The expression and clinical significance of CD59 and FLAER in Chinese adult AML patients

BackgroundThe role of CD59 and fluorescently labeled aerolysin (FLAER) in acute myeloid leukemia (AML) remains unclear and requires further investigation. To explore the relationship between CD59, FLAER, and AML, we investigated CD59 and FLAER expression in AML and analyzed their relationship with clinical characteristics of AML patients.MethodsWe employed flow cytometry (FCM) to analyze CD59 and FLAER expression in 161 AML patients at Tianjin Medical University General Hospital and evaluated its association with sex, white blood cell (WBC) count, platelet (PLT) count, thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), D‐Dimer(D‐D), and lactate dehydrogenase (LDH), followed by analyzing its connection with disease progression and complete remission (CR).ResultsCD59 and FLAER deficiencies were identified in AML patients. Compared with CR group, non‐CR group patients revealed more CD59 and FLAER deficiency. Compared with non‐acute promyelocytic leukemia (M3) group, M3 group patients had more CD59 and FLAER deficiency. CD59⁻ level in primordial cells of M3 patients was positively correlated with primordial cell ratio (r = 0.660, p = 0.003). Additionally, we discovered that the decline in CD59 and FLAER levels might be linked to higher D‐D and LDH in AML patients. The difference was statistically significant (p < 0.05).ConclusionsWe demonstrated that the decline in CD59 and FLAER levels was associated with leukemia cell proliferation and abnormal coagulation function in AML, suggesting that they could serve as a predictor of AML coagulation dysfunction, particularly in M3.     

Phenotypic conversion of T lymphoblastic lymphoma to acute biphenotypic leukemia composed of lymphoblasts and myeloblasts. Molecular genetic evidence of the same clonal origin.

Acute biphenotypic leukemia composed of lymphoblasts and myeloblasts developed in a patient with T lymphoblastic lymphoma (T-LBL) who had an anterior mediastinal mass. A novel myeloid cell line, termed TK-1, has been established from his peripheral blood after the leukemic conversion. The identical rearranged pattern of T cell receptor gamma-chain gene was observed among the DNAs derived from lymph node cells in the lymphoma phase, the myeloid cell line TK-1, and the subclones with different karyotypes (TK-1B and TK-1D), which showed that myeloid cells had been derived from the T-LBL of the same patient. This finding demonstrates that phenotypic conversion occurs in the clonally propagating tumor cells and suggests that some hematopoietic cells retain the capacity to adopt either lineage.     

成功治疗儿童罕见髓系/自然杀伤细胞祖细胞急性白血病1例

本文总结儿童髓系/自然杀伤细胞祖细胞急性白血病(M/NKPAL)的治疗经验以提高对该病的认识。对1例罕见的3岁8个月女童M/NKPAL合并中枢神经系统白血病进行了确诊分析,并对其治疗经过及长期随访结果进行了总结。结果表明,女童M/NKPAL合并中枢神经系统浸润得到了确诊,其免疫表型特征为CD7,CD33,CD34,CD56和HLA-DR共表达,MPO阴性,其他NK细胞和T、B细胞分化抗原阴性,染色体核型有+8和12p-。采用柔红霉素+阿糖胞苷化疗后达完全缓解,随后应用急性髓系白血病的化疗方案巩固强化治疗5个疗程,化疗方案中均含有大剂量阿糖胞苷,期间共行腰穿及鞘内注射治疗10次。停止治疗后中枢神经系统白血病复发,腰穿及鞘内注射治疗9次后行颅脑放疗36 Gy,取得了长期生存。结论:M/NKPAL是一种罕见的白血病,有特异的免疫表型特征,应用含有大剂量阿糖胞苷的急性髓系白血病化疗方案可能取得较好疗效。     

115例急性髓细胞白血病多参数流式细胞术免疫表型分析

免疫表型检测已成为白血病诊断和分类的重要手段。为了解急性髓细胞白血病(AML)免疫表型特征，利用三色流式细胞术CIM5／SSC参数散点图设门方法，对115例AML患者幼稚细胞表面及胞浆内分化抗原进行分析：结合FAB分类，比较AML不同亚型中抗原表达的差异，并对其诊断价值加以探讨。结果显示：在AML患者中，CD33、CD38和CDl3的表达最常见，分别达94．8％、91．3％、89．6％。在淋系抗原中，CD7的表达最为常见(20．2％)，其次是CD19(16．5％)和CD2(15％)，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、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，结论：多参数流式细胞术是诊断AML的一项可靠技术，AML某些免疫表型特征与FAB分类具有明显相关性．     

