急性髓系白血病M2型免疫表型及细胞遗传学研究

目的 探讨急性髓系白血病M2型的免疫表型及细胞遗传学特征.方法 采用一组单克隆抗体和三色流式细胞术对112例M3患者进行免疫表型及核型分析.结果 112例M3患者中,70例(62.5%)表达干细胞标志CD34.髓系标志中以CD13(87.5%)M2患者表达最高,其次是CD33(83.9%)和CD14(2.7%).分别有34例(30.4%)及24例(21.4%)患者表达T系及B系标志.112例M2患者中,73例(65.2%)检出异常核型,共检出28种数目异常,25种结构异常.47例(42.0%)患者检出t(8;21).CD19及CD34单独或共同表达在核型异常及伴t(8;21)的M2患者中显著增高(P＜0.05).结论 M3以髓系抗原表达为主,部分有淋系抗原表达,CD19及CD34表达与t(8;21)显著相关.     

急性髓系白血病M2型免疫表型及细胞遗传学研究

 目的　探讨急性髓系白血病M2型的免疫表型及细胞遗传学特征。方法　采用一组单克隆抗体和三色流式细胞术对112例M2患者进行免疫表型及核型分析。结果　112例M2患者中,70例（62.5 %）表达干细胞标志CD34。髓系标志中以CD13（87.5 %）M2患者表达最高,其次是CD33（83.9 %）和CD14（2.7 %）。分别有34例（30.4 %）及24例（21.4 %）患者表达T系及B系标志。112例M2患者中,73例（65.2 %）检出异常核型,共检出28种数目异常,25种结构异常。47例（42.0 %）患者检出t（8;21）。CD19及CD34单独或共同表达在核型异常及伴t（8;21）的M2患者中显著增高（P＜0.05）。结论　M2以髓系抗原表达为主,部分有淋系抗原表达,CD19及CD34表达与t（8;21）显著相关。     

成人急性髓细胞性白血病免疫表型与细胞遗传学及临床特征的关系

背景与目的:新的WHO分类已迅速应用于白血病的诊断。依据多个系相关抗原的表达,多参数高分辨流式细胞术可准确地识别白血病细胞的系列来源和分化阶段,而且某些抗原表达与细胞遗传学改变和预后密切相关。本研究旨在探讨初治成人急性髓细胞性白血病(acutemyeloidleukemia,AML)的免疫表型特征,并对其与FAB分类、细胞遗传学和临床表现的关系进行分析。方法:采用多参数高分辨流式细胞术对96例成人AML患者骨髓进行免疫表型分析,染色体G显带技术对其中的73例进行核型分析。结果:AML患者中,某些免疫表型特征与FAB分类具有相关性,包括M3中缺乏表达HLA鄄DR、CD34和CD56,但CD2的表达增加;M2中CD19、M5中CD14和CD56的表达增加,而M0中未见MPO的表达。本组AML核型异常率为54.8%,其中CD22、CD56和TdT的表达与核型异常有显著性相关。10例t(8;21)改变仅见于M2中,并高表达CD15、CD19、CD34和CD56,但未见CD2和CD7表达。7例伴t(15;17)的M3患者中未见淋系抗原的表达。此外,CD4和TdT抗原的表达与患者年龄、CD7和CD14的表达与外周血白细胞计数、CD4、CD14和CD56的表达与血小板计数等均有显著性正相关。结论:AML患者免疫表型与细胞遗传学的相关性提示AML抗原的异常表达可能与基因的异常改变密切相关。白血病免疫表型的检测有助于     

CD_4抗原在急性髓系白血病细胞上的表达及其意义

目的 :探讨 CD4抗原在急性髓系白血病细胞上的表达及意义。方法 :对 82例急性髓系白血病患者进行免疫表型、细胞遗传学分析。结果 :CD4在 AML患者中表达率为 40 .2 % ,M5 中阳性率最高 92 .9% ,M4次之为5 5 % ,CD+4 AML 高表达 HL A- DR,CD3 8、CD3 3 、CD1 5 、CD1 4 、TB系列抗原阴性。CD+4 AML 中可见 11q2 3、inv(16 )、t(9;2 2 ) ,未见特异性染色体异常。但伴 11q2 3和 inv(16 )异常的 AML频繁表达 CD4,阳性率分别为 86 .3%、6 0 .2 %。CD+4和 CD- 4 AML 在年龄、性别、肝脾肿大、CNS- L、DIC、1疗程 CR率等临床特征方面无明显不同。结论 :CD+4 AML 是一组起源较早的具有单核细胞特征的髓系白血病 ,CD4的表达对 AML- M4、M5 尤其是 M5 亚型的鉴别诊断有重要价值     

