急性白血病的细胞化学和免疫分型

目的 探讨急性白血病的细胞化学和免疫分型标准。方法 对52例急性白血病进行多种细胞化学染色和单克隆抗体免疫分型,并与以FAB为基础的形态学分型比较。结果 氯硝酸酯酶在鉴别粒细胞系统、醋酸萘酯加氟化钠抑制试验和鉴别单核细胞系中有重要作用。其中2例形态学、细胞化学不能分型的白血病,显示髓系单克隆抗体阳性,并发现5例混合型白血病同时表达粒系和淋巴系的抗原标志。结论 提示细胞化学和免疫学分型能明显提高急性白血病形态学诊断的正确率。     

急性白血病的细胞化学和免疫分型

近年来急性白血病的诊断准确率随着细胞化学和免疫分型的进展,有很大提高。细胞化学与单克隆抗体(单抗)免疫分型,以及两者与白血病细胞形态学分型的关系亦为许多人所重视。我们应用6种细胞化学和一系列单抗对急性白血病分型,并与FAB协作组的形态学分型相比较,以期说明联合应用对急性白血病分型的重要性。将结果报告如下。     

细胞化学染色在急性粒-单核细胞白血病分型中的应用

急性髓细胞性白血病在细胞形态学分型中,将急性粒一单核细胞白血病分为M4型。这是一种影响双细胞系列的白血病病变,骨髓细胞学变化多样及复杂,往往需借助于细胞化学和单克隆抗体的免疫分型作出诊断。本文分析了48例急性粒一单核细胞性白血病的骨髓细胞学特征和细胞化学特点,以期对此类型白血病的骨髓细胞形态学变化有足够的认识,提高诊断的准确性。     

急性粒-单核细胞白血病(M4)分型中细胞化学的应用

急性髓细胞性白血病的细胞形态学分型中,急性粒-单核细胞白血病被划分为M4型。这是一种影响双细胞系列的白血病病变,骨髓细胞学变化多样旦复杂,往往需借助于细胞化学和单克隆抗体的免疫分型作出诊断。本文分析了48例急性粒-单核细胞性白血病的骨髓细胞学特征和细胞化学特点,以期对此类型白血病复杂的骨髓细胞形态学变化有足够的认识,增加诊断的准确性。1 材料与方法 1.1 材料来源 全部病例均为我院门诊和住院初治病人,其中男性26例,女性22例,年龄15～58岁,中位年龄36岁。 1.2 形态学检查 骨髓涂片以瑞氏染色,按FAB分型标准进行分型,油浸镜下计数200个细胞,以百分数表示。M4a是原始的早幼粒细胞增生为主的M4,原幼单和单核细     

急性双表型白血病免疫分型特征的研究

目的 研究急性双表型白血病（BAL）免疫表型特征。方法 68例BAL患者骨髓标本进行细胞形态学及细胞化学染色确定其FAB亚型，运用一组系列相关单克隆抗体和流式细胞仪及直接免疫荧光标记技术进行免疫分型。结果 BAL患者细胞形态学以急性髓性白血病（AML）M1、M2亚型多见，免疫表型以髓系、B淋系共表达常见，CD34在BAL中表达较高。FAB诊断为AML的BAL患者中，B淋系CD10、CD19共表达的患者较多，诊断为急性淋巴细胞白血病（ALL）的BAL患者中，髓系CD13、CD33共表达者较多。结论 BAL为起源于比较早期的造血细胞且具有独特的免疫表型特征。     

急性白血病30例MIC分型回顾性分析

目的研究已经明确诊断的急性白血病患者骨髓细胞学,免疫学,遗传学的特征。方法对30例急性白血病患者的形态学、细胞免疫学、细胞遗传学（MIC）检测结果进行综合回顾性分析。结果显示以形态学为基础的FAB分型确诊率70%,以单克隆抗体免疫标记技术检测白血病细胞分化抗原显示与形态学符合率为70%,应用24 h培养G显带染色体组型分析,核型异常检出率66.7%。结论骨髓细胞形态学分析仍然是急性白血病确诊的基础,细胞免疫学,细胞遗传学的联合应用可提高急性白血病的确诊率,减少误诊率,对微小残留病灶监测及临床治疗提供翔实的实验诊断资料。同时,建议完善分子生物学检测,分析白血病患者的融合基因表达。     

