30例慢性淋巴细胞白血病分子遗传学异常的FISH检测的意义

为探讨间期荧光原位杂交（FISH）在检测慢性淋巴细胞白血病（CLL）＋12、del（13q14）、p53基因缺失、atm基因缺失情况中的意义,采用组合探针（CEP12、LSI D13S319、LSI p53、LSI atm）对30例CLL患者进行FISH检测,分析分子遗传学异常与患者外周血淋巴细胞绝对计数、Binet分期、血清乳酸脱氢酶（LDH）和B：微球蛋白（β2-MG）水平、ZAP-70和CD38表达之间的相关性。结果发现,30例CLL患者中,19例（63．3％）存在1种及1种以上细胞遗传学异常,7例（23．3％）同时检测出2种及2种以上的异常,其中最常见为del（13q14）（43．3％）,其他依次为＋12（23．3％）,atm基因缺失（13．3％）,p53基因缺失（10．0％）。各分子遗传学异常与性别、年龄、Binet分期、外周血淋巴细胞绝对计数、血清LDH和β2-MG水平、ZAP-70表达水平无明显相关性（p〉0．05）,在CD38高表达组中,atm基因缺失的发生率均明显高于CD38低表达组,且差异有统计学意义（p=0．035）。结论：FISH是一种检测CLL患者分子遗传异常快速、准确及敏感的方法,其在CLL患者中的预后预测价值有待进一步的深入研究。     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

目的 探讨典型免疫表型慢性淋巴细胞白血病(CLL)与不典型CLL在临床特征、Binet分期、淋巴细胞绝对计数(ALC)、ZAP-70蛋白表达、CD38表达、IgVH突变和遗传学特性等预后因素上的差异.方法 参照英国CLL临床指南诊断评分系统,77例患者中积分5分的有61例,为典型CLL,积分为4分或3分的有16例,为不典型CLL.采用多参数流式细胞术(FCM)对77例CLL患者的外周血或骨髓标本进行免疫表型检测,包括CD5、CD19、CD23、FMC7、slg(κ和λ)、CD20、CD79b,并检测预后相关因素ZAP-70和CD38的表达水平;采用多重RT-PCR检测IgVH基因突变状态;组合探针荧光原位杂交(FISH)技术检测分子遗传学异常.结果 典型CLL与不典型CLL两组患者在性别、年龄、IgVH基因突变率、ZAP70表达上的差异均无统计学意义(P值分别为0.398、0.189、0.268和0.131);不典型CLL组中ALC≥50×109/L、Binet B+C期和CD38 阳性率≥30%所占比例(分别为43.8%、87.5%、43.8%)明显高于典型CLL组(分别为16.4%、36.1%、16.4%)(P=0.026、P<0.01和P=0.026);典型与不典型CLL组的分子遗传学结果也有显著差异,典型CLL组中单独伴有del(13q14)异常的比例(26.8%)大于不典型组(7.6%),而del(17p13)或del(11q22)异常的比例(12.2%)小于不典型组(46.2%)(P=0.022).结论 典型免疫表型CLL与不典型CLL在Binet分期、ALC、CD38表达和遗传学特性上有显著差异.     

慢性淋巴细胞白血病患者脂蛋白酯酶和血清胸苷激酶水平及其与其他预后因素的相关性研究

目的 探讨慢性淋巴细胞白血病(CLL)患者脂蛋白酯酶(LPL)和血清胸苷激酶(TK)水平及其与其他CLL预后因素的相关性.方法 采用RT-PCR方法 检测58例CLL患者外周血标本中LPL的表达水平;增强化学发光法(ECL)和TK1单克隆抗体检测39例CLL患者外周血血清标本中TK1浓度;多重PCR及序列测定检测IgVH基因突变;流式细胞术检测ZAP-70蛋白及CD38的表达;组合探针荧光原位杂交(FISH)技术检测分子遗传学异常.结果 CLL患者LPL中位表达水平为0.26(0～6.29),而在正常对照中均未检测到LPL表达.LPL表达水平与IgVH基因突变、Binet分期、CD38表达和遗传学异常具有显著相关性,无IgVH突变患者的LPL表达水平明显高于突变患者(P=0.010);Binet A期患者LPL表达水平低于Binet B期和C期患者(P=0.011);CD38高表达组(≥30%)LPL表达水平高于CD38低表达组(<30%)(P=0.001);分子遗传学预后较好组[仅有del(13q14)]LPL表达水平明显低于预后较差组[del(17p13)或del(11q22)](P=0.002).LPL表达水平与患者性别、年龄及ZAP-70蛋白表达水平均无明显相关性(P>0.05).CLL患者血清TK1浓度明显高于正常对照(P<0.05).在外周血淋巴细胞绝对计数(ALC)明显增高组(≥50×109/L)、血清乳酸脱氢酶(LDH)高水平组、无IgVH基因突变组和ZAP-70蛋白高表达组(≥20%)的患者中,血清TK1浓度分别明显高于ALC无明显增高组(<50×109/L)(P=0.018)、血清LDH水平正常组(P=0.018)、具有IgVH基因突变组(P=0.030)和ZAP-70蛋白低表达组(<20%)(P=0.038)的患者.血清TK1浓度与CLL患者性别、年龄、Binet分期、CD38表达水平和遗传学异常无明显相关性(P>0.05).结论 CLL患者LPL和血清TK1表达水平与CLL其他预后因素相关,两者均对IgVH突变情况有一定预示作用,在CLL的预后中具有重要价值.     