急性粒-单核细胞白血病的细胞遗传学异常研究

背景与目的：2001年WHO分型特别提出4种伴再现性遗传学异常的急性髓细胞白血病（AML），其中inv（16）（p13：q22）与急性粒一单核细胞白血病（M4）密切相关，是预后好的标志。本研究旨在探讨M4的细胞遗传学特征。方法：采用直接法及短期培养法制备骨髓细胞染色体，并以R显带技术对89例M4患者进行核型分析，并应用间期荧光原位杂交（I-FISH）技术对其中伴有＋22异常的患者进行inv（16）检测。结果．89例M4患者中，异常染色体检出率为40．4％（36／89），共12种主要异常核型，其中5种为特异性染色体异常，见于25例患者，占核型异常患者的69．4％。单纯＋8（10例）为最常见的数目异常；结构异常最多见的是inv（16）（5例）；t（8；21）者3例；伴t（9；22）者有1例；其中5例inv（16）及3例＋22均只见于M4Eo患者。3例＋22患者FISH检测inv（16）均阳性。结论：细胞遗传学研究对于急性粒一单核细胞白血病的诊断具有重要价值，但是细胞遗传学对inv（16）检测阳性低，对怀疑病例及所有＋22异常的患者，应尽可能进行FISH检测。     

t(8;21)急性髓系白血病患者MICM分型及预后的回顾性分析

 目的　分析t(8;21)急性髓系白血病（AML）患者的细胞形态学、免疫表型、遗传学、分子生物学（MICM）分型及临床治疗疗效。方法　运用瑞特染色法、FAB细胞形态分类标准、流式细胞术（FCM）直接免疫荧光标记技术、遗传学染色体吉姆萨显带技术及RT-PCR技术对70例确认有t(8;21)与AML1-ETO融合基因双阳性的AML患者及70例正常染色体核型的AML患者进行分析和比较。结果　70例t(8;21)AML患者中M1 1例,M2 64例,M4 3例,无法分型的急性白血病（AL）2例;免疫表型分析发现CD13、CD33、CD34、CD117高表达,40 %表达CD19,11 %表达CD15,10 %表达CD11b,7 %表达CD7;遗传学显示50 %的t(8;21) AML患者有附加染色体异常,主要为性染色体丢失、9q-及超二倍体;RT-PCR检测AML1-ETO融合基因100 %阳性。CD+19 t(8;21) AML患者完全缓解（CR）率72 %,CD+19伴CD+7 t(8;21)AML患者CR率为0,正常核型CR率31 %。结论　t(8;21) AML患者主要在M2中集中出现,附加染色体异常较多见。CD19表达较高,而CD7表达极低,CD34、CD117高表达,这些抗原的表达可能与核型密切相关。CD+19 是预后良好的指标,但同时出现CD+7,则预后不良。     

232例急性髓细胞白血病免疫表型及与细胞遗传学关系分析

目的探讨初治成人急性髓细胞性白血病（acute myeloid leukemia,AML）的免疫表型特征,并对其与FAB分类、细胞遗传学的关系进行分析。方法采用CD45/SSC双参数散点图设门方法、三色流式细胞术对232例急性髓细胞白血病细胞进行免疫表型分析,染色体G显带技术对其中的172例进行核型分析。结果AML患者中,CD38、CD33和CD13的表达最常见,CD117、CD11b分别有助于区分髓系和淋系白血病,在淋系抗原中,以CD9、CD7较常见。CD7可能是一个独立的预后因素。某些免疫表型特征与FAB分类具有相关性,包括M3中缺乏表达HLA-DR和CD34,而CD9表达增加。CD11b有助于M5与M1、M2、M3的区分,M5中CD14的表达率增加,其阳性率明显高于M2。CD15可能有助于M5亚型的鉴别。本组AML核型异常率为71.6%,t（8;21）（q22;q22）主要见于M2（79.0%）,还见于M5。单纯t（8;21）（q22;q22）易位10例（52.6%）,伴其它染色体异常的为9例（47.4%）,主要附加异常染色体类型为性染色体的丢失。CD13、CD9的表达与染色体异常显著性相关。CD117、CD15同时表达的AML少有预后较差染色体的异常。伴t（8;21）（q22;q22）异常的AML表达CD15、CD33、CD38显著增加。结论白血病免疫表型的检测有助于AML的诊断和分类。免疫表型与细胞遗传学的相关性提示AML抗原的异常表达可能与基因的异常改变密切相关。     

