原发性浆细胞白血病的诊断和治疗

原发性浆细胞白血病(primary plasma cell leukemia,PPCL)是一种罕见的、具有高度侵袭性的浆细胞疾病,对常规治疗反应率低且预后不良.PPCL发病前无浆细胞骨髓瘤病史.近年来,随着新型药物如硼替佐米以及新治疗方法如造血干细胞移植的应用,PPCL患者的预后和生存得到了改善.本文对PPCL诊断和治疗的最新进展进行综述,以期为临床实践提供有益的借鉴.     

Thirty patients with primary plasma cell leukemia: a single center experience

The primary plasma cell leukemia (PCL) is a rare aggressive plasma cell dyscrasia. We investigated its clinical and laboratory aspects in a large series of patients. Among 934 consecutive patients with multiple myeloma (MM), registered between 1978 and 2004 in a single institution, 30 patients [M/F: 22/8; median age (yr): 60, range: 36-79] with PCL (3.1%) were diagnosed. Retrospective analysis of the clinical, immunophenotypic, and cytogenetic aspects was performed. All patients had anemia, thrombocytopenia, circulating plasma cells (median count 4 x 10(9)/l), and in 18/30 patients hypercalcemia was found. Extramedullar involvement was present in 18/30 (60%) patients. The plasma cells were CD138+ and CD38+ (9/9), CD20+ (1/9), and CD10+ (1/9) with cytoplasmic positivity for light chains (9/9). The cytogenetic studies, evaluable in 21/30 patients, showed normal karyotype (6/21), complex hypodiploidy (6/15), pseudodiploidy (5/15), and hyperdiploidy (4/15). Treatment modalities had no impact on survival (median 4.5 months). Seven patients achieved remission. The performance status (ECOG >or= 2), platelet count or= 460 U/l were independent prognostic parameters of survival. The immunologic and cytogenetic presentation of patients with PCL bears little significance on prognosis, which is heavily compromised by advanced age at diagnosis and the poor performance status.     

Flow analysis of hairy cell leukemia

Cellular DNA content, Coulter volume and light scatter were measured in cell suspensions from spleens or peripheral blood in 6 patients with hairy cell leukemia. In 2 of 6 cases, the DNA distribution obtained by flow microfluorometry demonstrated an abnormal cellular DNA content of the G₀?G₁ cells. This abnormality was confirmed by mixing the patients' cells with normal blood mononuclear cells. This is the first report of cellular DNA abnormalities in hairy cell leukemia demonstrated by flow microfluorometry. A relatively low number of cells were observed in the S phase of the cell cycle in all cases. On Coulter analysis, “hairy” cells displayed a modal volume 2 to 3 times larger than that of non-malignant lymphoid populations. Differences between “hairy” cell populations and normal lymphoid cells were also noted using light scatter measurements of ethanol-fixed cells, but they were less marked than those observed by Coulter analysis.     