59例慢性淋巴细胞白血病分子遗传学异常的FISH检测

目的探讨间期荧光原位杂交(FISH)技术检测慢性淋巴细胞白血病(CLL)常见分子遗传学异常的意义。方法采用CSF12(12p11.1~12q11.1)、D13S25(13q14.3)、RB1(13q14)、ATM(11q22.3)及p53(17p13.1)等5种探针对59例初诊CLL患者的骨髓细胞进行间期FISH检测,分析上述分子遗传学异常与患者的临床Binet分期、Ria分期及相关实验室检查,包括初诊时外周血淋巴细胞绝对值计数、血红蛋白(Hb)水平、血小板计数(PLT),血清乳酸脱氢酶(LDH)、β2-微球蛋白(β2-MG)、CD38和ZAP70水平的相关性。结果 59例CLL初诊患者中,存在1种及1种以上分子遗传学异常的有49例,占83.1%。其中,del(13q)34例(69.4%),12号染色体三体(+12)和p53基因缺失各11例(22.4%),ATM基因缺失5例(10.2%)。各分子遗传学异常与患者临床分期、外周血淋巴细胞计数绝对值、Hb、PLT、LDH、β2-MG、CD38和ZAP70表达水平之间均无明显相关性(P0.05)。≥60岁患者del(13q)异常率(67.6%)显著高于60岁患者(40.9%),女性患者p53基因缺失的异常率(35.2%)显著高于男性患者(11.9%),差异均有统计学意义(P0.05)。结论 FISH技术是检测CLL患者分子遗传学异常快速、准确、敏感的方法,分子遗传学异常与患者初诊时临床表现之间无明显相关性。     

126例慢性淋巴细胞白血病免疫表型分析

目的分析慢性淋巴细胞白血病(CLL)的免疫表型特征。方法应用系列相关单克隆抗体通过三色/四色流式细胞术对126例CLL进行免疫表型分析。结果126例CLL中117例为B系来源(92.9%),9例为T系来源(7.1%)。117例B-CLL均表达CD19,其他B系抗原CD20、CD22、CD23阳性率分别为88.6%、66%、74.2%。无一例表达FMC7和CD10。所有B-CLL中CD5 B-CLL占79.1%,CD5-B-CLL占20.9%。45例B-CLL进行CD38及ZAP-70的检测。共14例CD38 ZAP-70 ,4例CD38 ZAP-70-,3例CD38-ZAP-70 ,24例CD38-ZAP-70-。CD38与ZAP-70的表达具有相关性(P<0.05)。9例T-CLL患者均为成熟T细胞表型,仅表达T系抗原,不表达任何B系抗原。结论了解CLL的免疫表型特点对此类疾病的诊断及治疗后微小残余病灶(MRD)的检测具有重要意义。而免疫表型与预后的关系需作进一步研究。     

分化抗原簇CD38在慢性淋巴细胞白血病T淋巴细胞中的表达研究

目的 探讨慢性淋巴细胞白血病(CLL)中T淋巴细胞CD38(CD38-T)的表达特征.方法 应用多参数流式细胞术检测83例CLL患者肿瘤细胞中CD38表达(CD38-B)、zeta链相关蛋白-70(ZAP-70)以及CD38-T、CD4/CD8比值的表达情况.结果 在所有患者中,CD38+-T占49.4%(41/83),CD38+-B占50.6%(42/83),CD38+-T和CD38-B表达有高度相关性(r=0.553,P<0.01);表达ZAP-70+CD38+-T和ZAP-70-CD38+-T的患者占67.5%(56/83),CD38-T与ZAP-70的表达有相关性(r=0.349,P<0.01).Binet A期患者中CD38+-T占33.3%(14/42),Binet B+C期患者中CD38+T占65.9%(27/41);XD38+-T与临床分期,CD4/CD8比例倒置有相关性(r=0.312、0.453,P<0.05).结论 CD38-T表达变化与患者病情进展及转归密切相关,CD38高表达的患者多伴有免疫功能调节紊乱或失衡,可作为CLL的一项全新独立的监测指标.     