CD117在白血病细胞上的表达分析

目的　探讨细胞表面分化抗原CD117在各型白血病细胞上的表达规律及其意义。方法　采用CD4 5/SSC双参数散点图设门方法进行三色流式细胞术细胞表面分化抗原分析。结果　急性髓系白血病 (AML)患者CD117表达率为 6 8%,慢性粒细胞白血病急变期 (CML BC)患者CD117表达率为 80 %,而在急性淋巴细胞白血病中表达率极低 ,仅为 2 %。在慢性淋巴细胞白血病、CML慢性期中阴性。在AML中CD117主要表达在M0 /M1( 72 %)、M2 ( 88%)、M4 ( 5 0 %)、M5a( 75 %)、M6( 10 0 %) ,但在M3 和M5b中表达率较低 (分别为 39%和 2 9%) ,在M3 中CD117的表达率高于CD34 及HLA DR。CD117与CD14 在AML中的表达呈负相关。结论　CD117有助于淋巴系与髓系白血病的鉴别 ,并有助于白血病克隆与正常细胞的区分。     

CDll7在白血病细胞上的表达分析

目的 探讨细胞表面分化抗原CD117在各型白血病细胞上的表达规律及其意义。方法 采用CD45／SSC双参数散点图设门方法进行三色流式细胞术细胞表面分化抗原分析。结果急性髓系白血病(AML)患CD117表达率为68％，慢性粒细胞白血病急变期(CML-BC)患CD117表达率为80％，而在急性淋巴细胞白血病中表达率极低，仅为2％。在慢性淋巴细胞白血病、CML慢性期中阴性。在AML中CD117主要表达在M0／M1(72％)、M2(88％)、M4(50％)、M5b(75％)、M6(100％)，但在M3和M5b中表达率较低(分别为39％和29％)，在从中CD1l7的表达率高于CD54及HLA—DR。CD117与CD14在AML中的表达呈负相关。结论 CD117有助于淋巴系与髓系白血病的鉴别，并有助于白血病克隆与正常细胞的区分。     

Acute appendicitis caused by acute myeloid leukemia

A case of appendiceal involvement by acute myeloid leukemia (AML) in an adult with recent history of AML transformed from myelodysplastic syndrome (MDS) was presented. Being aware of this rare presentation in particular in a patient with history of MDS and/or AML is important for prompt clinical diagnosis and management.     

急性髓系白血病细胞CD34+抗原表达及与预后的关系

为了检测初治急性髓系白血病（AML）细胞CD34^＋抗原的表达,并分析其与预后的关系,应用间接免疫荧光法检测了238例AML患者.结果表明：238例AML中CD34阳性表达者有92例,占38.7%.除M3无CD34阳性表达外,CD34^＋表达与FAB亚型M0、M1有关;CD34^＋组完全缓解率为32%,明显低于CD34^-纽的61%;除淋系抗原CD7在CD34^＋组表达明显增高外,其它淋系抗原表达在两组间无差异（P＜0.05）.结论：有CD34^＋表达的AML患者预后差、CR率低,检测CD34表达对判断AML疗效有一定价值.     

CD68因子在急性髓系白血病患者中的表达情况及其临床相关性研究

目的:分析急性髓系白血病(AML)患者骨髓及外周血血浆中CD68因子的表达及临床特点.方法:采用四色流式细胞术检测50例初诊AML患者、23例对照组(非血液系统恶性疾病)的骨髓原始细胞CD68的表达情况.采用ELISA法检测85例初诊期、29例缓解期AML患者与24例对照组外周血血浆中CD68的表达情况.分析CD68因...     