112例淋巴系统恶性肿瘤骨髓免疫表型分析

背景与目的:淋巴细胞白血病和淋巴瘤骨髓侵犯的诊断以细胞形态学为基础,而免疫分型可通过获得肿瘤细胞分化和发育阶段的信息使淋巴系统恶性肿瘤的诊断更为准确,为临床合理治疗和预后判断提供重要的科学依据.本研究应用多参数流式细胞术(flow cytometry,FCM)探讨淋巴细胞白血病和非霍奇金淋巴瘤(non-Hodgkin's lymphoma,NHL)骨髓侵犯的免疫表型特点.方法:收集112例病理确诊NHL并伴骨髓侵犯和淋巴细胞白血病患者的骨髓标本.应用FCM检测肿瘤细胞的免疫表型.结果:45例前驱B淋巴母细胞白血病/淋巴瘤(precursor B lymphoblastic lymphoma/leukemia,B-ALL/LBL)主要表达CD19、CD10、TdT、CD34、HLA-DR和CD20;32例前驱T淋巴母细胞白血病/淋巴瘤(precursor T lymphoblastic lymphoma/leukemia,T-ALL/LBL)主要表达胞内CD3(cytoplasmic CD3,CyCD3)、CD7、CD5、TdT、膜表面CD3(surface CD3,sCD3)和HLA-DR.77例前驱淋巴细胞肿瘤中,28例(36%)有髓系抗原CD13、CD33的表达;9例(20%)B-ALL/LBL病例有CD20与CD34共同表达,28例(87.5%)T-ALL/LBL病例有CyCD3与TdT共同表达.成熟淋巴细胞肿瘤35例,其中17例慢性淋巴细胞白血病/小淋巴细胞淋巴瘤主要表达CD19、CD20、CD5和HLA-DR,并有CD19与CD5共同表达.4例弥漫大B细胞性淋巴瘤主要表达CD19、CD20、CD10和HLA-DR.3例伯基特淋巴瘤主要表达CD19、CD10、CD20、SIgM.1例套细胞淋巴瘤表达CD5、CD19、CD20、HLA-DR.5例外周T细胞淋巴瘤(PTCL)主要表达sCD3、CD5、CD7、CD4或CD8.1例间变性大细胞淋巴瘤主要表达sCD3、HLA-DR.4例NK/T细胞肿瘤表达CD56、HLA-DR,也表达CD7或CD4或CD8.成熟淋巴细胞肿瘤不表达早期抗原如CD34、TdT.成熟淋巴细胞肿瘤可伴有髓系抗原CD13、CD33的表达.结论:淋巴系统恶性肿瘤侵犯骨髓采用形态学结合FCM免疫学分型可获得T、B细胞来源、肿瘤细胞分化阶段和异常抗原表达等参数,有助于临床诊断和微小残留病灶的检测.     

Successful treatment of multi-agent chemotherapy with rituximab for IgM plasma cell leukemia

A 67-year-old woman presented with impaired general performance, suffering from fatigue, dyspnea on exertion, and paresthesia of the finger tips. The laboratory findings showed increased white blood cells at 11.37 × 10³ cells/μl with 26.5% abnormal cells, low haemoglobin and, elevated creatinine, although serum lactate dehydrogenase and calcium levels were normal. Serum immunofixation was positive for monoclonal IgM-kappa paraprotein. Total serum protein and the IgM component were elevated. X-ray examination of the skeleton was normal. Bone marrow aspiration showed 59.5% infiltration of abnormal cells that were characterized by typical mature plasmacytoid morphology. Abnormal cells expressed surface CD20, surface CD138, and cytoplasmic IgM, but not surface CD56 nor surface IgM by flow cytometric immunophenotyping with CD38 gating. Immunohistochemistry showed surface CD38, surface CD20, and cytoplasmic IgM. The clinical findings led to the diagnosis of the IgM Plasma cell leukemia (PCL). The patient recived multi-agent chemotherapy (VAD and EDAP with rituximab). The clinical symptoms disappeared, leading to the tumor load reduction.

To the best of our knowledge, this is the first report of successful treatment of multi-agent chemotherapy with rituximab for IgM PCL.     

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。     

B细胞幼淋巴细胞白血病七例免疫表型分析

目的 探讨B细胞幼淋巴细胞白血病(B-PLL)免疫表型特征.方法 通过多参数流式细胞术检测并回顾性分析7例B-PLL患者免疫表型,包括CD5、CD10、CD19、CD20、CD22、CD23、CD25、CD27、CD38、CD45、CD148、CD200、ZAP-70、FMC-7、κ、λ.结果 所有患者泛B细胞标志CD19、CD20、CD22均为阳性,其中仅有1例CD20呈中等强度表达,其余6例CD20均呈强阳性表达,CD22仅1例表达较弱,其余6例均呈中等强度表达.所有患者CD10均为阴性,4例CD5阳性,6例CD23阳性,2例FMC-7阳性,1例CD38阳性,4例CD200阳性,CD148均为阳性,轻链限制性表达检测中3例κ型,4例λ型.根据Matutes慢性淋巴细胞白血病免疫表型积分系统评分,1分1例,2、3分各3例.结论 B-PLL的诊断需要整合临床表现、外周血细胞形态学、免疫表型、细胞遗传学、分子生物学等结果综合判断,CD148、CD200或许可以成为B-PLL辅助诊断标志,B-PLL的免疫表型分布特征仍需要增加样本量进一步探讨.     