成人急性淋巴细胞白血病免疫表型及细胞遗传学特征分析

目的：对110例成人急性淋巴细胞白血病（ALL）患者进行免疫表型及细胞遗传学特征分析.方法：采用CD45/SSC参数设门四色流式细胞术检测110例成人ALL患者治疗前骨髓细胞的免疫表型,染色体R显带技术对其中的73 例进行核型分析.结果：110例ALL患者中,21.8%为T-ALL,78.2%为B-ALL.47.3%的ALL患者表达髓系抗原 （MyAg）,CD13是成人ALL中最常见的MyAg （32.1%）.T-ALL髓系相关抗原表达总的阳性率（45.8%）与B-ALL（47.7%）无统计学差异.可供核型分析的73例中核型异常者37例（50.7%）,最常见的遗传学异常为Ph染色体,占23.3%.结论：免疫表型对ALL的诊断与分型至关重要,免疫表型与患者的异常核型改变及临床特征关系密切.     

双色间期荧光原位杂交检测急性粒-单核细胞白血病inv(16)

 目的 探讨双色荧光原位杂交（D-FISH）技术在检测急性粒-单核细胞白血病（M4）inv（16）中的价值。方法 用荧光素直接标记的双色断裂点分离的基因探针CBFβ对42例M4患者进行间期D-FISH检测,并与常规细胞遗传学进行比较分析。结果 采用R显带核型分析的42例M4患者均未发现inv(16),而D-FISH检测发现10例（23.8 %）inv(16),阳性率为30.1 % ~ 98.0 %,平均阳性率82.1 %。5例M4Eo患者均伴有inv(16)。10例阳性患者中7例核型正常,2例伴+22,1例伴+8。结论 inv(16)在M4中阳性率为23.8 %,与M4Eo密切相关;用CBFβ双色探针进行D-FISH为检测inv(16)提供了有效的方法,是细胞遗传学检查的重要补充。     

急性白血病首次复发后免疫表型及细胞遗传学改变

目的 :分析成人急性白血病首次复发前后免疫表型和细胞遗传学改变。方法 :采用间接荧光法分析急性白血病患者的免疫表型 ,用 G分带技术研究患者的染色体核型。结果 :5例中 4例急性淋巴细胞白血病 ( ALL )复发后的患者发生免疫表型变化 ,均涉及 CD3 4 抗原表达 ;CD3 4 抗原的表达与再诱导治疗结果呈负相关。 10 /12例急性髓细胞白血病 ( AML )复发患者免疫表型发生改变 ,涉及 CD1 4 、CD1 5 、CD1 3 、CD3 3 、CD3 4 、CD7、HLA-DR等抗原 ;CD1 4 、CD3 3 、CD3 4 发生改变的患者再诱导治疗效果差 ;CD1 5 由阴性转为阳性的患者再诱导效果较好。 7例患者复发前后均进行了染色体核型分析 ,4例出现主要的染色体核型改变 ,其中 3例为异常核型转为正常核型 ;染色体核型发生改变的病例再诱导治疗效果均较差。结论 :成人急性白血病复发后免疫表型和细胞遗传学均可发生改变 ,这些改变与再诱导治疗的结果有关。     

Clonal chromosomal abnormalities in the stem cell compartment of patients with acute myeloid leukemia in morphological complete remission.

Acute myeloid leukemia arises from the clonal expansion of a malignant transformed progenitor cell. Despite intensive chemotherapy, final disease eradication is achieved by a small proportion of cases only and 50-70% of adults with AML will ultimately relapse and die from their disease. Hence residual disease below the level of morphological detectability must be assumed in clinical and morphological complete remission. CD34+/CD38- and CD34+/CD38+ subpopulations of seven patients in morphological complete remission were isolated by FACS (purity >98%) and were analyzed by conventional cytogenetics or FISH for chromosomal aberrations. In five of seven patients, clonal chromosomal abnormalities were detected in the CD34+/CD38+ subpopulation and in one patient with AML M2 (add (2)(q37)) in the most immature CD34+/CD38- stem cell compartment. One patient with AML M4Eo (inv(16),+8), showed a normal karyotype by conventional cytogenetic analysis, whereas four of 15 metaphases of the sorted CD34+/CD38+ subpopulation revealed the inversion 16. These observations underline that leukemic cells can survive intensive chemotherapy in the niche of the stem cell compartment. In some patients the sensitivity for the detection of persistent leukemic cells seems to be higher in FACS-sorted subpopulations than conventional cytogenetic analysis of the unseparated bone marrow. Immunophenotyping revealed minimal residual disease in four of the patients. Functional analysis has to be performed to investigate the leukemogenic potential of these residual cells.     

