cell acute lymphoblastic leukemia (pre-B ALL). To our knowledge, this is the first case of isolated del(14)(q21) in pre-B ALL. Two pretreatment bone marrow samples obtained 5 days apart were analyzed by cytogenetics. The G-banded karyotypes of the two samples were similar, differing only in the ratio of normal/abnormal metaphases detected. Both samples showed a del(14)(q21) as the only abnormality. Fluorescence in situ hybridization performed using the probes TEL/AML1 and immunoglobulin heavy chain (IGH) showed no fusion involving the TEL and AML1 genes and only a single IGH signal in 20% of the interphase cells analyzed.     

获得性21三体恶性血液病的临床和细胞遗传学特征

目的 分析21三体恶性血液病患者的临床及细胞遗传学特点.方法 采用骨髓直接法和(或)培养法制备染色体标本,采用R显带技术进行核型分析,并进行临床随访.结果 共发现25例患者存在21三体,其中急性髓系白血病(acute myeloid leukemia,AML)13例,占同期进行染色体检查的AML患者总数的1.5%,包括M5h6例;急性淋巴细胞(acute lymphoblastic leukemia,ALL)8例,占同期进行染色体检查的ALL患者总数的2.2%,其它类型4例.25例中13例为单纯获得性21三体,其余病例均合并其它异常.随访的19例患者的中位生存期为9个月.结论 单纯21三体在AML中以M5b多见,伴21三体异常的恶性血液病预后还存在争议.     

Myelodysplastic syndrome associated with monosomy 7 in childhood: a retrospective study

Myelodysplastic syndrome (MDS) is a clonal hematopoietic stem cell disorder characterized by ineffective hematopoiesis, peripheral cytopenia, and dysplastic changes in the bone marrow. Monosomy 7 or partial loss of 7q is a common cytogenetic abnormality in MDS patients and is associated with poor prognosis. This study examined eight patients with monosomy 7 and MDS. Five MDS patients with monosomy 7 progressed to acute leukemia: three cases transformed into acute myelogenous leukemia (AML) in a mean time of only 4.6 months and two cases into acute lymphoblastic leukemia (ALL) in a mean time of 9 months. To our knowledge, this is the first report showing progression of monosomy 7 associated with MDS to ALL in the childhood period.     

XIth International Symposium for Comparative Research on Leukemia and Related Diseases

Cytogenetic and immunologic studies were performed on the cells of an 18-year-old female with ataxia telangiectasia (AT) associated with acute lymphocytic leukemia (ALL). At the onset of the leukemia 15.4% of peripheral blood cells stimulated with phytohemagglutinin (PHA) contained a tandem translocation of the long arm of chromosome #14, i.e., t(14;14). To ascertain if these karyotypically abnormal cells and the leukemic cells had a common lineage, chromosome analyses were performed on bone marrow cells. Examination of the marrow cells on the seven occasions when leukemic cells were present in the marrow, including times when they were predominant, showed only a normal karyotype without the presence of t(14;14). However, an abnormal clone, which had the karyotype 45,XX,?9,t(9;6)(q12;p13), was identified in the marrow cells on the last examination during the terminal phase of the leukemia.Immunologically, the ALL was classified as an atypical type which had characteristics in common with certain T-cell subsets. We suggest that the malignant cells did not originate from the preexisting cells with a tandem duplication of the 14q.     

Cytokine-driven differentiation of blasts from patients with acute myelogenous and lymphoblastic leukemia into dendritic cells

We investigated the ability of both acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL) blasts to differentiate into dendritic cells (DC) in vitro. Cytokine-supplemented suspension cultures of leukemic blasts in 98 patients with AML and five patients with ALL (normal karyotype, n = 2; BCR/ABL, n = 3) were performed. Mononuclear cells out of peripheral blood or bone marrow containing between 60% and 90% leukemic blasts were cultured for eight days using different growth factor combinations. The highest yield of CD1a(+)/CD14(-) cells could be obtained with stem cell factor, transforming growth factor-beta, tumor necrosis factor-alpha, GM-CSF, and FLT-3-ligand. In the AML samples the median content of CD1a(+)/CD14(-) cells after eight days of culture was 3.5% (r = 0%-82%). In five informed patients CD1a(+)/CD14(-) cells were sorted by fluorescence-activated cell sorting or immunomagnetic separation. Cytogenetic and polymerase chain reaction analyses showed known primary chromosomal aberrations (monosomy 7 and inversion 16) in the sorted fractions, respectively. Dendritic cells (DC) could be generated out of leukemic blasts in 68% of AML patients. Leukemic DC showed no phagocytosis of latex beads, but stimulated allogeneic naive cord blood-derived T cells more efficiently than did uncultured blasts. In ALL patients the median percentage of CD1a(+)/CD14(-) cells was 1.2% (r = 0.7%-3.8%) after culture. The sorted CD1(+)/CD14(-) fractions were BCR/ABL-negative when analyzed with fluorescence in situ hybridization, indicating their nonleukemic origin. Leukemic DC can be generated out of leukemic progenitors in patients with AML. These cells might become relevant for autologous and allogeneic immunotherapy in selected patients. BCR/ABL-positive lymphoblasts could not be transformed into cells with an early dendritic phenotype with the cytokines used in our experiments.     