外周血干细胞CD+34/CD-38亚群与集落形成能力的相关性分析

目的探讨外周血干细胞亚群CD34+/CD38-与CFU-GM,BFU-E的相关性及其影响因素.方法应用流式细胞仪测定外周血干细胞冻存前后的CD34+及CD34+/CD38-,对冻存前后的PBSC进行CFU-GM,BFU-E测定.对数据进行对比分析及相关性分析.结果冻存后较冻存前集落产率明显降低(P＜0.05),而冻存1个月与3个月的集落产率差异无显著性(P＞0.05).冻存前后CD34+/CD38-百分率与CFU-GM、BFU-E的数目相关性分析显示二者均存在显著相关性.结论外周血干细胞CD34+/CD38-百分率与CFU-GM,BFU-E集落产率之间存在显著相关性,二者可作为测定PBSC质与量的互补指标.冷冻保存时间对PBSC的膜分子抗原无明显影响.冷冻保存减少了CFU-GM,BFU-E的产率,但冻存1个月与3个月对PBSC的集落产率影响不明显.     

Flow cytometric DNA analysis of interleukin-2 responsive renal cell carcinoma

Adoptive immunotherapy using interleukin-2 (IL-2) based therapy can result in marked tumor regression in some patients with metastatic renal cell carcinoma. DNA flow cytometry has not been previously studied as a predictor of outcome of this therapy. Archival paraffin embedded tumors were studied in 23 IL-2 treated patients with metastatic renal cell carcinoma. Eleven patients were complete responders (CR) and 12 were nonresponders (NR). In the CR group, 4/11 (40%) were diploid and 7/11 (60%) were aneuploid. In the NR group, 9/12 (75%) were diploid and 3/12 (25%) were aneuploid. Although there was a trend that patients with an aneuploid DNA pattern were more likely to undergo a complete response, ploidy pattern alone was not significantly predictive of response (p² = 0.10, Fischer's exact test). When combining ploidy pattern with other variables that were predictive for complete response, such as good performance status and a higher pretreatment weight, prediction of complete response was not improved by including ploidy. This preliminary report suggests that DNA ploidy does not appear to provide any additional information concerning responsiveness to IL-2 based immunotherapy beyond that obtained by performance status and pretreatment weight in this patient population. © 1993 Wiley-Liss, Inc.     

癌旁组织增殖特性的流式细胞测量术分析

 作者用流式细胞测量术对304例常见恶性肿瘤患者癌旁组织增殖特性进行了研究。结果表明，癌旁组织S期细胞比率(SPF)和超二倍体细胞比率(>2C%)均显著高于正常对照组织(P<0.01)，与癌中心灶十分接近。癌旁组织的DNA倍体类型不同，其>2C%明显不同，异倍体者显著高于二倍体(P<0.05)；癌旁组织倍体类型对其SPF影响不明显。癌中心灶DNA倍体类型对癌旁组织的>2C%和SPF未见明显影响。随肿瘤体积增大，癌旁组织的SPF有逐渐减低的倾向，其中肝癌旁组织表现最明显(R=-0.968，R>R0.01)；随肿瘤增大，癌旁组织超四倍体细胞比率(>4C%)呈递增的变化(R=0.84，R>R0.05)。     