105例儿童白血病的免疫分型与形态结果分析

目的 研究儿童白血病骨髓细胞形态学及免疫分型之间的关系.方法 对本院2016年新收治的105例儿童白血病进行回顾性分析,总结其流行病学特点及白细胞抗原的表达情况.结果 (1)ALL、AML患儿男女比例分别为2.13∶1,1.44∶1;中位发病年龄分别为3岁,2岁.(2) 105例儿童白血病FAB合型:初诊急性髓系白血病(AML) 22例,急性淋巴细胞白血病(ALL) 72例,未分类急性白血病2例,慢性粒细胞白血病(CML)4例,幼年型粒-单核细胞白血病(JMML)2例,神经母细胞瘤(NB)骨髓浸润2例,淋巴瘤骨髓转移1例;72例急慢性白血病免疫分型:急性髓系白血病(AML)16例,急性B淋巴细胞白血病(B-ALL) 47例,急性T淋巴细胞白血病(T-ALL)4例,慢性粒细胞白血病(CML)2例,幼年型粒-单核细胞白血病(JMML)1例,淋巴瘤骨髓转移1例,未见明显异常1例.(3)免疫分型与形态学分型完全符合率为91.67％(66/72),部分符合率为2.78％(2/72),完全不符合率为5.55％(4/72).(4)CD19、CD34、CD10、HLA-DR在儿童急性B淋巴细胞白血病中表达高,分别为100.00％(47/47)、80.85％(38/47)、89.36％(42/47)、100.00％(47/47);CD20在儿童急性B淋巴细胞白血病中表达均为12.77％(6/47);而CD13和CD33在儿童急性B淋巴细胞白血病中的表达率均为21.28％(10/47).结论 免疫分型对儿童白血病的诊断有重要作用,但并不能完全取代形态学分析,同时弥补了骨髓常规检查的不足.     

急性非淋巴细胞白血病形态学分型的探讨（附1142例分析）

我们以 1986年 9月在天津白血病分类分型讨论会上通过的急性非淋巴细胞白血病 (ANL L)的形态学分型诊断标准 ,对1987～ 1998年我院 1142例初诊 ANL L进行分析 ,对形态学分型提出意见与建议。1　材料和方法收集我院 1987年 1月～ 1998年 12月确认 ANL L 1142例。所有病例其化疗前血片和骨髓片均经瑞氏染色 ,并常规进行 pox、热盐水溶解试验 ,PAS、α- NAE及 Na F抑制试验 ,墨汗吞噬试验等辅助诊断。如系 M7,经电镜或单克隆抗体检测证实 ;如系 M4,行特异性酯酶与非特异性酯酶复合染色。2　结　果经统计 1142例确认 ANL L中 ,M1 2 7…     

急性淋巴细胞白血病24例形态学与免疫学分型分析

目的 探讨急性淋巴细胞白血病（ALL）形态学与免疫学分型的关系,以提高白血病的诊断水平.方法 针对24例ALL患者行骨髓细胞形态学、骨髓细胞化学染色及免疫分型检测,比较2者在ALL分型上的异同.结果 24例ALL患者中,FAB分型L1 15例、L2 7例、L3 2例;WHO免疫分型T-ALL 7例、B-ALL 17例.结论 免疫学分型是诊断ALL的重要手段,是对形态学分型的补充和修正,它对ALL患者的诊断、治疗和判断预后具有重要的指导意义.     

