CD38 ZAP-70在B慢性淋巴细胞白血病中表达及其临床意义的研究术

目的：检测CD38及zeta链相关蛋白-70（zeta-associated protein-70,ZAP-70）在B慢性淋巴细胞白血病（B—CLL）中的表达,探讨其与临床的相关性。方法：采用流式细胞仪检测51例B慢性淋巴细胞白血病（B—CLL）患者骨髓或外周血白血病细胞CD38和ZAP-70的表达,结合其与患者临床分期、病情进展、化疗疗效进行分析。结果：1）44例患者中10例（22．73％）CD38和ZAP-70同时表达阳性,22例（50％）CD38和ZAP-70同时表达阴性。2）31例CD38^-患者中,初诊时处于Binet B＋C期者9例（29．03％）;11例CD38^＋患者中,初诊时处于Binet B＋C期者7例（63．64％）。CD38阳性、阴性表达在Binet分期上无显著差异（P〉0．05）。27例ZAP-70^-患者中,初诊时处于Binet B＋C期者6例（占22.22％）;16例ZAP-70^＋患者中,初诊时处于Binet B＋C期者10例（62．5％）。ZAP-70阳性、阴性表达在Binet分期上有显著差异（P〈0．05）。3）23例患者予氟达拉滨单药或联合CTX治疗,16例CD38^-表达者中,化疗有效（CR＋PR）者11例（68．75％）;7例CD38^＋表达者中,化疗有效者2例（14.29％）。11例ZAP-70^-表达者中,化疗有效者9例（81．82％）;9例ZAP-70^＋表达者中,化疗有效者1例（11．11％）。CD38和ZAP-70阳性、阴性的表达与化疗疗效有显著差异（P〈0．05）。结论：1）B—CLL患者CD38的表达和ZAP-70的表达有关。2）B—CLL患者ZAP-70阳性、阴性的表达与Binet分期有关系,ZAP-70的阳性表达患者较阴性表达者的病情进展快。3）B—CLL患者CD38和ZAP-70的阳性表达者较阴性表达者的化疗疗效差,预后欠佳。采用流式细胞术检测ZAP-70的表达结合CD38可以作为慢性淋巴白血病的临床分型和预后判断的一个参考指标。     

多发性骨髓瘤免疫标志联合国际分期系统分期对临床治疗及预后判断的意义

 【摘要】　目的　探讨多发性骨髓瘤（MM）瘤细胞的免疫标志特征、CD28及国际分期系统（ISS）分期在临床治疗及预后判断的意义。方法　用四色流式细胞仪检测49例初诊及22例治疗组MM患者瘤细胞膜表面免疫标志的表达情况,并将患者依据ISS分期法、综合风险组分期,对疗效、生存期进行评价分析。结果　随着ISS分期升高,治疗有效率降低[Ⅰ、Ⅱ、Ⅲ期有效率分别为83.3 %（4／6）、38.5 %（5／13）、36.5 %（19／52）]（χ2＝4.235,P＝0.04）,总生存期及无进展生存期缩短,组间比较差异有统计学意义（均P＜0.05）。CD28的异常表达率在初诊及治疗组患者之间差异无统计学意义[27（55.1 %）、14（63.4 %）]（P＞0.05）,CD+28组患者较CD-28 组的生存期缩短,差异有统计学意义（P＝0.040）。综合风险组中,高危组较低危组治疗有效率降低,差异有统计学意义（χ2＝5.620,P＝0.018）,总生存期及无进展生存期缩短,差异有统计学意义（P＜0.01）。结论　MM患者高ISS分期、CD+28表达者、高危组预后较差。     