急性粒-单核细胞白血病的免疫表型及细胞遗传学特征

背景与目的:白血病中某些抗原的表达与细胞遗传学的改变密切相关。本研究旨在探讨急性粒-单核细胞白血病(acutemyelomonocyticleukemia,M4)的免疫表型特征,并分析其与细胞遗传学的关系,为其诊断及治疗提供依据。方法:采用一组系列相关单克隆抗体和三色流式细胞术对81例M4患者进行免疫表型分析,并分析有核型资料的73例患者的核型异常;同时应用双色间期荧光原位杂交(fluorescenceinsituhybridization,FISH)技术对有染色体标本的35例患者进行inv(16)检测,并分析inv(16)阳性患者的免疫表型特征。结果:81例M4患者中48例(59.3%)表达干细胞标志CD34;髓系标志以CD33(84.0%)表达最高,其次为CD13(81.5%)和CD14(24.7%);分别有23例(28.4%)和10例(12.3%)患者表达T系和B系相关抗原。11例M4Eo患者中CD13均阳性,10例CD33阳性,7例CD34阳性,5例CD2阳性。FISH检测发现14例患者inv(16)阳性,其中CD13阳性13例,CD33阳性11例,CD34阳性8例,CD14阳性5例,CD7阳性3例,CD2阳性3例。73例M4患者中,异常染色体检出率为41.1%(30/73),共检出11种主要异常核型。CD2和CD34在染色体异常患者中表达明显增高,而CD14表达减低。CD2在M4Eo中表达增高,但其表达与inv(16)无显著性相关。结论:急性粒-单核细胞白血病以髓系抗原表达为主,CD2和CD34在染色体异常患者中表达明显增高;CD2在M4Eo中表达增高。Inv(16)在M4中占40%,各抗原表达与inv(16)无显著相关性。     

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。     

CD(45)单抗在急性白血病免疫分型中的作用

目的：探讨CD(45)设门在急性白血病免疫分型中的可行性和实用价值。方法：采用CD(45)单抗直接免疫荧光标记和流式细胞仪对32名健康人外周血白细胞及56例急性白血病患者骨髓中的细胞进行了检测和分析。结果：淋巴细胞门内，正常对照组相对急性白血病患者组CD(45)-FITC阳性细胞的百分比和平均荧光道数值分别为(±s)：(99.1±0.5)％对(85.2±4.3)％(P＜0.05)和31.66±8.47对2.46±1.07(P＜0.001)；CD45-PE为(99.3±0.7)％对(90.2±6.6)％(P＜0.05)和106.99±25.94对13.86±9.51(P＜0.001)。急性白血病患者的原始幼稚细胞与外周血白细胞1：1000混合稀释后，CD(45)／SSC参数图上，可清楚地对原始幼稚细胞设门。结论：利用CD45设门方法，能简单、快速、准确地识别混在外周血中的少量原始幼稚细胞，敏感度高达10-3。     

双表型急性白血病流式细胞术免疫分型检测结果

 目的　应用流式细胞术（FCM）检测双表型急性白血病（BAL）的免疫表型。方法　采用四色FCM检测23例BAL患者的免疫表型。结果　23例BAL患者中有10例（43.4 %）表达cCD3,16例（69.6 %）表达cCD79a,20例（87.0 %）表达cMPO,14例（60.9 %）表达TdT,19例（82.6 %）表达CD34,20例（87.0 %）表达CD117,同时表达髓系和B系抗原者13例（56.5 %）,均共同表达cCD79a和cMPO;同时表达髓系和T 系抗原者7 例（30.4 %）,均共同表达cCD3和cMPO;同时表达T系和B系抗原者3例（13.04 %）,均共同表达cCD3和cCD79a。结论　cCD3、cCD79a、cMPO为系列特异性抗原标志,对诊断及鉴别BAL具有重要意义,FCM是目前诊断BAL特异可靠的方法,在临床白血病治疗和预后方面有重要的指导意义。     

流式细胞术对食管癌组织中四种细胞周期调控蛋白的定量检测及意义

目的 探讨细胞周期调控蛋白cyclin E、cyclin D1、CDK4和D27在食管癌的表达及其与食管癌的分化和淋巴结转移的关系。方法 采用流式细胞术对65例食管鳞状细胞癌组织中cyclin E、cyclin D1、CDK4和p27的表达强弱进仃定量检测,结果 用荧光指数F1表示。结果 cyclin E、cyclin D1和CDK4在低分化型鳞癌的表达量显著高于分化型鳞癌(P值分别为0．0275,0．0001和0．0174);而p27在低分化型鳞癌的表达量显著低于分化型鳞癌(P=0．0042)。cyclin D1与cyolin E,cyclin D1与CDK4之间呈显著正相关;而cyclin D1与p27呈显著负相关。4种基因蛋白的表达与淋巴结转移均无相关性。结论 cyclhi E、cyclin D1、CDK4和p27的表达与食管癌的分化密切相关;正负性细胞周期调控蚩白表达的失衡是导致癌变的重要原因之一。     