31例急性淋巴细胞白血病免疫分型的特点及与形态学分型的比较

目的:探讨急性淋巴细胞白血病的CD45设门三色流式细胞术免疫分型特征与诊断价值。方法:采用流式细胞术三色免疫荧光直接标记技术检测白血病患者,根据抗体积分系统进行分型。结果:三染色标记法检测24例B细胞系列急性淋巴细胞白血病(B-ALL)高敏感性和高特异性抗原分别为CD19(100%)和CyCD79 a(95.83%);7例T细胞系列急性淋巴细胞白血病(T-ALL)高敏感性和高特异性抗原分别为CD7(100%)和CyCD3(100%);CD10(85%),HLA-DR(73%),CD34(62%)等非系列相关性早期分化抗原在急性B淋巴细胞白血病均有较高的表达频率。31例急性淋巴细胞白血病患者中,急性淋巴细胞白血病(ALL)患者除均表达淋系抗原外,同时有64.52%伴髓系抗原(CD13,CD33)的表达。结论:流式细胞仪三色免疫荧光直接标记法进行白血病免疫分型能区分T-ALL、B-ALL,并可与急性髓性白血病相区别,尤其急性髓性白血病(M1和M5)。     

FLT3 Antibody‐based therapy for leukemia

Technological advances in antibody generation and production have facilitated recent clinical and commercial success with antibody‐based cancer therapeutics. The class III receptor tyrosine kinase FLT3 is highly expressed on the blast cells in most cases of acute myelogenous leukemia (AML) and B‐cell acute lymphoblastic leukemia (ALL). Activating mutations of FLT3 are detected in approximately 37% AML patients. FLT3 expression in normal tissue is limited to myeloid and B‐cell precursor cells. Therefore, over‐expressed or mutated FLT3 is an attractive target for therapeutic intervention using monoclonal antibodies. This review will discuss recent progress in the development of anti‐FLT3 antibodies as well as their therapeutic potentials in the treatment of AML and other hematological malignancies. Drug Dev. Res. 67:495–500, 2006. © 2006 Wiley‐Liss, Inc.     

Effects of monoclonal antibodies raised against the common acute lymphoblastic leukemia antigen on endopeptidase-24.11 activity.

The common acute lymphoblastic leukemia antigen (CALLA, CD10) has been identified as neutral endopeptidase-24.11 (NEP), a mammalian ectoenzyme involved in the inactivation of regulatory peptides, such as the enkephalins and atrial natriuretic peptide. Twenty monoclonal antibodies directed against the human antigen, were tested for their ability to inhibit the enzymatic activity of the human and rat peptidases expressed by cell lines. Six anti-CALLA antibodies were found to inhibit 50% or more of the hydrolysis of D-Ala2-leucine enkephalin by the neutral endopeptidase present on the human leukemic cell line Reh6 and, to a lesser extent, the hydrolysis of atrial natriuretic peptide. This may indicate that their binding may affect regions of the active site more important for the dipeptidylcarboxypeptidase activity of the enzyme. Only four antibodies cross-reacted with the peptidase from the rat epithelial cell line Rat2, as shown by membrane immunofluorescence, and these also partially inhibited enzyme activity. No antibody was able to inhibit completely the activity of the human and rat enzymes and all the active antibodies appeared to behave as non-competitive inhibitors of substrate cleavage. These monoclonal antibodies could be used in mapping studies of NEP.     

Report of a case with chronic myelomonocytic leukemia: demonstration of leukemic monocytes lacking nonspecific esterase by flow cytometry using monoclonal antibodies

Chronic myelomonocytic leukemia (CMML) is a rare leukemia, which is now included in myelodysplastic syndromes. In a small number of patients with CMML, problems in the diagnosis have been reported, especially when atypical morphological features in both monocytic and granulocytic cells due to dysmyelopoiesis are prominent, or when cytochemical characteristics are lost in the leukemic cells. The case history of a sixty-seven year-old male patient with CMML is described. The diagnosis of CMML in the patient was supported by the following evidence: chronic course of his disease; increased monocyte-like cells without other cause; normocytic anemia; immature granulocytic cells with hypogranular feature and giant platelets were observed in the peripheral blood. The bone marrow showed myeloid hyperplasia. Serum muramidase and vitamin B12 levels were increased, while neutrophil alkaline phosphatase score was low in the peripheral blood. Ph' chromosome was negative. The monocyte-like cells completely lacked nonspecific esterase. However the cells were confirmed as monocytic cells by flow cytometry using monoclonal antibodies to monocytes (OKM5).     