伴髓系抗原表达的急性淋巴细胞白血病MIC分型特征分析

目的探讨伴髓系抗原（My）表达的急性淋巴细胞白血病（My^＋ALL）MIC分型特征。方法按常规骨髓涂片及过氧化物酶染色法对120例初治ALL患者进行形态学分型，采用流式细胞术检测白血病细胞免疫学类型，用R显带技术分析染色体核型，对My^＋ALL及My^-ALL患者MIC分型进行分析比较。结果120例初治ALL患者中My^＋ALL66例（55％），其中My^＋B-ALL50例，占B-ALL的56．8％：My^＋T．ALL14例，占T．ALL的50％；My^＋T、B-ALL2例，占T、B-ALL的50％。66例My^＋ALL患者形态学分型有10例（15．1％）误诊为急性非淋巴细胞白血病（ANLL）；54例My-ALL无一例误诊为ANLL。My^＋ALL患者形态学分型与免疫分型不符率或形态学分型不明确病例明显高于My-ALL（P〈0．01）。My^＋ALL患者95．5％表达CDl3，81．8％表达CD33，77．3％同时表达CDl3及CD33．1．5％表达CDll7。CDl4、CDl5及MPO表达阴性。My^＋ALL患者CD34阳性表达率明显高于My-ALL（P〈0．01）。My^＋ALL及My^-ALL患者异常染色体核型的检出率分别为72．3％和66．7％（P〉0．05），My^＋B-ALL患者的t（9；22）或t（9；22）伴其他染色体核型异常的检出率显著高于My^-B-ALL（P〈0．01）。My^＋T-ALL及My^-T-ALL患者未见t（9；22）异常。My^＋ALL和My^-ALL患者CR率分别为83．9％和79．0％，差异无统计学意义（P〉0、05）。结论My^＋ALLMIC分型部分细胞形态学可显示有髓系细胞特征，易被误诊为急性髓系白血病。My^＋ALL患者CD34表达率明显高于My^-ALL。My^＋B-ALL的t（9；22）异常显著高于My^-B．ALL。My^＋ALL与My^-ALL缓解率差异无统计学意义。     

CD56在急性髓系白血病细胞上的表达及微小残留病分析

目的探讨CD56在非M3急性髓系白血病(AML)中的表达及在微小残留病(MRD)检测的意义。方法应用流式细胞术分析145例非AML-M3 AML患者免疫表型,以CD56、CD19、CD117、CD34、CD33为标志,CD45设门MRD分析。结果 145例AML患者中CD56阳性37例,占25.52%,CD19阳性17例,占11.72%,分布在M1、M2、M4、M5。CD56或/和CD19阳性的AML患者共49例,其中CD56和CD19同时阳性5例,均为M2亚型。P170蛋白阳性36例,占24.83%,其表达率与CD56呈显著正相关(P<0.01)。48例AML患者在诱导治疗结束和维持治疗第14、32、56周作MRD检测。单纯CD56+组MRD阳性率93.55%,复发率77.41%,与单纯CD19+组(MRD阳性率25.00%,复发率0)比较,差异有统计学意义(P均<0.01)。CD56+CD19+组MRD阳性率100.00%,复发率80.00%,与单纯CD56+组比较,差异无统计学意义(P>0.05),与单纯CD19+组比较,差异有统计学意义(P<0.01)。结论 CD56、CD19在非M3 AML有较高的...     

Pretreatment prognostic factors for overall survival in primary resistant acute myeloid leukemia

Aim

Primary resistant acute myeloid leukemia has a very poor prognosis. We assessed pretreatment parameters for their significance as prognostic factors in the overall survival (OS) of 53 acute myeloid leukemia (AML) patients who had failed to achieve complete remission (CR) after first-line standard-dose remission-induction therapy.

Results

During the period January 2005–December 2009, 53 with acute myeloid leukemia received two cycles of the 3 + 7 protocol as a first-line standard-dose remission-induction therapy (ARA-C, days 1–7 and daunorubicin, days 1–3). The HiDAC (5 patients), MiDAC (7 patients), and FLAG-IDA protocols (3 patients) were given as salvage therapy. None of these patients achieved CR. There were 27 (51%) males and 26 (49%) females (median age, 55 years, range 28–76). The median white blood cell count was 53 (range 0.9 –350) × 10⁹/L, platelets 44 (range 3–856 × 10⁹/l) and bone marrow blasts 67%. HCT-IC comorbidity scores were 3 in two (3.8%) patients, 2 in 11 (20.8%), 1 in 12 (22.6%) and 0 in 16 (30.2%) patients. Median OS was 3.9 months (range 1 –20 months). The hepatomegaly, white blood cell count, ECOG PS, serum level of lactate dehydrogenase, dysplastic changes, coexpression of CD64, CD15, CD11b, comorbidities and disease cytogenetics influenced survival.

Conclusion

This single-center study evaluated the significance of pretreatment factors, and found that patient age, comorbidities, ECOG performance status, leukocytosis, hepatomegaly, LDH, and the disease cytogenetics were factors which influenced the outcomes of primary resistant patients with acute myeloid leukemia. An understanding of these factors may help to predict OS in cases where CR has not been achieved and may help when making further treatment decisions.     