p53和ATM基因缺失在慢性淋巴细胞白血病患者中的预后价值

目的 探讨p53和ATM基因缺失在慢性淋巴细胞白血病患者(CLL)中的预后价值.方法 采用间期荧光原位杂交(FISH)技术和p53、ATM基因的序列特异性DNA探针对80例CLL患者的染色体标本进行p53和ATM基因缺失的检测,分析p53和ATM基因缺失与外周血淋巴细胞绝对计数、Binet分期、血清乳酸脱氢酶(LDH)水平、β2微球蛋白(β2-MG)水平、ZAP-70表达、CD38表达、含氟达拉滨治疗疗效之间的相关性,单因素生存分析采用Kaplan-Meier法绘制生存曲线和Log-rank检验,多因素生存分析采用COX多元回归分析.结果 80例CLL患者中14例(17.5%)伴有p53基因缺失,9例(11.3%)伴有ATM基因缺失,其中3例(3.8%)同时伴有p53和ATM基因缺失.p53和ATM基因缺失与患者性别、年龄、Binet分期、外周血淋巴细胞绝对计数、血清LDH及β2-MG、ZAP-70表达水平无明显相关性;CD38高表达组中,p53和ATM基因缺失的发生率均明显高于CD38低表达组,且差异有统计学意义(P=0.025和P=0.001).41例患者接受含氟达拉滨方案治疗,32例不伴有p53和(或)ATM基因缺失的患者中,12例(37.5%)获得完全缓解(CR),9例伴有p53和(或)ATM基因缺失的患者均未获得CR(P=0.047).单因素生存分析显示,伴有p53基因缺失组的生存期明显较不伴有缺失组短,经Log-rank检验差异有统计学意义(P＜0.01);而伴有与不伴有ATM基因缺失组的生存期差异无统计学意义(P=0.556).COX多元回归分析显示,p53基因缺失(P:0.014)和CD38表达水平(P=0.017)是判断CLL预后的独立因素.结论 伴有p53和(或)ATM基因缺失的患者,接受含氟达拉滨方案治疗疗效差,FISH检测p53和ATM基因缺失在CLL预后判断和指导临床治疗中具有重要价值.     

荧光原位杂交技术检测慢性淋巴细胞白血病分子遗传学异常

目的 :探讨荧光原位杂交技术(fluorescence in situ hybridization, FISH)在慢性淋巴细胞白血病(CLL)诊断和预后评估中的价值。方法 :采用RB1(13q14.1)、D13S25(13q14.3)、p53(17p13.1)、ATM(11q22.3)及CSP12 5组探针对93例初诊CLL患者进行检测,同期进行常规染色体核型(conventional cytogenetics, CC)分析。分析FISH检测出的遗传学异常与患者的临床Binet分期、Rai分期及其他检测指标的相关性。结果 :93例初诊CLL患者分子遗传学异常检出率为79.6%(74/93),其中13q缺失(13q-)阳性率最高,占45.2%;其次为12号染色体三体(+12)占26.9%、p53缺失(17p-)占19.4%、ATM缺失(11p-)占17.2%。同时伴有2种及2种以上染色体异常患者27例(29.0%),其中13q-伴17p-者8例, 13q-伴11q-者5例, 13q-伴+12者4例。CC检测结果相比较, FISH检测结果中患者的阳性率非常显著高于CC检测结果(χ2=32.127, P0.01)。FISH检测结果与Rai分期没有显著相关性(P0.05),伴17p-的CLL患者Binet分期更晚(P=0.012)。各分子遗传学异常与患者年龄、外周血淋巴细胞计数绝对值和CD38表达水平之间均无显著相关性(P0.05)。女性患者13q-的发生率(65.4%)显著高于男性患者(37.3%)(P=0.015);17p-患者IGHV未突变(U-IGHV)的比例显著高于17p-阴性患者(P=0.013);29.0%的患者表达CD38,且与临床分期及U-IGHV呈显著相关(P值分别为0.027及0.006)。结论 :FISH技术能够大大地提高CLL患者分子遗传学异常的检出率,是常规细胞遗传学有力的补充,对CLL患者的临床分期及预后判断均有重要的应用价值。     