Chromosome change at 16q22 in nonlymphocytic leukemia: clinical implication on leukemia patients with inv(16) versus del(16)

Nineteen patients with inv(16)(p13q22) or del(16) in myeloid leukemia are described. Eight showed inv(16)(p13q22), including one with de novo acute myeloid leukemia (AML-M2) and seven with de novo acute myelomonocytic leukemia (AMML-M4). Additional chromosome changes were detected in five of the cases; the most common change was trisomy 22. All but one of the de novo M2 and M4 leukemia patients with inv(16)(p13q22) showed initial bone marrow eosinophilia (greater than 5%) with basophilic granules. The remaining 11 showed deletion of the long arm of a chromosome no. 16 [del(16)(q22 or q23)]. Eight of the 11 were diagnosed as having chronic myelomonocytic leukemia, three transformed into an acute phase with M4 morphology; none of them gained complete remission. Two of the remaining three patients with del(16) were diagnosed as having M4 leukemia without marrow eosinophilia. The remaining one was a case of M4 leukemia following a myelodysplastic syndrome. The findings indicate that del(16) might be related to chronic myelomonocytic leukemia or leukemia with a prior history of myelodysplastic syndrome without evidence of marrow eosinophilia. On the other hand, inv(16)(p13q22) is highly associated with de novo AML especially AMML-M4 with bone marrow eosinophilia and a favorable prognosis.     

Abnormalities of chromosome 16q in myeloid malignancy: 14 new cases and a review of the literature.

Fourteen patients with abnormalities of chromosome 16q, 13 with acute myelogenous leukaemia (AML), and one with refractory anaemia with excess of blasts (RAEB), are described. Seven patients had inv(16)(p13q22), two had del(16)(q22), and five had other abnormalities of 16q. Six of the seven patients with inv(16) had AML M4Eo and, following treatment with adriamycin, cytosine arabinoside, and 6-thioguanine, all achieved complete remission (CR). Neither patient with del(16)(q22) had typical M4Eo morphology at diagnosis; CR was achieved in one and one had resistant leukaemia. Patients with other abnormalities of 16q had blasts of diverse morphology and, although morphologically abnormal eosinophils were seen in three patients, this was not as marked as in the patients with inv(16). CR was achieved in two of the four patients with other abnormalities of 16q but duration of remission was short in both cases. These results suggest that most patients with del(16)(q22) and other abnormalities of 16q22 do not have typical AML M4Eo. Such patients tend to have a worse prognosis, and are more likely to have complex karyotypes typical of secondary leukaemia.     

"Acute myelogenous leukemia like" translocations in CML blast crisis: two new cases of inv(16)/t(16;16) and a review of the literature

We describe two patients with CML blast crisis with clonal evolution affecting 16q22 (t(16;16)(p13;q22) and inv(16)(p13;q22), abnormalities of core binding factor, usually found in de novo acute myeloid leukemia (AML)). The bone marrow of both cases showed myelomonocytic (M4) differentiation and eosinophilia. Both patients had prominent extramedullary disease and had poor response to treatment. A literature search focused on patients with CML and additional chromosome changes more typical of AML, revealed that the morphology of the blasts correlated with the finding typical of the underlying "AML" cytogenetic abnormality and an overall very poor clinical outcome, even in the groups with "favorable" AML type translocations.     

成人急性髓系白血病的免疫学特点及预后

目的：探讨急性髓细胞白血病（AML）的免疫表型特点。方法：使用淋系和髓系单抗,用间接免疫荧光法对70例原发性AML进行免疫表型分析。结果：所有AML患者的细胞至少被1种髓系单抗标记,各髓系抗原的表达率依次为CD33＞CD13＞CD15。所有M3患者CD9为阳性。16／70例（30％）表达CD34抗原、CD34^ AML组在年龄、外周血象及骨髓原始、幼稚细胞比例等方面与CD34^-组相比较无显著差别,但表达CD34抗原的AML常伴有HLA－DR、CD38、CD7等不成熟细胞表面标记的表达,而较成熟的髓系细胞表面标记CD15则不表达。70例AML中有16例表达淋系抗原,CD4^ 例（13．8％,M2为8．8％,M460％）;CD7^ 9例（16．9％,M1为50％,M218％,M5b16．7％）。CD4^ 的AML患者CD34为低表达,CD33表达。结论：CD9^ 、CD34^-、HLA－DR^-及CD13^ 、CD15^ 是典型M3的免疫表型特点。CD34^ AMLgn AML-M1有着密切的关系,且对化疗反应较差,证明CD34^ 的AML是一组分化程度较差的类型;提示CD7^ 和CD4^ 的AML预后差,CD7^ 的AML的一种独特类型。     