免疫组化结果误判导致淋巴瘤误诊三例分析

目的 提高对正确分析判断免疫组化结果的重要性的认识.方法 对3例淋巴瘤误诊病例进行复习,并增加相关抗体标记予以鉴别诊断.结果 例1为淋巴结经典霍奇金淋巴瘤,富淋巴细胞型(LRCHL),误诊为滤泡性淋巴瘤与形态学观察遗漏R-S样细胞以及误判免疫组化标志BCL-2、CD20有关,即将BCL-2、CD20阳性的非肿瘤细胞误判为肿瘤细胞;误诊为结节性淋巴细胞为主型霍奇金淋巴瘤与免疫组化标志误判有关,把围绕瘤细胞的背景小淋巴细胞CD20阳性误判为R-S样大细胞CD20阳性,将CD30阳性的肿瘤性大细胞误判为活化性B淋巴细胞.例2为急性髓系白血病,误诊为T淋巴母细胞淋巴瘤(T-LBL),主要由于对瘤细胞表达非特异性抗体TDT、CD7、CD43的意义认识不足.例3为胸腺瘤B1型误诊为T-LBL,主要与抗体CK标记不理想而遗漏了肿瘤性上皮细胞,将表达TDT、CD99的反应性细胞误认为肿瘤细胞,以及对Ki-67表达率的意义认识不足有关.结论 淋巴瘤的诊断建立在形态学、免疫组化标志、临床资料和遗传学分析基础上,免疫组化结果的正确分析判断对淋巴瘤确诊至关重要.     

急性髓样白血病干细胞CD200的表达及与疗效的相关分析

目的探究急性髓样白血病（AML）患者白血病干细胞（LSCs）中CD200的表达以及其与临床疗效的相关性。 方法收集2014年4月至2017年4月衢州市人民医院收治的137例AML患者,进行AML分型,分为M1~M5型。并对患者临床疗效进行统计,分为三组：完全缓解（CR）组（59例）、部分缓解（PR）组（47例）和未缓解（NR）组（31例）。通过流式细胞仪分离患者治疗前后的骨髓LSCs,检测LSCs表达、CD200在LSCs中的表达水平。 结果24.82%（34/137）AML患者的LSCs中表达CD200,且CD200在AML各亚型中表达的阳性占比的比较,差异有统计学意义（5/11、13/26、4/35、3/22、9/43,χ²=16.533,P=0.002）。AML-LSCs在AML各亚型中都存在高表达,且差异无统计学意义（F=0.980,P=0.421）;除CD200在M3组的LSCs中呈现更低表达外[（10.1 ± 2.7）%],其在AML其他各亚型中表达差异均无统计学意义[（16.5 ± 2.8）%、（19.7 ± 4.0）%、（16.4 ± 2.4）%、（17.0 ± 3.1）%,P均> 0.05]。而在治疗疗效观察中,治疗前CD200在LSCs中的表达水平在CR、PR、NR三组间比较,差异均无统计学意义（P均> 0.05）;治疗后随着症状的缓解程度提高,各组CD200在LSCs中表达水平也随之降低[（16.4 ± 3.6）%、（10.8 ± 2.6）%、（5.0 ± 1.8）%,P均< 0.05]。治疗后,CR与NR两组CD200阳性表达占比的比较,差异有统计学意义（5/59 vs. 14/31,P < 0.017）;同时AML亚型分组中M3组内的CR、PR、NR三组CD200⁺/CD200^-分布情况的比较,差异有统计学意义（0/22、1/11、1/0,χ²=17.786,P < 0.001）。 结论AML患者LSCs中高表达CD200与不良预后有一定关系,可影响临床疗效。     

192例急性髓系白血病免疫表型、细胞遗传学及临床特征的研究

本研究对192例急性髓系白血病（AML）患者进行免疫表型检测,并探讨其与细胞遗传学改变和临床特征的关系。应用流式细胞术及一组系列相关单克隆抗体对192例AML患者骨髓进行免疫表型分析,用染色体G显带技术对其中的125例进行核型分析。结果显示,髓系抗原CD13、CD33、MPO、CD117表达最常见于AML。CD117表达于84．6％AML—M3病例中;较强的自发荧光、CD34与HLA—DR双阴性、表达相关髓系抗原CD13、CD33和MPO对于AML—M3诊断具有一定参考价值。CD14仅在AML—M4和AML—M5中表达;CD64和CD15同时强阳性伴HLA—DR高表达提示AML—M5可能性大。192例AML中,有47．9％伴淋系抗原表达,其中以CD7（20．8％）和CD56（26．0％）最常见,其次为CD19（9．9％）和CD2（7．3％）。125例AML中核型异常者76例（60．8％）。17例伴t（8;21）的AML—M2患者CD19、CD56、CD15表达均明显增加。另外28例t（15;17）均见于M3;2例inv（16）见于M4E0。LymAg^＋组CD34阳性患者比例（77．2％）明显高于LymAg-组（48．0％）。结论：免疫表型对AML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。本研究结果提示综合细胞形态学、遗传学及免疫表型分析,对AML患者诊断、分型及预后评估有重要意义。     

