STUDY ON AUTOINHIBITORY ACTIVITY OF LYMPHOID LEUKEMIA

Autoinhibitory activity has been discovered in murine T lymphocyte leukemia models derived from 615 mice in our lab. It was designated 615 mice leukemia associated inhibitor (LAI-615) . To further confirm whether LAI activity could be found in human leukemia, 6 ALL cases and 2 AML cases were examined. The results showed that 5/6 of ALL and 1/2 of AML cases had detectable LAI activity. The different sensitivities of LAI activity were also found between autologous bone marrow cells and human leukemic cell lines, which indicate that autoinhibitory activity might have individual specificity.     

用骨髓细胞培养净化法对白血病人进行自体骨髓移植

对白血病人的骨髓细胞进行长期培养可以选择性地对白血病细胞抑制,而有利于正常造血组细胞的增长。我们利用此特点进行对白血病人自身骨髓移植前的骨髓净化。对5例白血病人进行了ABMT,ALL2例,AML1例,CML(CP)1例,CML(AP)1例。预处理     

t（8;21）（q22;q22）急性髓系白血病转化为急性淋巴细胞白血病一例并文献复习

目的 探讨t（8; 21） （q22; q22）急性髓系白血病（AML）转化为急性淋巴细胞白血病（ALL）的克隆转化机制.方法 报道1例初诊为t（8; 21）AML,后转化为ALL患者的临床资料.期间对其进行持续细胞形态学、遗传学和分子生物学监测.结果 患者人院时诊断为t（8;21）AML,AML1-ETO融合基因阳性,治疗后达到完全缓解.半年后转化为ALL,诱导化疗后再次缓解.结论 t（8;21）（q22;q22）AML转化的ALL可能起源于具有髓/淋分化潜能的多能造血干细胞,早期化疗抑制了占优势地位的髓系白血病克隆,从而使具有不同表型的淋系亚克隆增殖.     

恶性血液病患者周围血细胞中腺苷脱氨酶活性

作者测定了正常人和恶性血液病患者的淋巴细胞、粒细胞和红细胞中腺苷脱氨酶(ADA)活性,并探讨了 ADA 与恶性血液病的关系。材料:正常人组22例,年龄23～60岁。恶性血液病组45例,包括慢性淋巴细胞性白血病(CLL)9例、白血病性网状内皮细胞增生症2例,急性淋巴肉瘤细胞性白血病1例、急性淋巴细胞性白血病(ALL)7例、急性粒细胞性白血病(AML)6例、何杰金氏病7例、非何杰金氏病淋巴瘤10例和多发性骨髓瘤3例。ALL 中五例为儿童,余皆成人。     

急性淋巴细胞白血病AML1基因重排与MLL基因丢失同时出现临床意义的探讨

目的:对急性淋巴细胞白血病(ALL)急性髓系白血病(AML1)基因重排与混合谱系白血病(MLL)基因丢失同时出现进行探讨。方法:在常规细胞遗传学(CC)分析基础上运用荧光原位杂交技术(FISH),采用多种位点特异性DNA探针(染色体全染、特殊位点和双色易位融合探针),对63例ALL患者(8例成人,55例儿童)进行分析。结果:63例ALL患者中有4例(6.3%)出现MLL基因重排,其中3例出现MLL基因丢失,1例出现MLL基因的移位即t(4;11),3例出现了MLL基因丢失的患者同时合并有AML1基因重排,2例为t(12;21)易位而形成的TEL/AML1融合基因,1例为AML1基因复制引起的环形21号染色体,即r(21)。55例儿童ALL中有15例(27.3%)出现AML1基因重排,即由t(12;21)易位而形成的TEL/AML1融合基因,TEL/AML1阳性患者免疫分型均为B细胞型,8例成人均无t(12;21)。结论:儿童ALL常合并有t(12;21),TEL/AML1融合基因的出现是预后良好的指标,而MLL基因重排的患者具有对常规化疗不敏感及预后不良的特点,两者同时出现说明白血病染色体重排、病理过程及影响预后因素的复杂性。     