成年人伴CD2表达B系急性淋巴细胞白血病免疫表型特征分析

 目的　了解成年人伴CD2表达B系急性淋巴细胞白血病（CD+2 B-ALL）的免疫表型特征,为临床诊断、治疗及预后判断提供依据。方法　应用流式细胞术及多种单克隆抗体检测18例成年人CD+2 B-ALL及68例CD-2 B-ALL患者的免疫表型,并对其结果进行分析比较。结果　CD+2 B-ALL的发病年龄明显小于CD-2 B-ALL,18例成年人CD+2 B-ALL的大部分表面标志物与CD-2 B-ALL相似,其中CD10表达水平[（73.78±26.67）%]高于CD-2 B-ALL[（52.84±35.25）%],差异有统计学意义（t＝2.35,P＜0.05）,CD33表达水平[（15.46±27.41）%]则低于CD-2 B-ALL[（31.15±27.72）%],差异有统计学意义（t＝2.16,P＜0.05）;所有B-ALL患者都高表达CD34,阳性表达率分别为72.2 %（13／18）和80.9 %（55／68）,差异无统计学意义（χ2＝0.64,P＞0.05）。CD+2 B-ALL的CD20阳性率明显低于CD-2 B-ALL,差异有统计学意义（χ2＝11.38,P＜0.05）。CD+2 B-ALL伴髓系抗原（CD13或CD33）表达率为44.4 %（8／18）,明显低于CD-2 B-ALL 的72.1 %（49／68）,差异有统计学意义（χ2＝4.86,P＜0.05）。结论　成年人CD+2 B-ALL与CD-2 B-ALL具有相似的免疫表型,主要来源于造血干细胞的恶性转化,CD+2 B-ALL伴髓系抗原（CD13、CD33）及CD20表达明显低于CD-2 B-ALL,提示成年人CD+2 B-ALL可能有较好的预后。     

CD38 ZAP-70在B慢性淋巴细胞白血病中表达及其临床意义的研究

目的:检测CD38及zeta链相关蛋白-70(zeta-associated protein-70,ZAP-70)在B慢性淋巴细胞白血病(B-CLL)中的表达,探讨其与临床的相关性.方法:采用流式细胞仪检测51例B慢性淋巴细胞白血病(B-CLL)患者骨髓或外周血白血病细胞CD38和ZAP-70的表达,结合其与患者临床分期、病情进展、化疗疗效进行分析.结果:1)44例患者中10例(22.73%)CD38和ZAP-70同时表达阳性,22例(50%)CD38和ZAP-70同时表达阴性.2)31例CD38-患者中,初诊时处于Binet B+C期者9例(29.03%);11例CD38+患者中,初诊时处于Binet B+C期者7例(63.64%).CD38阳性、阴性表达在Binet分期上无显著差异(P>0.05).27例ZAP-70-患者中,初诊时处于BinetB+C期者6例(占22.22%);16例ZAP-70+患者中,初诊时处于Binet B+C期者10例(62.5%).ZAP-70阳性、阴性表达在Binet分期上有显著差异(P<0.05).3)23例患者予氟达拉滨单药或联合CTX治疗,16例CD38-表达者中,化疗有效(CR+PR)者11例(68.75%);7例CD38+表达者中,化疗有效者2例(14.29%).11例ZAP-70-表达者中,化疗有效者9例(81.82%);9例ZAP-70+表达者中,化疗有效者1例(11.11%).CD38和ZAP-70阳性、阴性的表达与化疗疗效有显著差异(P<0.05).结论:1)B-CLL患者CD38的表达和ZAP-70的表达有关.2)B-CLL患者ZAP-70阳性、阴性的表达与Binet分期有关系,ZAP-70的阳性表达患者较阴性表达者的病情进展快.3)B-CLL患者CD38和ZAP-70的阳性表达者较阴性表达者的化疗疗效差,预后欠佳.采用流式细胞术检测ZAP-70的表达结合CD38可以作为慢性淋巴白血病的临床分型和预后判断的一个参考指标.     