抗乙酰胆碱酯酶单克隆抗体的产生

将SP 2/O-Ag 14/小鼠骨髓瘤细胞和用乙酰胆碱脂酶免疫的Balb/c小鼠脾细胞在PEG-1000作用下进行融合,获14株分泌抗乙酰胆碱酯酶单克隆抗体的杂交瘤细胞系。用酶联免疫吸附试验(ELISA)对其分泌的抗体滴度进行了检测,其免疫鼠腹水滴度最高可达1:2 560 000。对其中5株杂交瘤分泌的滴度很高的抗体做了特异性交叉免疫试验,仅与乙酰胆碱脂酶发生特异反应,与其它4种酶(胃蛋白酶、木瓜蛋白酶、链霉蛋白酶及胰蛋白酶)无交叉反应;对其中12株杂交瘤分泌的抗体还进行了免疫球蛋白类型鉴定及染色体检查。上述杂交瘤细胞经3～6个月稳定传代后冻存于液氮中1年多后复苏,其抗体水平未见下降。     

流式细胞仪在儿童急性B淋巴细胞性白血病免疫分型中的应用

目的研究流式细胞仪检测儿童急性B淋巴细胞性白血病（B—ALL）免疫分型的临床价值，并探讨与预后的关系。方法利用流式细胞仪对39例B—ALL患儿在初诊时进行免疫分型。CD45-SSC设门，在CD19、CD10、CD20和CD22中选择两个单克隆抗体组合，进行三色组合分析。髓系抗体选用CD33和CD13根据是否伴有髓系抗原表达分为两组，并随访预后。结果①CD19、CD22、CD10的阳性率都超过或接近90％，分别为CD19：94．87％，CD22：92．31%，CD10：87．18％，而CD20只有15．38％的阳性率；有33．33％的患儿检测到髓系抗原（My）标志，其中以CD^13^＋8例，CD33^＋5例；②共有6例患儿复发，两组经，精确概率法检验，P〉0．05，差异无统计学意义。结论①流式细胞仪可以用于绝大多数的B—ALL患儿免疫表型的检测；②伴有髓系表达不具有提示复发的意义。     

Monoclonal antibodies for the treatment of acute myeloid leukemia

Currently, patients with acute myeloid leukemia (AML) are treated with cytotoxic chemotherapy and hematopoietic stem cell transplantation (HSCT). With this approach, the majority of patients still die of their disease because of both treatment-related mortality and relapse. Recently, monoclonal antibodies and immunoconjugates have been developed which potentially may increase the efficacy of treatment and decrease morbidity and mortality by specifically targeting the malignant cell. Unconjugated monoclonal antibodies have shown only moderate activity. A second, more effective, approach involves antibody conjugation with radioactive particles or chemotherapeutic agents, such as, immunotoxins, targeted delivery of cell killing. The antigens CD33, CD45, and CD66, are three antigens to which monoclonal antibodies have been directed. Most experience has been with gemtuzumab ozogamicin (Mylotarg) which is an immunoconjugate of an anti-CD33 antibody chemically linked to a potent cytotoxic agent, calicheamicin. Gemtuzumab ozogamicin appears to be particularly active in patients with acute promyelocytic leukemia, possibly related to the high expression of the CD33 antigen on the cell surface. Although gemtuzumab ozogamicin has activity as a single agent, the most promising result may be seen when this agent is combined with conventional cytotoxic chemotherapy. Preliminary studies have suggested a high complete remission rate and randomized clinical trials are underway. A unique potential toxicity has been identified, namely venoocclusive disease or sinusoidal obstructive syndrome which may be problematic among patients who subsequently undergo HSCT. An additional strategy includes radiolabeled monoclonal antibodies to intensify the conditioning regimen prior to HSCT. The most promising results have been obtained with radiolabeled anti-CD45 antibodies.     