间期荧光原位杂交技术检测慢性淋巴细胞白血病ATM基因缺失

目的探讨慢性淋巴细胞白血病(CLL)中ATM基因缺失及其与其他染色体异常及临床分期的相关性。方法运用间期荧光原位杂交技术(I-FISH)和Spectrum OrangeTM标记的位于11q22.3的序列特异性DNA探针ATM对50例初诊CLL患者的染色体标本进行了ATM缺失的检测,同时检测del(13q14)、del(17p13.1)和免疫球蛋白重链基因重排。临床分期按照Binet分期方法。结果50例患者中有6例(12%)ATM缺失;其中4例伴有其它染色体异常。20例Binet A期患者中,3例(15%)存在异常;10例Binet B期患者中,2例(20%)存在异常;13例Binet C期患者中,1例(7.7%)存在异常。ATM缺失在Binet A、B及C期中无统计学差异(P>0.05)。结论I-FISH与常规染色体分析技术相比是一种快速、准确及敏感的方法,对我国CLL患者的预后预测价值有待进一步的深入研究。     

荧光原位杂交技术在慢性淋巴细胞白血病检测中的应用评估

本研究探讨与评估应用间期荧光原位杂交技术(FISH)检测慢性淋巴细胞白血病(CLL)遗传学异常的价值。应用间期FISH技术检测32例初诊CLL患者的del(13q14.3)、del(11q22.3)、del(17p13.1)、del(13q14)和12号染色体三体,同时对免疫表型不典型的10例初诊患者检测IGH/CCND1融合基因。结果表明,在32例病例组中FISH检测出26例(81.3%)基因异常,包括D13S25缺失14例,RB1缺失11例,12号染色体三体9例,P53缺失6例,ATM缺失4例;涉及1种基因异常的12例,其中12号染色体三体7例,D13S25缺失3例,P53缺失1例,ATM缺失1例;涉及2种基因异常的11例,其中D13S25/RB1缺失的7例,另4例均包含P53缺失;涉及3种以上基因异常的病例3例;10例免疫表型表达CD5+CD23-的初诊患者中2例IGH/CCND1(+)。结论:应用间期FISH技术检测CLL基因组的异常,可大大提高异常染色体的检出率,各基因异常有其不同的特点;IGH/CCND1融合基因的检测在CLL诊断中有重要意义。     

ZAP-70、CD38在慢性淋巴细胞白血病中的表达及意义

目的:检测ZAP-70和CD38在慢性B淋巴细胞白血病(B-CLL)中的表达,探讨其与预后的相关性.方法:采用流式细胞仪检测97例慢性B淋巴细胞白血病患者骨髓中白血病细胞ZAP-70、CD38的表达,结合临床分期、化疗疗效进行分析.结果:(1)B-CLL患者中30.9%ZAP-70、CD38同时阳性,49.5%同时阴性.(2)初诊时处于Binet B+C期的:ZAP-70阴性患者为19.0%,ZAP-70阳性患者为75.0%;CD38阴性患者为30.6%,CD38阳性患者为56.8%.ZAP-70、CD38阳性、阴性表达在Binet分期上有显著差异(P<0.00).(3)化疗治疗患者中:ZAP-70阴性者化疗有效(CR+PR)率为90.0%,ZAP-70阳性者化疗有效率为51.3%;CD38阴性者化疗有效率为84.6%,CD38阳性者化疗有效率为48.5%.ZAP-70、CD38阳性、阴性表达在治疗疗效上有显著差异(P<0.01).结论:(1)B-CLL患者ZAP-70表达与CD38表达呈正相关.(2)B-CLL患者ZAP-70、CD38阳性、阴性的表达与Binet分期有关.(3)B-CLL患者ZAP-70、CD38阳性者较阴性者的化疗疗效差.采用流式细胞术检测ZAP-70、CD38的表达可以为B-CLL的临床分期和预后判断提供参考.     