免疫分型联合核型检查对初诊慢性淋巴细胞白血病的诊断价值

 【摘要】　目的　探讨慢性淋巴细胞白血病（CLL）的免疫表型及核型特征,并分析其在初诊CLL的鉴别诊断价值。方法　回顾性分析70例初诊CLL流式细胞术（FCM）免疫表型及其染色体核型特点,结合临床和实验室结果再分析误诊病例。结果　70例初诊CLL免疫表型均表达CD19,其他成熟B细胞相关抗原CD20、CD22阳性率依次为88.5 %（54／61）及 51.9 %（27／52）,T系相关抗原CD5的阳性率77.1 %（54／70）,CD5 CD19非双阳性表达[CD+5 CD+19（-）]CLL共16例（22.9 %）。34例行CD23检测,阳性率为67.6 %（23／34）。 59例常规染色体核型分析（CC）显示异常核型13例（22.0 %）,最常见为-Y、+12、-20、17号染色体异常,复杂核型5例（8.5 %）,无分裂象2例,正常39例;10例行CLL组合探针荧光原位杂交（FISH）检查,异常检出率为60 %,以GIP D13S25异常检出率最高,为66.6 %。CD+5 CD+19（-）CLL与CD5 CD19双阳性表达[CD+5 CD+19（+）]CLL核型异常发生率差异无统计学意义（P＝0.537）。对部分临床过程不符合CLL惰性病程再分析,70例初诊CLL中6例误诊,其中CD+5 CD+19（+）误诊2例,经荧光原位杂交技术（FISH）t（11;14）及CyclinD1确诊为套细胞淋巴瘤（MCL）;CD+5 CD+19（-）CLL误诊4例,分别为脾边缘区淋巴瘤（SMZL）1例、B淋巴系统增殖性疾病（B-LPD）1例和毛细胞白血病（HCL）2例,误诊率为25.0 %,显著高于CD+5 CD+19（+）CLL（P＝0.030）,而CD23的表达显著低于CD+5 CD+19（+）CLL（P＝0.016）。结论　CD+5 CD+19（-）CLL误诊率高;CLL的典型免疫表型特征为CD5、CD19、CD23的共表达,联合CD20、CD22表达强弱及核型分析有助于CLL与其他B-LPD鉴别; CC联合FISH检查可提高异常染色体的识别能力。免疫表型联合染色体分析能提高误诊CLL的检出率。     

肺癌患者外周血中CD+8 CD-28及CD+4 CDhigh25 CDlow127调节性T细胞的表达及其临床意义

 目的　观察肺癌患者外周血CD+8CD+28、CD+8CD-28及CD+4CDhigh25CDlow127调节性T（Treg）细胞的表达水平,并对CD+8CD-28和CD+4CDhigh25CDlow127 Treg细胞行相关性分析,探讨其在肺癌免疫中的作用及临床意义。方法　采用流式细胞术检测60例初治肺癌患者外周血CD+8CD+28、CD+8CD-28及CD+4CDhigh25CDlow127 Treg细胞的表达率,以60名健康体检者作为对照,分析各指标与肺癌生物学及临床特征之间的关系。结果　肺癌组CD+8CD-28 T细胞［（58.430±15.749）%］和CD+4CDhigh25CDlow127 Treg细胞［（7.365±2.025）%］表达均分别显著高于对照组的（41.057±15.436）%、（6.648±1.669）%,差异有统计学意义（t＝6.102,P＜0.05;t＝2.115,P＜0.05）。肺癌组CD+8CD+28 T细胞表达［（41.570±15.739）%］低于对照组［（58.700±15.298）%］,差异有统计学意义（t＝-6.043,P＜0.05）。上述三个指标的表达与性别、年龄及病理学类型无关（P＞0.05）。随着TNM分期增加,CD+4CDhigh25CDlow127 Treg细胞表达逐渐升高,其中Ⅳ期和ⅢA期之间差异有统计学意义（t＝3.898,P＜0.05）。而CD+8CD-28、CD+8CD+28 T细胞的表达与临床分期无关（P＞0.05）。CD+8CD-28与CD+4CDhigh25CDlow127Treg细胞的表达无线性相关（r＝-0.169,P＞0.05）。结论　CD+8CD-28、CD+4CD25high CD127low Treg细胞在肺癌患者外周血呈高表达,CD+8CD+28T细胞呈低表达,这可能是肺癌患者免疫功能低下的原因之一。CD+8CD-28与CD+4CDhigh25 CDlow127 Treg细胞之间无相关性,但两者联合检测可对了解肺癌患者免疫功能,并为肺癌寻找特异性免疫治疗及预后评价提供新的参考。     