抗原处理相关转运因子在小儿急性白血病患者中的表达及其临床意义

背景与目的:抗原处理相关转运因子(transporterassociatedwithantigenprocessing,TAP)参与免疫监视,因而可能与肿瘤发生有关。本文旨在探讨急性白血病TAP分子表达及其临床意义,探讨急性白血病的治疗策略。方法:采用RT-PCR检测34例初治急性淋巴细胞白血病(acutelymphoblasticleukemia,ALL)(初治组)、15例复发ALL(复发组)及20例急性髓系白血病(acutemyeloidleukemia,AML)患者骨髓中TAP亚单位TAP1和TAP2的表达。20例无全身性疾病外科住院患儿作为对照组。使用数码成像分析仪测定并计算扩增条带的相对于阳性内对照GAPDH的吸光度(A)值。结果:ALL初治组和ALL复发组的TAP1A值(分别为0.448±0.167和0.169±0.021)及TAP2的A值(分别为0.196±0.180和0.112±0.020)均低于对照组,P均<0.01;AML组TAP2的A值低于对照组(P<0.01);ALL复发组TAP1的A值低于ALL初治组,P<0.05;ALL初治组复发者(6/34)TAP1的A值(0.215±0.159)较持续完全缓解(constantcompleteremission,CCR)者低(24/34,0.462±0.189,P<0.05)。结论:小儿ALL和AML均存在TAP分子低表达,这可能促使白血病细胞逃避免疫监视;TAP1亚单位低表达可能与ALL的复发有关。     

自体骨髓移植治疗急性白血病15例临床分析

我院自1988年11月以来,利用微波净化或未净化的自体骨髓移植(ABMT)治疗急性白血病15例,取得了良好疗效。微波净化组10例,其中急性淋巴细胞白血病(ALL)6例,急性髓性白血病(AML)4例。未净化组5例,其中ALL2例,AML3例。所有病人皆住层流无菌病室隔离护理和适当支持治疗。采集骨髓4℃保存50～56h,然后回输给病人。预处理方案为全     

米托蒽醌联合治疗儿童难治性复发性急性白血病的临床疗效观察

 目的 观察总结米托蒽醌（MTZ）联合治疗儿童难治性复发性急性白血病（RRAL）的疗效。方法 RRAL 12例,急性淋巴细胞白血病（ALL）9例,急性髓性白血病（AML）3例;ALL用VMLP／Dex方案（长春新碱、米托蒽醌、左旋门冬酰胺酶、泼尼松／地塞米松）2～4周;AML用MAE方案（米托蒽醌、阿糖胞苷、依托泊苷）或大剂量阿糖胞苷+依托泊苷。结果 9例ALL,CR 7例,PR 1例,1例未复查;3例AML,CR 2例,NR 1例。总CR率为81.8 %（9／11）,总有效率90.9 %（10／11）。用药后骨髓抑制较明显,大部分病例ANE≤0.1×109／L持续1～2周,轻微肝功能损害,未见药物相关的心脏损害。结论 MTZ不失为治疗儿童难治性复发性急性白血病的有效药物之一,用药后要注意预防和治疗骨髓抑制后出现的各种感染和出血。     

序贯性大剂量阿糖胞苷合并氨甲喋呤治疗难治性急性白血病

作者运用大剂量阿糖胞苷(AraC)合并氨甲喋呤(MTX)序贯性给药(S-HAM)方法治疗22例难治性急性髓细胞白血病(AML)及急性淋巴细胞白血病(ALL),评价其临床疗效及副作用。 22例难治性白血病病人,男11例,女11例,其中AMC 18例,ALL 4例,年龄17～66岁,平均37岁。AML、ALL按FAB协作组诊断标准诊断。难治性急性白血病的诊断标准为:①AML或ALL经常规方案化疗两个疗程无效;②第一次完全缓解后6个月以内复发;③第一次完全缓解后超过6个月以后复发,但对原诱导方案无效;④复发两次或两次以上。     

11q23染色体异常的恶性血液病的细胞遗传学和临床特征

目的研究11q23异常与恶性血液病的形态学、细胞遗传学及预后的关系。方法采用骨髓直接法和(或)培养法制备染色体标本,用R显带技术,对887例急性髓系白血病(AML)、370例急性淋巴细胞白血病(ALL)及其它类型恶性血液病进行核型分析。结果共发现21例具有11q23异常的恶性血液病患者,其中AML 10例,ALL 9例,在AML和ALL中的发病率分别为1.1%和2.4%。10例AML中,6例为急性单核细胞白血病。异常核型有7种:del(11)8例;t(4;11)6例;t(1;11)3例;t(2;11)1例;t(5;11)1例;t(11;14)1例;t(11;19)1例。6例t(4;11)均为急性淋巴细胞白血病。随访的14例患者中位生存期为274天。结论11q23异常主要见于急性单核细胞白血病和急性淋巴细胞白血病患者,其临床预后不良。     