Utility of white blood cell suspect flags and histogram pattern in the detection of acute leukemia by Advia-60 automated hematology analyzer

Blood samples from patients with acute leukemia, when analyzed with automated hematology counters, tend to introduce inaccuracies in the automated differential count and can cause diagnostic confusion without providing definite clues to the presence of abnormal cells. We designed this study to assess the utility of white blood cell (WBC) flags and histogram pattern generated by Advia-60 automated hematology analyzer in the recognition and categorization of acute leukemia. Data printouts of 31 newly diagnosed cases of acute leukemia, 22 with acute myeloid leukemia (AML) and 9 with acute lymphoblastic leukemia (ALL) were reviewed. All cases of AML and ALL generated the WBC suspect blastflag M2 associated with two of the non blast suspectflags G1 and G2. Among the cases of AML, 95.5% of the WBC histogram patterns were definitive of the presence of abnormal cells and were indicative of the myeloid nature of cells. Only 44.4% of the histograms in the cases of ALL could be definitive of the presence of abnormal cells and 33.3% were indicative of their lymphoid nature. Significantly, 55.5% of the histograms in ALL were normal. The false positives for both AML and ALL were 10.5% when only WBC flagging was considered and were reduced to 0.05% when the flags were combined with histogram patterns for interpretation. Combined flagging and histogram recognition can be of aid in identifying cases of acute leukemia and the morphologist can then assess these samples further. This ensures that cases of acute leukemia, especially in high output laboratories, are not inadvertently missed.     

Assessment of proliferative activity in leukaemic bone marrow using the monoclonal antibody Ki-67.

AIMS--To investigate proliferative activity in leukaemic and lymphomatous bone marrow infiltrates and to assess the feasibility of transport of specimens among institutions. METHODS--Proliferative activity in bone marrow trephine cryosections from 99 patients with non-Hodgkin''s lymphoma (NHL), 23 patients with acute myeloid leukaemia (AML), 11 with acute lymphoblastic leukaemia (ALL), and two with acute undifferentiated leukaemia (AUL) was investigated. Infiltration was seen in 52 out of 99 cases of NHL on bone marrow cryosections. A score was devised to assess pathological infiltrates in bone marrow trephine cryosections using the monoclonal antibody Ki-67. This method of scoring gave a measure of non-erythroid proliferative activity. RESULTS--Mean Ki-67 positivity in bone marrow infiltrates in 31 low grade B cell lymphomas (Kiel classification) was 0.3% before and 4.7% after treatment, 16.4% in seven high grade B cell lymphomas, and 17.8% in 12 peripheral T cell lymphomas. In 48 cases of NHL, bone marrow cryosections had not been infiltrated, and in all but one case the percentage of Ki-67 positive cells in normal marrow was less than 3%; the remaining case showed coexistent myelodysplasia and 8% bone marrow Ki-67 positivity. In eight cases of common ALL at diagnosis, the mean Ki-67 positivity in marrow cryosections was 24.9%, significantly higher than the 2.4% Ki-67 positivity seen in AML (p < 0.05). One of the two cases of common ALL with less than 1% Ki-67 positivity was refractory to treatment. CONCLUSIONS--Proliferative activity of erythroid elements in the bone marrow varies greatly. Immunostaining of bone marrow cryosections using Ki-67 permits accurate assessment of non-erythroid proliferative activity in lymphomas and leukaemia. High grade B cell lymphomas and peripheral T cell lymphomas invading the marrow have very similar mean proliferative activities. Such levels of proliferation are of the same order as those seen in common ALL, but much higher than those seen in AML.     

Nonrandom cytogenetic changes in New Zealand patients with acute myeloid leukemia

Bone marrow clones with abnormal chromosomes were observed in 56% of 66 patients with forms of acute myeloid leukemia [French-American-British (FAB) M1-M6]. Acute myeloblastic leukemia (AML, M1 and M2) was the most common form, and 65% of these patients showed chromosomal abnormalities compared with 41% of patients with acute myelomonocytic leukemia (AMMoL, M4). The recognized nonrandom chromosomal abnormalities found were trisomy 8, monosomy 5 or 7, trisomy 1q, t(6;9), t(8;21), t(15;17), and abnormalities in 17q. There was also a strong involvement of chromosome No. 11: Abnormalities were found in eight patients when their leukemia was diagnosed and in a further three patients during the course of karyotypic evolution. Six of these patients had AMMoL or AMoL. Complex or multiple clones were found in 37% of AML patients at diagnosis. Our AML patients had a reduced frequency of abnormalities in chromosome No. 5 or 7 and an increased frequency of abnormalities in chromosome No. 8 compared with studies reported in other countries (p = 0.01). This difference suggests that in New Zealand AML might be caused by factors different from those operating in more industrialized centers.     

The use of fluorodeoxyuridine synchronization for cytogenetic investigation of acute lymphoblastic leukemia.

The role of fluorodeoxyuridine (FUdR) synchronization in cytogenetic analysis of acute lymphoblastic leukemia (ALL) was investigated using samples of bone marrow (BM) (10 patients) and peripheral blood (PB) (2 patients), prepared for chromosome analysis using both 24-hour unstimulated cultures (24-hr) and cultures synchronized with FUdR. The mitotic index (MI) in FUdR was lower than in 24-hr in 8 of 10 BM and 2 of 2 PB cultures. The quality of the metaphases was the same in both cultures. The FUdR had a lower percentage of abnormal cells than the 24-hr in the 7 BM samples with a normal/abnormal population and sufficient analyzable cells in each culture for comparison (p less than 0.05). PB FUdR cultures yielded only normal cells. We conclude that FUdR cultures are inferior to 24-hr cultures for chromosome analysis in ALL.