新药诱导后自体造血干细胞移植对以荧光原位杂交分层的多发性骨髓瘤患者的价值

 　目的　评价新药诱导后自体造血干细胞移植（ASCT）对荧光原位杂交（FISH）标危及高危多发性骨髓瘤患者生存的影响。方法　回顾性分析74例多发性骨髓瘤患者,均接受以硼替佐米和（或）沙利度胺为主的诱导化疗,按FISH检测结果及诱导后是否接受ASCT分为标危移植组、标危化疗组、高危移植组及高危化疗组4组,通过生存分析分别评价ASCT对标危及高危患者生存的影响。结果　74例患者经新药诱导后总缓解率为91.9 %（68／74）,接近完全缓解（nCR）+ 完全缓解（CR）率为62.2 %（46／74）。FISH标危的患者与高危的患者诱导后缓解率分别为93.2 %（41／44）与86.7 %（26／30）（P＝0.592）,nCR+CR率分别为56.8 %（25／44）与70.0 %（21／30）（P＝0.251）。FISH标危的患者中,接受移植的患者与接受化疗巩固的患者无进展生存（PFS）及总生存（OS）差异均无统计学意义（P值分别为0.642和0.652）,而FISH高危的患者中,接受移植的患者比接受化疗巩固的患者PFS延长19.7个月（P＝0.028）,OS延长12.5个月（P＝0.542）。结论　在以硼替佐米和（或）沙利度胺为主的方案诱导后,ASCT对 FISH标危多发性骨髓瘤患者的PFS和OS均没有影响,但可使FISH高危患者的PFS延长。     

退化细胞ZAP-70在慢性淋巴细胞白血病的表达及预后影响

  目的  探讨退化细胞、ZAP-70在慢性淋巴细胞白血病（CLL）患者外周血中的表达及其对预后的影响。  方法  利用光学显微镜观察退化细胞在92例CLL患者外周血涂片的检出率; 采用流式细胞术检测外周血淋巴细胞ZAP-70的阳性率。分析二者之间的相关性, 并观察这二项指标与疾病进展的关系。  结果  在92例CLL患者中, Binet A期28例（30.5%）, Binet B期44例（47.8%）, Binet C期20例（21.7%）。外周血涂片退化细胞百分比的中位数为19%（3%~48%）。取≥20%为阳性表达, 则退化细胞阳性56例（60.9%）; ZAP-70阳性32例（34.8%）。双因素直线相关分析提示二者呈负相关。退化细胞阴性患者临床预后差, 较阳性患者的8年无进展生存率（PFS）下降（59.4% vs. 86.0%, P < 0.05）, ZAP-70阳性患者较阴性患者的8年PFS也下降（57.8% vs. 79.5%, P < 0.05）; 退化细胞阴性伴ZAP-70阳性患者PFS更低（27.8%, P < 0.05）。  结论  退化细胞阴性和ZAP-70阳性提示CLL预后不良且二者密切相关。      