流式细胞仪动态检测成人急性白血病细胞动力学的临床意义

应用流式细胞仪（ＦＣＭ）动态检测３４例成人急性白血病患者１４６份骨髓标本的细胞动力学参数，并以１０例正常人骨髓标本为对照。结果显示：（１）成人急性白血病患者骨髓标本Ｓ％在不同临床阶段存在一定差异。（２）白血病患者完全缓解时Ｓ％与正常组差异不显著；而巩固强化可使Ｓ％一过性降低。（３）典型病例动态检测表明不同化疗方案对Ｓ％的影响不同。说明动态观察Ｓ％的变化可为诱导和巩固强化的化疗方案个体化提供客观的细胞动力学参数。     

多参数流式细胞术检测急性早幼粒细胞白血病微小残留病的研究

 目的 建立流式细胞术检测急性早幼粒细胞白血病（APL,AML-M3）微小残留病（MRD）的方法,初步探讨其临床价值。方法 采用一组四色荧光标记单克隆抗体组合（CD15-CD11b-CD33-CD45）,对40例初诊时经形态学及流式免疫分型诊断为M3的白血病患者及其治疗后12例骨髓标本进行检测,同时检测正常对照8例。结果　在初发M3,以CD45／SSC（侧向角散射）设门,95 % M3白血病细胞群表达CD-15 CD-11b CD+33。12例在第1疗程结束进行监测,CD45／SSC设门粒细胞群CD-15 CD-11b CD+33细胞分别为0,0.001 %,0.002 %,0.07 %,0.08 %,0.09 %,0.22 %,0.25 %,0.66 %,1.73 %,12.24 %,53.6 %和72.4 %。该方法的敏感度为10-4,远高于形态学检测法。而诱导分化后的白血病细胞首先表达CD+15和（或）CD+11b,并逐渐分化为CD++15 CD++11b CD+/-33成熟粒细胞。结论　四色荧光标记单克隆抗体组合（CD15-CD11b-CD33-CD45）,可作为流式细胞术检测M3的MRD的方法。     

Flow cytometric measurements of heat shock proteins

A fixation and immunofluorescence staining procedure for measurements of heat shock proteins (hsp) by flow cytometry is reported. Three fixatives were compared: 80% methanol at —20°C for 1 h, 70% ethanol at 0°C for 1 h, and 3% paraformaldehyde at 4°C for 1 h followed by 0·2% NP-40. Cells fixed with methanol showed strongest immunofluorescence and lowest nonspecific fluorescence. The level of hsp 70 as a function of time after heating followed the same kinetics as the development of thermotolerance reported by others. The level of hsp 70 increased with increasing heat dose up to a maximum heat dose, and above this heat dose a decrease in the level of hsp 70 was observed. Correlated measurements of the level of hsp 70 and DNA showed that hsp 70 was found in all phases of the cell cycle. The level of hsp 70 increased about two-fold in unheated cells throughout the cell cycle. The increase in G₂ + M cells compared with G₁ cells was lower in cells heated at 45°C for 20 min followed by incubation at 37°C for 24 h before fixation and staining, than in unheated cells. The results show that flow cytometry provides a rapid and quantitative technique for measuring hsp. Correlated measurements of hsp and other cellular parameters might also be obtained.     

Flow cytometric analysis of DNA ploidy in large cleaved cell lymphomas

The results of nuclear DNA content analysis of a series of 28 patients with large cleaved follicular centre cell non-Hodgkin's lymphomas (NHL) are reported. DNA aneuploidy was found in 11 (39 per cent) cases. The DNA indices of the DNA-aneuploid peaks ranged from 1.14 to 2.28. Seven (25 per cent) cases were tetraploid. DNA ploidy was not associated with prognosis. The percentage of S phase cells (SPF) ranged from 2.0 to 30.5 per cent (median 5.1 per cent). Lymphoma patients with SPF higher than 9.7 per cent had a worse survival rate than patients with lymphoma with less than 9.7 per cent S phase cells but the difference did not reach statistical significance. The results of the DNA ploidy and SPF were comparable to those of intermediate and high grade malignancy NHLs.