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。     

急性髓细胞白血病微分化型流式细胞术免疫分型分析

目的探讨流式细胞术（FCM）检测免疫表型在诊断急性髓细胞白血病微分化型（AML—M0）中的意义。方法采用多色FCM分析14例AML—M0病例的各相关抗原表达情况。结果14例AML—M0中,仅1例依据骨髓细胞形态学作出诊断,其余13例均依靠FCM作出明确诊断。在AML—M0中,髓系抗原CD33 14例（100％）表达阳性,CD13和CD117 9例（64％）表达阳性,CD34和HLA—DR12例（86％）表达阳性,一般成熟髓系相关抗原如CD16、CD10、CD14等均阴性,B和T淋巴细胞特异抗原如CD79a和CD3均为阴性,少数原始细胞表达细胞内髓过氧化物酶（MPO）、TdT。常常表达一些淋系但并不特异的抗原如CD7、CD2或CD19,但比淋巴细胞白血病表达的荧光强度弱。结论FCM免疫分型在AML—M0诊断中至关重要。     

急性髓系白血病细胞表面同时表现淋系抗原的临床及免疫学分析

目的 探讨淋系抗原在急性髓系白血病细胞上的表达及意义。方法 采用间接免疫荧光法检测182例急性髓系白血病细胞的免疫表型。根据FAB亚型和免疫标志将病例分为2组：伴淋系相关抗原的急性髓细胞性白血病(Ly^ AML),不伴淋系相关抗原的急性髓细胞性白血病(Ly^-AML)。结果 182例AML中有64例表达淋系抗原(33％), CD7^ 在AML中表达率为54．7％,M0中阳性率最高100％,M1次之78％。Ly^ AML组的白细胞、血小板数高于Ly^-AML,有显著性差异。Ly^ AML,组与Ly^-AML组的诱导缓解率及临床特征无显著性差异。Ly^ AML组和Ly^-AML组比较,平均缓解期较短。结论 CD7^ AML与AML-M0、M1有着密切的关系。可以看作是一个独特的临床亚型。Ly^ AML较之Ly^-AML,具有不同的临床特征和短CR期。Ly^ AML的出现可以作为危险因子中的一个因子来选择更合适的化疗也许是必要的。     

急性髓系白血病细胞异常免疫表型表达的研究

应用免疫酶标染色法检测了59例急性髓系白血病（AML）患者的白血病细胞免疫表型，结果表明CD2、CD5、CD7、CD10、CD19、CD22淋系抗原的表达率分别为16．9％（10／59）、119％（7／59）、16．9％（10／59）、15．3％（9／59）、102％（6／59）和6．8％（4／59）。进一步分析结果表明，在M3病例细胞中，CD2、CD10和CD19抗原表达阳性率明显高于M5组，而CD7抗原表达阳性率则明显低于M5组。结合临床，CD2、CD19阳性的AML病例对化疗治疗及应优于CD2、CD19阴性的AML病例；CD7阳性的AML病例的疗效与预后则比CD7阴性的AML病例差。提示部分AML病例的白血病细胞存在不同程度异常免疫表型的表达，且与疗效及预后有一定关系。     

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     

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。     

儿童急性髓系白血病免疫表型特点分析

目的:分析儿童急性髓系白血病(AML)免疫表型特征,探讨其临床意义.方法:采用四色流式细胞术CD45/SSC双参数散点图设门方法,对127例儿童AML患者幼稚细胞进行免疫表型检测,对抗原表达情况进行分析.结果:在127例儿童急性髓系白血病患者中,髓系特异性抗原CD33、CD13和CD117的表达最常见,分别达95.3%、90.6%、90.6%.造血干/祖细胞抗原CD34、HLA-DR、CD38阳性率分别达53.5%、71.6%和97.6%.有65.4%的病例伴有淋系抗原的表达,其中以CD56的表达最常见占38.6%,其次为CD7占21.3%.有淋系抗原表达(LymAg+)患者早期抗原CD34和HLA-DR抗原表达阳性率明显高于无淋系抗原表达(LymAg-)患者(P<0.05).结论:免疫分型对儿童AML的诊断和不同亚型鉴别具有重要意义.     