骨髓增生异常综合征的免疫表型特征分析

 【摘要】　目的　评价免疫表型在骨髓增生异常综合征（MDS）诊断中的价值。方法　采用流式细胞术对27例MDS患者的骨髓细胞进行免疫表型检测。结果　随着MDS疾病的进展,CD+34细胞比例逐渐升高,分别为：难治性贫血／环形铁粒幼细胞性难治性贫血（RA／RAS）7.43 %,难治性贫血伴原始细胞增多（RAEB）36.81 %,难治性贫血伴原始细胞增多转化型（RAEB-T）56.45 %,3组差异有统计学意义（F＝51.197,P＝0.000）,且各组间差异均有统计学意义（P＜0.05）;髓系抗原CD33、CD13、HLA-DR表达逐渐增高,CD14、CD15抗原表达随着疾病的进展逐渐降低,3组间差异有统计学意义（P＜0.05）;B淋巴细胞表面抗原CD19、CD10的表达随着疾病进展而降低;T淋巴细胞表面抗原CD7表达随着疾病进展而增高,分别为RA／RAS 2.63 %、RAEB 10.79 % 和RAEB-T 11.00 %,3组间差异有统计学意义（F＝10.439,P＝0.001）,其中RA／RAS组与RAEB组、RAEB-T组之间差异有统计学意义（P＝0.000,P＝0.001）。结论　检测MDS患者骨髓细胞的免疫表型有助于MDS的诊断、分型和判断预后,从而为治疗提供依据。     

不同慢性淋巴细胞白血病分期体系对中国患者的适用性

 目的　探讨慢性淋巴细胞白血病（CLL）不同分期体系对中国患者的适用性。方法　回顾性分析2000年至2007年就诊于中国医学科学院血液病医院的218例CLL患者,根据不同的分期系统进行分期,并分析这些分期系统的预后价值。结果　Rai分期中0、Ⅰ、Ⅱ、Ⅲ、Ⅳ期患者的5年总体生存（OS）率分别为（80.5±8.0）%、（79.7±6.7）%、（82.6±7.3）%、（46.1±19.7）%、（48.4±7.3）%;0期患者的OS优于Ⅲ、Ⅳ期患者（P均＜0.05）,Ⅰ期患者的OS优于Ⅳ期患者（P＜0.05）,其余各组间OS差异无统计学意义（P均＞0.05）。修改后的Rai分期中低、中、高危三组患者的5年OS率分别为（80.5±8.0）%、（80.9±5.0）%、（49.6±6.5）%;高危组患者的OS低于中、低危组患者（P均＜0.05）,而低、中危组间OS差异无统计学意义（P＞0.05）。Binet分期中A、B、C期患者的5年OS率分别为（79.9±5.3）%、（71.7±7.7）%、（51.6±6.8）%;A期患者的OS优于B、C期患者（P均＜0.05）,而B、C期间OS差异无统计学意义（P＞0.05）。GIMEMA分期中Ⅰ、Ⅱ、Ⅲ、Ⅳ期患者的5年OS率分别为（85.1±4.5）%、（59.4±8.6）%、（64.3±9.5）%、（48.9±14.0）%;Ⅰ期患者的OS优于Ⅱ、Ⅲ、Ⅳ期患者（P均＜0.05）,其余各组间OS差异无统计学意义（P值均＞0.05）。按Montserrat等和法国协作组的定义可将Binet A期的患者分为"冒烟型"和"活动型"CLL,各种分组方法中的"冒烟型"和"活动型"之间在OS上差异均无统计学意义（P值均＞0.05）。结论　目前国外的CLL各种分期方式在中国患者的预后判断上均有不同程度的局限,需进一步探索纳入了新的预后因素的更适合中国患者的预后分期系统。     

70例B细胞慢性淋巴细胞白血病中ZAP-70、CD38的表达

目的：探讨ZETA相关蛋白（ZAP-70）、CD38在慢性B淋巴细胞白血病（B-CLL）病人骨髓及外周血中的表达及两者相关性,并分析其临床意义。方法：采用流式细胞术检测70例B-CLL病人及12例正常人骨髓或外周血ZAP-70、CD38的表达水平。结果：在70例B-CLL病人中,15例ZAP-70阳性表达,40例CD38阳性表达;对照组ZAP-70、CD38均阴性表达。结论：ZAP-70、CD38在B-CLL中表达与预后相关,阳性提示预后不良。     

Zeta-chain associated protein 70 and CD38 combined predict the time to first treatment in patients with chronic lymphocytic leukemia