Effects of ceftriaxone on severe infectious complications in hematological disorders. Tohkai Research Group on Infections in Hematological Disorders

Ceftriaxone (CTRX), a new long acting antibiotic in the 3rd generation cephem group, was administered intravenously once or twice a day in daily doses of 1-6 g for at least 3 days to 86 patients with severe infections complicating hematopoietic disorders. Underlying diseases were acute leukemia in 41 cases, chronic leukemia in 3 cases, malignant lymphoma in 19 cases, myeloma in 7 cases and others. Most patients (55 cases) suffered from sepsis or suspected sepsis. As for efficacy rates classified by underlying diseases, the treatment was effective in 61.0% of patients with acute leukemia. As for efficacy rates classified by infections, the treatment was effective in 60.0% of patients with sepsis. No side effects were noted except rash in 2 patients. Abnormal hepatic functions were recognized in 3 patients but were not attributed to the agent in any case. The results indicate that CTRX is a safe and useful antibiotic for the treatment of severe infections accompanied by hematopoietic disorders.     

86例成人急性髓性白血病免疫表型分析

常规使用10种单抗，用流式细胞仪（FACS）或荧光显微镜（FM）间接免疫荧光标记法检测了86树成人急性髓性白血病（AML）患者骨髓细胞的免疫标志。结果显示，髓系标志中CD33、CD13诊断价值较高，而CD15、CD14阳性率低。31．4％AML有淋系抗原表达，无单抗标记AML占16．3％，47．6％的患者表达CD34.FACS检测较FM更敏感、特异。     

In vitro neutralization by monoclonal antibodies of alpha-bungarotoxin binding to acetylcholine receptor

In order to develop monoclonal antibodies that would neutralize binding of alpha-bungarotoxin to acetylcholine receptor in vitro, mice were hyperimmunized with native toxin. Frequent small doses of toxin were used. Hybridoma supernatants were screened by ELISA and six monoclonal antibodies isolated and tested. The anti-alpha-bungarotoxin monoclonal antibodies consisted of IgM, IgG1 or IgG2a antibodies. In an in vitro neutralization assay measuring the effect of the antibodies on the binding of iodinated alpha-bungarotoxin to BC3H1 and TE671 (mouse and human cell lines bearing acetylcholine receptor), three of the six monoclonal antibodies were able to neutralize toxin binding. These studies demonstrate the feasibility of using native toxin for the generation of hybridomas, and the potential of using in vitro neutralization assays to screen hybridomas for in vivo neutralization.     

急性白血病患儿感染细小病毒B19的血常规和临床体征分析

目的 通过检测新发现的急性白血病患儿和接受过化疗的急性白血病患儿体内细小病毒B19DNA及其血清中的特异性抗体,探讨细小病毒B19感染对患儿血常规和临床体征的影响.方法 将2011年1月-2013年1月住院的95名急性白血病患儿分为两组.第一组：50名接受过化疗的急性白血病患儿;第二组：45名新发现、未接受化疗的急性白血病患儿.两组采用PCR法检测细小病毒B19DNA,以ELISA法检测血清细小病毒B19-IgG和B19-IgM抗体.结果 ①第一组检测细小病毒B19病毒DNA阳性率为32.0％,第二组为46.7％,对照组为零.②第一组当前感染的患儿有显著的血红蛋白和中性粒细胞水平下降及淋巴细胞增多;第二组当前感染的患儿有显著的中性粒细胞减少、血小板减少以及淋巴细胞增多.③临床体征方面,两组当前感染患儿都有明显的肝脾、淋巴结肿大和发热.结论 不管有无接受过化疗,细小病毒B19感染都是急性白血病患儿血细胞减少的重要原因.此外,细小病毒B19的感染与显著的肝脾、淋巴结肿大有一定的相关性.