51例慢性淋巴细胞白血病的免疫表型及细胞遗传学特征研究

本研究探讨慢性淋巴细胞白血病（CLL）的免疫表型及细胞遗传学特征,为其诊断及治疗提供依据。采用一组系列相关单克隆抗体和三色流式细胞术对51例CLL患者进行免疫表型分析,并应用R显带技术对这51例患者的细胞遗传学异常进行了研究。结果表明：51例CLL患者中,CD19和CD23阳性率为96.1%,其后依次为CD5（94.1%）、CD20（82.4%）和CD22（78.4%）,CD38阳性率为23.5%。CD5和CD19共阳性者46例（90.2%）。51例患者的染色体异常检出率为35.3%（18/51）,共检出7种主要异常核型,其中＋12有3例,13q-有2例,其它主要异常有＋14、6q-、t（11;14）、t（14;18）和t（2;7）等。各抗原表达与染色体异常无显著性差异（p〉0.05）。结论：典型CLL免疫表型为CD5、CD19和CD23阳性;常规细胞遗传学（CC）检测CLL染色体异常阳性率偏低;免疫表型结合细胞遗传学异常对CLL诊断及预后有重要价值。     

CLLU1在慢性淋巴细胞白血病患者中的表达及其预后意义

目的研究CLLUl在慢性淋巴细胞白血病（CLL）患者中的表达及其与CLL临床分期、细胞遗传学异常、CD38和ZAPHO之间的关系，并评价其预后意义。方法应用半定量RT-PCR技术检测CLLUl在50例CLL患者中的表达水平；应用三色流式细胞术检测CLL患者骨髓或外周血白血病细胞ZAPHO和CD38的表达；应用间期荧光原位杂交（FISH）技术检测CLL患者细胞遗传学异常。结果50例CLL患者中有26例（52％）表达CLLUl，其中7例（26．92％）在BinetA期，19例（73．08％）在BinetB＋C期，二者表达差异有统计学意义（P〈0．01）；24例CD38^＋CLL患者中，17例（70．83％）CLLUl阳性表达，而在26例CD38-CLL患者中，仅有9例（34．62％）CLLUl阳性表达，二者间差异有统计学意义（P〈0．05）；CLLUl在ZAPHO阳性患者中的表达水平高于ZAP-70阴性患者，但两组差异无统计学意义（P〉0．05）；未发现CLLUl与细胞遗传学异常之间的相关性（P〉0．05）。结论CLLUl与临床分期、CD38密切相关，有可能作为CLL患者预后因素之一。     

ZAP-70在24例B细胞慢性淋巴细胞白血病中的表达研究

为了研究慢性淋巴细胞白血病中zeta链相关蛋白-70（zeta-associated protein-70，ZAP-70）的表达及其与其他预后因素的相关性，应用四色流式细胞术检测24例B细胞慢性淋巴细胞白血病（B-CLL）患者骨髓或外周血白血病细胞ZAP-70和CD38的表达。结果显示：①37．5％B-CLL患者表达ZAP-70，其中Binet A期患者有20％（3／15）表达ZAP-70^＋，Binet B＋C期患者有66．7％（6／9）表达ZAP-70^＋，ZAP-70^＋表达在BinetA期和Binet B＋C期之间有显著差异（P〈0．05）；②29．1％B-CLL患者表达CD38，其中Binet A期患者有3例；在此3例中2例同时伴有ZAP-70^＋，CD38^＋在Binet A期和Binet B＋C期之间无显著差异（P〉0．05）；③83．3％（20／24）患者表达ZAP-70^＋CD38^＋或ZAP-70^-CD38^-，ZAP-70和CD38存在相关性（P〈0．05）。结论：在常规实验室可以采用流式细胞术检测ZAP-70，ZAP-70高表达与B-CLL临床分期、染色体及CD38表达等预后相关因素有一定的相关性。     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

应用荧光原位杂交技术检测慢性淋巴细胞白血病的基因组异常

目的 探讨应用组合荧光原位杂交(panel fluorescence in situ hybridization,panel FISH)技术对慢性淋巴细胞白血病(chronic lymphocytic leukaemia,CLL)基因组异常检测的价值.方法 分别应用序列探针D13S25(13q14.3)、RB1、p53、ATM(11q23)和着丝粒探针12号(CSP12)等5种荧光素标记的DNA探针,对17例CLL患者进行FISH检测,并和常规细胞遗传学检测结果进行比较.结果 17例CLL患者中,常规细胞遗传学检测出1例(1/17)有染色体异常,为49,XX,+3,+8,+18;组合FISH检测出10例(10/17)有染色体异常,包括D13S25缺失4例、ATM缺失2例、p53缺失1例、D13S25合并RB1同时缺失2例、多种异常1例.FISH检测的总检出率高于常规细胞遗传学检测.结论 组合FISH技术是检测CLL患者染色体基因组异常的有效手段,与常规细胞遗传学方法相结合则可明显提高CLL染色体异常的检出率.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.