BACKGROUND: Zeta-chain associated protein (ZAP)-70 has been proposed as a surrogate marker for immunoglobulin heavy-chain variable region (IgVH) mutation in chronic lymphocytic leukemia (CLL), but it is still not clear whether it is an independent prognostic factor. METHODS: The authors evaluated ZAP-70 expression by flow cytometry in 201 untreated patients and correlated ZAP-70 levels with CD38 expression, genetic abnormalities detected by fluorescence in situ hybridization (FISH), and the time from diagnosis to first treatment. RESULTS: Fifty-seven patients (28%) were positive for ZAP-70 (> or = 20%). Positive ZAP-70 status was associated with advanced disease stage, atypical morphology, CD38-positive status, trisomy 12, del(6q), or no detectable abnormalities; negative ZAP-70 status was correlated with del(13q) as a sole abnormality. The treatment-free interval (TFI) was 17.7 months for ZAP-70-positive patients and 44.6 months for ZAP-70-negative patients (P < 0.001). Multivariate analysis in 117 patients identified advanced stage, CD38 > or = 7%, and the absence of del(13q) as a sole abnormality as independent factors for short TFI. Excluding FISH, ZAP-70 status acquired independent prognostic value along with CD38 status. The authors proposed a risk model that combines ZAP-70 and CD38 to identify patients who are likely to progress. When both markers were positive, the TFI was 12 months; when both were negative, the median TFI was 54 months; a median TFI of 26 months was observed in patients who had discordant results (P < 0.00001). CONCLUSIONS: The current findings suggested that both ZAP-70 and CD38 should be tested prospectively in all patients with early-stage CLL.     

Markers of increased angiogenesis and their correlation with biological parameters identifying high-risk patients in early B-cell chronic lymphocytic leukemia

We wondered whether there was any association between the extent of increased angiogenesis and IgV(H) gene mutational status, expression of CD38 and ZAP-70 in early B-cell chronic lymphocytic leukemia (CLL) patients. Circulating levels of vascular endothelial growth factor (VEGF) correlated positively with ZAP-70-expression (P=0.03), CD38- expression (P=0.03) and mutational status of IgV(H) (P=0.005). The same did not apply when correlations were sought with either bone marrow angiogenesis or serum levels of basic fibroblatic growth factor (FGF-2). Studies to determine how to integrate variables reflecting increased angiogenesis with other prognostic markers such as CD38, ZAP-70, IgV(H) status and cytogenetic abnormalities are needed to optimize risk stratification for individual patients.     

CD38 expression level and pattern of expression remains a reliable and robust marker of progressive disease in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) follows a variable clinical course which is difficult to predict at diagnosis. We assessed somatic mutation (SHM) status, CD38 and ZAP-70 expression in 87 patients (49 male, 38 female) with stage A CLL and known cytogenetic profile to compare their role in predicting disease progression, which was assessed by the treatment free interval (TFI) from diagnosis. Sixty (69%) patients were SHM+, 24 (28%) were CD38+ and ten (12%) were ZAP-70+. The median TFI for: (i) SHM + versus SHM- patients was 124 versus 26 months; hazard ratio (HR) = 3.6 [95% confidence interval (CI) = 1.8 - 7.3; P = 0.001]: (ii) CD38- versus CD38+ patients was 120 versus 34 months; HR = 2.4 (95% CI = 1.4 - 5.3; P = 0.02); and (iii) ZAP70- versus ZAP70+ was 120 versus 16 months; HR = 3.4 (95% CI = 1.4 - 8.7; P = 0.01). SHM status and CD38 retained prognostic significance on multivariate analysis whereas ZAP-70 did not. We conclude that ZAP-70 analysis does not provide additional prognostic information in this group of patients.     