35例急性白血病免疫表型特征

 目的 分析急性白血病（AL）免疫表型诊断意义并对急性白血病各亚型CD抗原表达进行比较。方法 对35例AL患者进行形态学检查,并用流式细胞仪检测白血病细胞免疫表型。结果 35例AL患者中,形态学分型：急性髓细胞白血病（AML）M1 2例,M2 4例,M3 6例,M4 7例,M5 8例,不能分型AL 1例,急性混合细胞白血病1例,急性淋巴细胞白血病（ALL）L1 4例,L2 2例。免疫表型检查结果在AML各亚型中CD13、CD33、MPO均阳性,CD+34 18例,CD+14 5例。其中2例伴有CD+19,6例ALL患者中,CD19、CyCD79a均阳性,CD+10 4例,CD+20 2例,1例不能分型AL表达CD10、CD19、CD34、CyCD79a诊断为B-ALL,1例混合细胞白血病有淋系和髓系表达,故诊断为混合细胞白血病。结论 白血病免疫表型检测能使AL的诊断率提高,因此对形态无法鉴别的AL,应进一步进行免疫表型检测。     

MIC分型评价成人急性髓系白血病中的淋系抗原表达

目的：研究成人急性髓系白血病（acute myeloid leukemia，AML）中淋系抗原的表达及其与预后的关系。方法：对275例成人AML进行细胞形态学、流式细胞免疫表型分析和染色体细胞遗传学（MIC）分型实验研究并对其中的64例M2进行临床治疗结果观察研究。结果：①AML淋系抗原阳性率31．3％，其中t（8；21）者为48．3％，无t（8；21）者为26，5％（P〈0．001）。②CD19^＋在t（8；21）中占38．3％，无t（8；21）中占1．9％（P〈0．001）；而其他淋系抗原两组比较均为P〉0．05．③治疗结果（CR率）：t（8；21）和无t（8；21）M2分别为66．7％和41．9％，CD19^＋和CD19^＋ M2分别为76．5％和46．8%，CD19^＋／t（8；21）和CD1^＋9／无t（8；21）分别为80．0％和41．4％，以上均为P〈0．05。而CD19^＋／t（8；21）和CD19^＋／无t（8；21）分别为55．6％和50．0％（P〉0．05）。结论：成人AML中淋系抗原表达与核型密切相关，t（8；21）AML高表达CD19。t（8；21）和CD19都是M2预后好的标志之一。     

新疆地区450例急性白血病的免疫分型研究

 目的　研究新疆地区急性白血病（AL）患者免疫表型分布特点。方法　采用间接免疫荧光法对450例AL患者进行免疫表型分析。结果　106例急性淋巴细胞白血病（ALL）,334例急性髓系白血病（AML）,10例为FAB不能分类的急性白血病（UAL）;ALL中髓系抗原的表达15 %,AML中淋系抗原的表达25 %,表达最频繁的是CD7;研究了295例AL患者MPO mRNA基因表达,81例ALL中有1例表达MPO基因;所有髓系均不同程度地表达MPO基因,9例UAL有6例表达MPO基因;ALL免疫分型特点在汉族和维吾尔族（简称维族）中差异无统计学意义（P＞0.05）,在AML中,汉族髓系抗原的表达率依次为CD33＞CD13＞CD15,维族髓系抗原的表达率依次为CD15＞CD33＞CD14。结论　免疫表型的检测对AL更精确地诊断和分型有重要意义。联合分析AL形态学、细胞化学、免疫学及MPO mRNA表达等特点,对于AL的诊断和指导治疗均有重要意义。     

Role of blast cell immunophenotyping for the diagnosis and prognosis of acute myeloid leukemia.