TEL-AML1, expressed from t(12;21) in human acute lymphocytic leukemia,induces acute leukemia in mice

TEL-AML1 is expressed from the t(12;21) chromosomal translocation inB-precursor acute lymphocytic leukemia (ALL). Creation of the TEL-AML1fusion disrupts one copy of the TEL and AML1 genes, and loss of TEL or AML1 is also associated with cases of acute leukemia without TEL-AML1. To determine whether TEL-AML1 can contribute to leukemogenesis, we transduced marrow from C57BL/6 mice with a retroviral vector expressing TEL-AML1 or with a control vector. Transduced cells were introduced into irradiated syngeneic recipients. Two of 9 TEL-AML1 mice developed ALL (one T-lineage ALL and one B-precursor ALL), whereas 0 of 20 control mice developed leukemia. The B-precursor ALL was retransplantable and expressed TEL-AML1. We similarly transduced marrow from C57BL/6 mice lacking the overlapping p16(INK4a)p19(ARF) genes and transplanted the cells into wild-type recipients. No control mice died, but six of eight TEL-AML1/p16p19 mice died with leukemia. Overall, these findings indicate that TEL-AML1 contributes to leukemogenesis and may cooperate with loss of p16(INK4a)p14(ARF) to transform lymphoid progenitors.     

Amplification and rearrangement of proto-oncogene c-abl in human leukemia cells

DNA of peripheral blood or bone marrow leukocytes from 8 normal subjects, 7 cases of acute lymphocytic leukemia (ALL), 2 of acute myelogenous leukemia (AML) and 1 of chronic myelogenous leukemia (CML), having been digested by endonuclease Eco RI or Pst I separately, was hybridized with the probes of 3' fragment (Pst I/Hind III) or 5' fragment (Hinc II/Pst I) of Abelson murine leukemia virus (A-MuLV) oncogene v-abl. The proto-oncogene c-abl, which is homologous to v-abl, was found amplified in 4 ALL, 1 CML and 1 AML. In one of these 4 ALL, c-abl was amplified even over 100 times. A new c-abl BamH I fragment with 6.7 kilobase pairs (kb) in length was observed in 2 ALL and 1 CML out of these 6 cases with amplification, but none of this fragment was found in the normal subjects or other leukemia patients. These 3 patients with the presence of 6.7 kb fragment were high risk ones and 2 of them had died, suggesting that 6.7 kb fragment be the index of poor prognosis. The amplification and rearrangement of c-abl imply the activation of proto-oncogene in leukemogenesis.     

Molecular Characterization of FLT3 Mutations in Acute Leukemia Patients

Fms-like tyrosine kinase 3 (FLT3) performs a vital role in the pathogenesis of hematopoietic malignancies.Therefore in recent times, the focus of several studies was on use of FLT3 as a prognostic marker. The presentstudy investigated the molecular characterization and incidence of FLT3 mutations in acute leukemia patients inPakistan. A total of 55 patients were studied, of which 25 were suffering from acute lymphoblastic leukemia (ALL)and 30 were suffering from acute myeloid leukemia (AML). The polymerase chain reaction demonstrated FLT3/ITD mutations in 1 (4%) of 25 ALL patients, a male with the L2 subtype. In AML cases the rate was 4 (13.3%) of30, three males and one female. The AML-M4 subtype was found in three and the AML M2 subtype in the other.In the AML cases, a statistically significant (p=0.009) relationship was found between WBC (109/L) and FLT3/ITD positivity. However, no significant relationship was found with other clinical parameters (p>0.05). In acutemyeloid leukemia (AML) FLT3/ITD+ mutation was more prevalent in elderly patients 31-40 age groups, 21-30and 51-60 age groups respectively. In acute lymphoblastic leukemia (ALL) statistically no significant relationshipwas found between clinical features and FLT3/ITD positivity (p>0.05). However, in acute lymphoblastic leukemia(ALL) FLT3/ITD+ mutation was more commonly found in age groups of 21-30.     

Detection of acute leukemia cells with mixed lineage leukemia (MLL) gene rearrangements by flow cytometry using monoclonal antibody 7.1.