t(8;21)急性髓系白血病的免疫表型特点

目的研究用流式细胞术免疫表型分析t(8;21)(q22;q22)急性髓系白血病(AML)的免疫表型特点.方法采用常规细胞形态学/细胞化学、流式细胞术间接免疫荧光标记技术和R显带核型分析(MIC)进行分型,部分加用RT-PCR检测AML1/ETO融合基因.结果①294例初治AML患者中,64例t(8;21)AML,占21.8%,在M2中高达54.7%.64例t(8;21)AML中M2占81.3%;②与对照组比较,t(8;21)AML高表达CD19和CD34,低表达CD33(P＜0.001);③以20%阳性细胞作为阳性标准,CD19阳性率在AML为13.6%(294例中40例),t(8;21)AML为50%(64例中32例),显著高于无t(8;21)AML组的3.5%(230例中8例)(P＜0.001);④在t(8;21)AML患者中,CD19+和(或)CD34+58例(90.6%)、CD19-和(或)CD34-6例(9.4%).结论t(8;21)AML特别是M2/t(8;21)高表达CD19与CD34,CD19与t(8;21)密切相关,CD19是预测t(8;21)的指标之一.     

Optimization of the concentration of autologous serum for generation of leukemic dendritic cells from acute myeloid leukemic cells for clinical immunotherapy

Clinical application of immunotherapy for acute myeloid leukemia (AML) requires the efficient induction of dendritic cells (DCs) from AML blast cells using in vitro culture. We examined the effect of autologous serum on the properties of leukemic DCs derived from leukemic cells of AML patients by culture in AIM-V medium with GM-CSF, IL-4, TNF-alpha, and 0, 2, 5, or 10% human autologous serum. The expressions of CD80, CD83, CD86, and HLA-DR were upregulated under all culture conditions; however, 10% autologous serum induced the highest expression levels of several molecules. The capacity of leukemic DCs to stimulate allogeneic T cells increased with increasing serum concentration. Stimulation of autologous CD3(+) T cells with leukemic DCs grown in the presence of various concentrations of autologous serum resulted in induction of more IFN-gamma-secreting cells than was the case for unprimed CD3(+) T cells. Leukemic DCs cultured with 10% autologous serum induced the highest numbers of IFN-gamma-secreting cells and CD8(+)CD56(+) T cells from autologous T cells. These results suggest that culture of AML blast cells in the presence of autologous serum could be used to generate leukemic DCs for immunotherapy against AML. The highest serum concentration appeared optimal for generating the most potent leukemic DCs.     

急性髓系白血病患者NK细胞表达及功能变化研究

目的:探讨急性髓系白血病(AML)患者自然杀伤(NK)细胞的抗原表达特征和功能标记物的表达特点.方法:采用多参数流式细胞术检测56例初诊AML患者临床样本以及24例健康对照样本中的NK细胞表面标记物以及其功能指标,包括激活性受体NKG2D、NKP46、DNAM-1和杀伤标记颗粒酶B、穿孔素.结果:56例AML患者参照W...     

急性髓系白血病转换为急性淋巴细胞白血病1例并文献复习

目的 探讨急性髓系白血病(acute myeloid leukemia, AML)转换为急性淋巴细胞白血病(acute lymphoblastic leukemia, ALL)的临床特征,为该病的临床诊断及治疗提供依据。方法 回顾性分析1例AML-M5b转换为B-ALL患者的临床资料、诊疗经过并复习相关文献。 结果 患儿,女,4岁。发热伴腹痛5天。结合患者病史、体格检查及辅助检查,诊断为AML-M5b,诱导化疗后达到血液学及细胞遗传学的完全缓解(complete remission, CR)。1年后患儿白血病复发,形态学及免疫表型符合B-ALL。通过嵌合抗原受体T细胞(chimeric antigen receptor expressing T cells, CAR-T)治疗,患儿再次获得了 CR,但不久后疾病复发,二次CAR-T治疗无效,患儿死亡。结论 急性白血病复发后发生系别转换,预后较差,需要根据复发后表型作相应的治疗调整,若AML转换为B-ALL,表达CD19,CAR-T治疗可使其再次获得CR,但易二次复发。临床上,对于复发性白血病,需要完善白血病免疫分型,基因突变检测,肿瘤染色体等细胞遗传性及分子生物学等方面的检查,以便更好地指导治疗及评估预后。