Bone marrow blast cell antigen expression from 86 patients with de novo acute myeloid leukemias (AML) was studied and correlated with FAB classification and clinical outcome. Among a panel of 14 monoclonal antibodies routinely used for the diagnosis of acute leukemias we studied the expression of six antibodies (CD13, CD15, VIM2, CD33, CD14, CD34) of the granulomonocytic lineage and found that some of them were useful for diagnosis and/or prognosis. For FAB subclassification of AML, the CD13 or VIM2 antigen expression was of no benefit. Monocytic leukemias (M4 + M5PD + M5WD) more frequently expressed CD34 antigen (28/31) than granulocytic (M1 + M2 + M3) subtypes (33/53) (P < 0.01). Finally, the most striking differences were found with CD14 antigen expression: CD14 antigen was more frequently expressed in M4 + M5 leukemias (21/31) than in M1 + M2 + M3 subtypes (12/33) (P < 0.01). The mean percentage of CD14 positive blast cells was accordingly higher in monocytic leukemias than in granulocytic leukemias and the difference was highly significant (P < 0.0001). The CD15 antigen was more frequently expressed in differentiated leukemias (M2 + M3 + M4 + M5WD) (35/44) than in poorly differentiated forms (M1 + M5PD) (17/37) (P < 0.001). The statistical difference was higher when the mean percentage of CD15 positive blast cells were compared (P < 0.0003). Moreover these latter percentages were different in M1 and M2 subtypes (P < 0.003). The blast cell expression of CD13, CD14, CD15 or CD33 was not predictive of the length of CR or survival. Moreover, our results support previously published findings suggesting a longer overall survival duration for patients whose leukemic cells do not express the CD34 antigen (P < 0.01). We also confirm that patients with the more differentiated subtypes of AML (CD13-, CD34+) tend to survive longer than patients with the less differentiated subtypes of AML (CD13-, CD34+) (P < 0.001).     

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。     

Immunological classification of acute myeloblastic leukemias: relevance to patient outcome.

Immunophenotyping is a major tool to assign acute leukemia blast cells to the myeloid lineage. However, because of the large heterogeneity of myeloid-related lineages, no clinically relevant immunological classification of acute myeloblastic leukemia (AML) has been devised so far. To attempt at formulating such a classification, we analyzed the pattern of expression of selected antigens, on blast cells collected at AML diagnosis. Patients were eligible if they had a first diagnosis of de novo AML and a sufficient number of blast cells for proper immunophenotyping. The relative expression of CD7, CD13, CD14, CD15, CD33, CD34, CD35, CD36, CD65, CD117, and HLA-DR were analyzed by cytometry in a test series of 176 consecutive AML cases. Statistical tools of clusterization allowed to remove antigens with overlapping distribution, leading us to propose an AML classification that was validated in a second AML cohort of 733 patients. We identified five AML subsets (MA to ME) based on the expression of seven antigens within four groups (CD13/CD33/CD117, CD7, CD35/CD36, CD15).-MA and MB-AML have exclusively myeloid features with seldom extramedullary disease and rare expression of lymphoid antigens. No cases of acute promyelocytic leukemia (APL) were observed within MB AML. MC AML have either myeloid or erythroblastic features. MD AML have more frequently high WBC counts than other subsets, which were related to the expression of CD35/CD36 and CD14 and to monoblastic differentiation. ME AML lack CD13, CD33, and CD117 but display signs of terminal myeloid differentiation. Specific independent prognostic factors were related to poor overall survival in each immunological subset: CD34+ (P<3 x 10(-4)) in MA AML, CD7+ in MB AML, non-APL cases (P<0.03) in MC AML, CD34+ (P<0.002) and CD14+ (P<0.03) in MD AML, CD14+ in ME AML (P<0.01). The inclusion of seven key markers in the immunophenotyping of AML allows a stratification into clinically relevant subsets with individual prognostic factors, which should be considered to define high-risk AML populations.     

Reactivity pattern of 'myeloid monoclonal antibodies' with emphasis on MCS-2

The reactivity pattern of the murine monoclonal antibody (MoAb) MCS-2 was tested on a panel of 724 cases of leukemia-lymphoma. MCS-2 was positive in 178/185 (96%) cases of AML (FAB M1-3), 10/10 cases of AMMol/AMoL (FAB M4/5), 42/45 (93%) cases of CML, 1/1 case of CMoL, 37/38 (97%) cases of CML-myeloid blast crisis, 0/9 cases of CML-lymphoid blast crisis. No positive staining was seen in 238 cases of T-CLL, mycosis fungoides, Sèzary-syndrome, B-CLL, hairy cell leukemia, multiple myeloma and T- and B-lymphoma nor in 32 cases of B-ALL, Burkitt-lymphoma, Null-ALL and immature T-lymphoma. A positive expression was found in 8/110 cases of cALL, 1/6 cases of pre B-ALL and 1/35 cases of T-ALL. Fifteen other MoAbs (MCS-1, OKM1, My-1, Leu-M1, Leu-M3, CA-2-38, MY4, MY7, MY8, MY9, VIM-D2, VIM-D5, Mol, Mo2, 63D3) which are associated with the myelomonocytic cell lineages were tested by indirect immunofluorescence on 60 or more patients (62-149 cases). A wide variability in the frequency of positivity was seen for the panel of cases studied and for the blast cell populations per individual samples: 21-96% of the AML cases (FAB M1-3) and 31-100% of the AMMoL/AMoL cases (FAB M4/5) were positive for the various MoAbs. None of the analysed MoAbs stained only myelocytic or only monocytic leukemias, but a certain degree of preference for the monocytic variants was noted for Leu-M3, CA-2-38, MY4, VIM-D2, Mo2 and 63D3. The detection of MCS-2 on immature ALL blast cells might indicate a coexpression of lymphoid and myeloid markers on very immature cells, or an abnormal gene expression by malignant cells, or the identification of a so far undetected subclass of acute leukemias.     