Extramedullary infiltrates of AML are associated with CD56 expression, 11q23 abnormalities and inferior clinical outcome

We evaluated the frequency and prognostic significance of extramedullary infiltrates (EMI) at presentation of acute myeloid leukemia (AML) in adult patients. Of 331 cases with de novo AML, 101(30.5%) had extramedullary infiltrates at diagnosis. The extramedullary manifestations included: lymphadenopathy, splenomegaly, hepatomegaly, gingival hypertrophy, skin infiltrates and involvement of central nervous system (CNS). Patients with EMI had a high initial WBC count and a high proportion of M4/M5 morphological variants. The complete remission rate (CR) with induction chemotherapy was lower in patients with EMI (P=0.0077) and their overall survival was also inferior (P=0.0017). Flow cytometric evaluation of the surface antigens expressed by the leukemic blasts for CD34, TdT, HLADR, CD7, CD19 and CD56 found that only CD56 expression was associated with EMI. The association of CD56 expression with lymphadenopathy was statistically significant (P=0.035). Abnormal karyotypes were found in 50.6% of patients with EMI and 49.7% of patients without EMI. Only 11q23 abnormalities were associated with specific sites of EMI; lymphadenopathy (P=0.0111) and gingival hypertrophy (P=0.0016). Our study of adult AML patients demonstrates that EMI at diagnosis is associated with CD56 expression by leukemic blasts, 11q23 karyotypic abnormalities, low complete remission rate and poor overall survival.     

Prognostic importance of cytogenetic subgroups in de novo pediatric acute nonlymphocytic leukemia

Reports of close associations between recurring chromosomal abnormalities and the clinical behavior of acute nonlymphocytic leukemia (ANLL) have stimulated efforts to define this disease in cytogenetic terms. Here we report on the leukemic cell karyotypes of 155 children with ANLL who were treated from 1980 to 1987 in consecutive programs of chemotherapy at this institution. Of 121 cases with adequate banding, 20% were normal, 30% had miscellaneous clonal abnormalities, and 50% were classified into known cytogenetic subgroups: inv(16)/del(16q) (n = 15), t(8; 21) (n = 14), t(15;17) (n = 9), t(9;11) (n = 9), t(11;V)/del(11q) (n = 7) and -7/del(7q) (n = 6). The inv(16)/del(16q) cases showed a nearly equal distribution of myelocytic and monocytic French-American-British (FAB) subtypes; only four of these patients presented with M4Eo morphology. Despite a 100% remission induction rate, patients with inv(16)/del(16q)-positive ANLL fared no better overall than the entire group; only 40% of this subgroup were event-free survivors at 2 years from diagnosis (P = .23). Patients with inv(16)/del(16q) frequently had CNS involvement at diagnosis (eight of 15) or initially relapsed in this site (three of eight). Event-free survival (EFS) was clearly superior for young patients with FAB M5 leukemia and the t(9;11) (P = .041). These patients were clinically indistinguishable from others with the FAB disease subtype, yet their responses to etoposide-containing therapies were noteworthy. By contrast, children with structural abnormalities involving 11q23, other than t(9;11), were infants (median age, 6 months) with FAB M4 or M5 leukemia, hyperleukocytosis, and frequent coagulation abnormalities. Patients with such changes [t(11;V) or del(11q)] relapsed early during postremission therapy: none remained disease-free more than 16 months from diagnosis. Because of resistant leukemia, patients with monosomy 7/del(7q) had a poor remission induction rate (17%; P = .0015); patients with the t(15;17) were also poor responders to induction therapy (44%; P = 0.02) because of hemorrhagic deaths. These results identify several cytogenetic subtypes of pediatric ANLL that may represent unique disease processes for which more effective early cytoreduction [-7/del(7q), t(11;V)], better supportive care measures [t(15;17)], or more effective CNS prophylaxis [inv(16)/del(16q)] would be warranted.