Translocations involving 11q23 are among the most common genetic abnormalities in hematologic malignancies, occurring in approximately 5-10% of acute lymphoblastic leukemia (ALL) and 5% of acute myeloblastic leukemia (AML). In 11q23 translocations, the mixed lineage leukemia (MLL) gene on chromosome 11, band q23, is usually disrupted. The human homologue of the rat NG2 chondroitin sulfate proteoglycan molecule, as detected by the monoclonal antibody (moab) 7.1, was shown to be expressed on leukemic cells with MLL rearrangements of children with acute leukemia. We further investigated the reactivity of the moab 7.1 on 533 cell samples of adults (n = 215) and children (n = 318) with acute leukemias (271 AML, 217 B-lineage ALL, 37 T-lineage ALL, eight CD7+ CD56+ myeloid/natural killer cell precursor acute leukemias) by flow cytometry. In AML, 38 samples were positive for moab 7.1 ('20%-cut-off-level'). These moab 7.1-positive AML cases revealed a myelomonocytic-differentiated immunophenotype with coexpression of the NK cell marker CD56 in 33 of 38 cases. Two of eight cell samples of the recently described CD7+ CD56+ myeloid/natural killer cell precursor acute leukemia entity reacted with moab 7.1. In ALL, 35 samples mostly of the pro-B-ALL subtype (33 pro-B-ALL, one common-ALL, one pre-B-ALL) were positive for moab 7.1. 58 (81%) of 72 samples with MLL rearrangements were positive for moab 7.1 including 28/31 with a t(4;11), 16/17 with a t(9;11), 3/5 with a t(11;19), and 2/6 with a del(11)(q23). All moab 7.1-positive ALL (n = 34) and childhood AML (n = 17) cases revealed MLL rearrangements as detected by Southern blot analysis and RT-PCR. However, 11 adults with AML, and one adult with moab 7.1-positive CD7+ CD56+ myeloid/natural killer cell precursor acute leukemia were negative for MLL rearrangements as proved by Southern blot analysis. We conclude that moab 7.1 is a sensitive but not entirely specific marker for the identification of 11q23-associated AML and ALL by flow cytometry in children and adults.     

多元耐药基因（MDRI）在急性白血病细胞中表达的意义

观察多元耐药ＭＤＲ１基因在急性白血病细胞中的表达及其意义。方法取１５例急性白血病（ＡＭＬ１２例，ＡＬＬ３例）患者的骨髓细胞通过ＲＴＰＣＲ等法，检测ＭＤＲ１，分析其表达情况以及与临床治疗效果。结果９／１５例（ＡＭＬ８例，ＡＬＬ１例）的ＭＤＲ１表达阳性（阳性率６０％）。其中６／９例经化疗不缓解。１／９例部分缓解，２／９例完全缓解，在ＭＤＲ１表达阴性者中，５／６例完全缓解，１／６例不缓解。结论急性白血病患者的ＭＤＲ１基因表达和化疗的效果有密切联系。     

Soluble Interleukin-2 Receptor, Soluble CD8 and Soluble Intercellular Adhesion Molecule-1 Levels in Hematologic Malignancies

Plasma levels of soluble interleukin-2 receptor (sIL-2R), soluble CD8 (sCD8) and soluble intercellular adhesion molecule 1 (sICAM-1) were determined by ELISA assays in about 100 patients with hairy cell leukemia (HCL), acute myelomonocytic leukemia (AMMoL), acute myelocytic leukemia (AML), chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), acute lymphoblastic leukemia (ALL), adult T-cell leukemia (ATL), and mycosis fun-goides (MF). Additionally, cultured AML, ALL, and CLL cells grown with and without 12-0-tetra-decanoyl-phorbol-13-acetate (TPA) were tested for IL-2R (CD25) expression by indirect immunofluorescence. Supernatants of these cultures were also tested for sIL-2R by ELISA.

Elevated sIL-2R levels were found in HCL patients at initial diagnosis and relapse, in AMMoL, in AML, in the accelerated and non-accelerated phases of B-CLL, in PLL, in non-T/non-B ALL, in B-ALL, in mixed lineage ALL, in T-CLL, in T-ALL, and in active MF. Reduced levels of sIL-2R were encountered in HCL patients in remission, in pre-T-ALL, and in MF patients in remission. Also, in non-accelerated CLL sIL-2R levels were less elevated than in later stages of the disease. In T-CLL, sIL-2R was only slightly elevated. Thus, we believe sIL-2R could prove to be a useful marker of disease stage, subtype, and prognosis in several hematologic malignancies.

The cultures with and without TPA suggested that the undetermined source of sIL-2R in HCL, ALL and AML could indeed be the malignant cells but perhaps not so in the case of B-CLL.