71例急性白血病免疫表型特征分析及意义

目的:分析急性白血病(AL)免疫表型特点及其临床意义。方法:采用单克隆抗体直接免疫荧光标记法的流式细胞术,对71例AL进行免疫表型检测。结果:71例AL患者以系列专一型表达为主,同时亦存在抗原交叉表达、不表达特异性抗原及混合型等情况。AL患者CD34和HLA蛳DR表达分别占56.3 %和61.9 %,M3患者均不表达HLA蛳DR。My+ALL患者完全缓解(CR)率(60.0 %)低于My - ALL患者CR率(80.6 %),两组相比有显著性差异(P<0.05)。CD+34 ALL与CD蛳34 ALL患者缓解率基本相同;CD+34 AML患者CR率(58.3 %)明显低于CD蛳34 AML患者CR率(88.9 %),两组相比有显著性差异(P<0.05)。结论:白血病免疫表型检测结合FAB分型可以提高诊断的准确率,部分免疫表型特征对判断预后有一定的意义。     

The CD4 molecule belongs to the phenotypic repertoire of most cases of acute myeloid leukemia.

In the present study fresh leukemic cells obtained from 23 patients with acute myeloid leukemia (AML; FAB subtypes: three M1, five M2, two M3, five M4, eight M5) were investigated for the membrane expression of the CD4 molecule by cytofluorimetric analysis with an anti-CD4 monoclonal antibody (mAb). In 15 cases the presence of the CD4 mRNA was also investigated using Northern blot analysis. Membrane expression of the CD4 molecule was demonstrated in 19 out of 23 cases, and it was found to be weaker than in CD4+ lymphocytes and monocytes obtained from normal controls. Full-length CD4 mRNA was detected in 12 out of 15 (80%) cases, and AML cells positive for CD4 mRNA expression also expressed the CD4 antigen. Since the CD4 molecule expressed by T cells is associated with p56lck, a member of the src family of intracellular tyrosine kinases, we investigated whether the CD4 molecule expressed by myeloid blasts is also associated with a tyrosine kinase activity. In vitro kinase assays performed on anti-CD4 immunoprecipitates from lysates of myeloid leukemia cells from four CD4+ cases were negative for the presence of a tyrosine kinase activity. This finding was not due to the lack of expression of members of the src family since we were able to detect at least p60src and p59fyn in myeloid leukemia cells. According to our results, the CD4 molecule seems to belong to the phenotypic repertoire of most AML, irrespective of their FAB subtypes. However, in myeloid blasts this molecule is not associated with a tyrosine kinase activity as it occurs in T lymphocytes.     

139例急性髓系白血病免疫分型特点分析

目的探讨急性髓性白血病（AML）的免疫分型特点及意义。方法采用单克隆抗体和流式细胞仪检测AML的免疫表型。结果（1）139例AML病例中各种抗原的阳性表达率依次为为MPO（92.1%）,CD33（92.1%）,CD13(89.2%),其中53例AML伴淋巴系抗原表达,分别为CD19(20.9%),CD7(16.2%),CD2(7.2%),CD10(0.72%)。(2)CD14在M4、M5型AML中高表达。(3)干祖细胞分化抗原表达率依次为CD117（83.8%）〉HLA DR(80.3%)>CD34(67.6%),CD34阳性的完全缓解率（CR）分别明显低于CD34阴性组（P=0.034）。（4）CD7阳性患者CR明显低于其抗原表达阴性者（P=0.041）。结论白血病免疫分型能确诊某些特殊类型的白血病,对免疫分型的研究将有助于指导临床诊断、治疗及判断预后。