Soluble CD138 serum levels are not associated with other poor prognostic markers in patients with B-chronic lymphocytic leukaemia

The clinical course of CLL is highly variable, and survival from the time of diagnosis of CLL can range from months to decades. Novel biological markers such as IgVH mutation, CD38, and ZAP-70 expression have shown to offer important prognostic informations. Few reports deal with the sCD138 levels and bad prognostic factors in patients with CLL, and contrasting data are reported in literature. In our study, we evaluated the serum level of sCD138 in patients with B-CLL and its relationship with other prognostic markers. There was a significant association between advanced Rai stage and serum sCD138 levels in CLL subjects. Patients with Rai stage III-IV had significantly higher levels of sCD138 with respect to controls (48.85 ± 34 ng/ml vs. 31.1 ± 19.34 ng/ml; P < 0.05). We were unable to demonstrate a significant association between sCD138 serum levels and IgVH gene status, ZAP-70 expression, CD38 expression, beta-2 microglobulin, absolute peripheral blood lymphocytosis, haemoglobin or LDH levels. Our finding that high sCD138 serum levels correlates with advanced stages in patients with B-CLL is consistent with the possibility molecule can identify patients with high tumour burden, but the lack of correlation between sCD138 serum levels and markers such the mutation status of IgVH, ZAP-70, and CD38 suggests that sCD138 levels only reflect the clinical stage of disease than the clinical course or progression.     

Downregulated Dicer expression predicts poor prognosis in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is the most common leukemia in the western world. Alterations in microRNAs (miRNAs) expression have been proposed to play a role in CLL pathogenesis. Dicer and Drosha are the main regulators of miRNA biogenesis, and deregulation of their expression has been indicated as a possible cause of miRNA alterations observed in various cancers. To investigate the role of Dicer and Drosha in CLL, we assessed the expression of Dicer and Drosha and their correlation with other prognostic factors, including Binet stages, immunoglobulin heavy chain variable gene (IGHV) mutation status, TP53 mutation status, ZAP-70 protein and CD38 expression level in 165 CLL patients by using real-time polymerase chain reaction methods. Patients with unmutated IGHV genes had significantly lower expression of Dicer than patients with IGHV mutations. The lower expression level of Dicer was also significantly associated with higher level of CD38 and ZAP-70, and more aggressive Binet stage. We also analyzed Dicer expression in different cytogenetic subgroups. Lower Dicer level was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22.3) in contrast to higher level in good risk cytogenetics (deletion in 13q14 as the sole abnormality). Furthermore, the lower expression of Dicer in CLL shows a strong association with shorter overall survival (OS) (P = 0.0046) as well as with reduced treatment free survival (TFS) (P = 0.0006). By contrast, no differences in the expression of Drosha among these groups of patients were observed. Our data suggest that Dicer expression may play an important role in the progression and prognosis of CLL.     

CD38 as a prognostic factor in Chinese patients with chronic lymphocytic leukaemia

B-cell chronic lymphocytic leukemia (CLL) is the most common type of adult leukemia in the Western countries, however, infrequent in the Eastern. It shows a remarkable heterogeneity, with some patients having an almost normal lifespan, others surviving only several years after diagnosis despite intensive therapy. To explore the prognostic significance of CD38 expression in Chinese patients with CLL, multi-parameter flow cytometry was used to detect the expression of CD38 on CD5(+)CD19(+) cells of 147 patients. CD38 expression and its association with some other prognostic factors such as Binet stage, lymphocyte count in peripheral blood, serum lactate dehydrogenase (LDH), beta2-microglobulin (beta2-MG), ZAP-70 expression and cytogenetic abnormalities were analyzed. The Kaplan-Meier method was used to construct survival curves, and results were compared using the log-rank test. Univariate and multivariate Cox regression analyses were used to assess associations between survival time and potential risk factors. Out of the 147 CLL patients, positive expression of CD38 was found in 45 (30.6%) cases. CD38-positivity identified a subgroup of CLL patients with aggressive disease of Binet stage at the time of the test (P=0.036). Furthermore, the presence of higher serum LDH and beta2-MG levels at diagnosis was strongly correlated with CD38-positive (P=0.016 and 0.025, respectively). Prognostically unfavorable cytogenetic abnormalities, including 17p13 and 11q22 deletions, were significantly more frequent in CD38-positive patients than in CD38-negative ones (P=0.047 and 0.001, respectively). There was no significant difference between CD38-positive and CD38-negative groups in molecular cytogenetic aberrations of del(6q23), del(13q14), 14q32 translocation, or trisomy 12. In addition, in CD38-positive patients, the percentage of leukemic cells expressing ZAP-70 protein was not significantly higher than in CD38-negative ones (P=0.120). CD38 expression was associated with poor outcome. Patients with positive expression of CD38 had significantly shorter overall survival (mean, 81 months) than patients without CD38 expression (mean, 179 months) (P=0.015). Univariate analysis showed that serum levels of LDH and beta2-MG, del(17p13) and CD38 expression were the significant factors in determining overall survival (OS). Del(17p13) and CD38 expression were the variables strongly associated with OS by multivariate Cox regression analysis. It was showed that the patients with high level of CD38 expression had poorer outcome; CD38 was a good predictor of OS in Chinese patients with CLL.     