Plasma sCD8 was found to be below normal control levels in HCL, and lowest in relapsing cases. In addition, sCD8 levels were below normal in pre-T-ALL, and in MF. Levels in the non-accelerated phase of B-CLL approximated those of controls. Elevated levels of sCD8 were observed in AML, AMMoL, accelerated stage B-CLL, PLL, non-T/non-B ALL, B-ALL, mixed lineage ALL, T-ALL, T-CLL, and ATL. Thus, in a few instances, sCD8 may also correlate with disease subtype, as well as stage. Although sICAM-1 levels were elevated in all leukemias, its levels in CLL did not appear to be related to disease activity. Whether this is true or not for other leukemias would require additional work on sICAM-1 levels and its relationship to disease activity and prognosis.     

Proteomic analysis of human acute leukemia cells: insight into their classification.

PURPOSE: French-American-British (FAB) classification of acute leukemia with genetic heterogeneity is important for treatment and prognosis. However, the distinct protein profiles that contribute to the subtypes and facilitate molecular definition of acute leukemia classification are still unclear. EXPERIMENTAL DESIGN: The proteins of leukemic cells from 61 cases of acute leukemia characterized by FAB classification were separated by two-dimensional electrophoresis, and the differentially expressed protein spots were identified by both matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and tandem electrospray ionization MS (ESI-MS/MS). RESULTS: The distinct protein profiles of acute leukemia FAB types or subtypes were successfully explored, including acute myeloid leukemia (AML), its subtypes (M2, M3, and M5) and acute lymphoid leukemia (ALL), which were homogeneous within substantial samples of the respective subgroups but clearly differed from all other subgroups. We found a group of proteins that were highly expressed in M2 and M3, rather than other subtypes. Among them, myeloid-related proteins 8 and 14 were first reported to mark AML differentiation and to differentiate AML from ALL. Heat shock 27 kDa protein 1 and other proteins that are highly expressed in ALL may play important roles in clinically distinguishing AML from ALL. Another set of proteins up-regulated was restricted to granulocytic lineage leukemia. High-level expression of NM23-H1 was found in all but the M3a subtype, with favorable prognosis. CONCLUSIONS: These data have implications in delineating the pathways of aberrant gene expression underlying the pathogenesis of acute leukemia and could facilitate molecular definition of FAB classification. The extension of the present analysis to currently less well-defined acute leukemias will identify additional subgroups.     

Amplification of AML1 gene is present in childhood acute lymphoblastic leukemia but not in adult, and is not associated with AML1 gene mutation.

The AML1/CBFA2/RUNX1 gene is the target of many recurrent translocations seen in different leukemia subtypes. The t(12;21)(p13;q22) is the most frequent translocation observed in childhood B acute lymphoblastic leukemia (ALL), occurring in 20% to 25% of cases. In adult ALL this rearrangement is scarce. Another route of AML1deregulation could be point mutations in the runt domain. We now report on AML1amplification in two cases of childhood ALL, found in a series of 107 consecutive children with B-lineage ALL analyzed by fluorescence in situ hybridization (FISH). A parallel analysis of 42 adult B-ALL failed to detect any AML1 rearrangement by FISH. The two patients with AML1 amplification were further analyzed using molecular techniques. SSCP analysis did not detect any mutation. Furthermore, direct sequencing of the cDNA did not reveal any mutation. In conclusion, AML1amplification seems to be observed only in childhood ALL and is not associated with AML1 gene mutation. Other mechanisms, such as gene dosage effects could be hypothesized.     

CD45设门多参数流式细胞术在急性白血病免疫表型分析中的应用

目的：分析急性白血病的抗原表达及其临床意义。方法：采用一组系列相关单抗直接免疫荧光标记CD45设门的多参数流式细胞术,检测35例急性白血病患者的免疫表型。结果：11例ALL中B－ALL8例,T－ALL3例,其中出现髓系抗原表达4例,占36．4％,CD34表达10例,占90．1％;24例AML中伴淋系抗原表达7例,占29．17％,CD34表达16例,占70．8％,DR的表达与CD34一致,5例M3患者均无CD34和HLA DR表达。伴髓系统原表达的ALL CR率低于髓系抗原阴性表达者（1／3及5／6）,但统计学上差异无显著性（P＞0．05）;伴淋系抗原表达的AML患者CR率明显低于淋系抗原阴性表达者（0／5及10／10）,两组间差异具有显著性（P＜0．01）。结论：CD45设门的多参数流式细胞术是分析白血病免疫表型的最好方法,白血病抗原的错义表达是预后不良的因素之一。