The prognostic evaluation of CLLU1 expression levels in 50 Chinese patients with chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is characterized by the relentless accumulation of monoclonal B cells with the appearance of small mature lymphocytes and a characteristic CD5 and CD19 co-expression immunophenotype. The incidence of CLL is lower in Asian countries than in Western countries, where CLL is the most common leukemia. To investigate CLLU1 expression in CLL and explore the relationship between CLLU1 expression and alternative prognostic markers, we measured CLLU1 expression levels by semiquantitative RT-PCR in a cohort of 50 Chinese patients with CLL. Analyses of IgVH somatic mutational status, ZAP-70 expression, CD38 expression, and chromosomal aberrations were also performed. The expression of CLLU1 mRNA was determined in 26 of the 50 cases (52%), among which 7 at Binet A (7/21, 33.33%) and 19 at Binet B + C (19/29, 65.52%). The expression levels of CLLU1 were significantly increased in B + C CLL patients at Binet stage compared with those at Binet stage A (P = 0.005). Data for the IgVH somatic mutational status were available for 20 patients with known CLLU1 expression. Five (25%) patients, all expressed CLLU1 mRNA, displayed unmutated IgVH gene usage. While in 15 patients (15/20, 75%) with mutated IgVH gene, only 6 were CLLU1 positive. Patients with unmutated IgVH genes expressed higher levels of CLLU1 than did those with IgVH mutations (P < 0.05). Among 24 CD38(+)-CLL cases, 17 (70.83%) were CLLU1 positive, whereas only 9 (34.62%) positive cases were identified in 26 CD38(-)-CLL patients. Thus, the expression of CLLU1 in CD38(+)-CLL was significantly higher than that in CD38(-)-CLL. However, no significant difference of CLLU1 expression was found between ZAP-70(+) (14/22, 63.64%) and ZAP-70(-) (12/28, 42.86%) patients (P > 0.05). We conclude that CLLU1 expression was significantly associated with clinical stages, IgVH somatic mutational status and CD38 expression, and might be an important prognostic factor in CLL patients.     

Risk Factors for Richter Syndrome in Chronic Lymphocytic Leukemia

Richter syndrome (RS) is defined as the transformation of chronic lymphocytic leukemia (CLL) to a more aggressive B-cell lymphoma, most commonly diffuse large B-cell lymphoma. Approximately 5 – 10 % of CLL patients develop this complication during long-term follow-up. Traditional risk factors for future RS include clinical (advanced Rai stage), biological (ZAP-70, CD38, CD49d) and genetic (del17p, del11q) characteristics at the time of CLL diagnosis. The impact of CLL therapy (purine-nucleoside analogue and/or alkylator-based chemoimmunotherapy and kinase inhibitor therapy) on the risk of RS remains controversial. Both heritable (germline) and acquired (somatic) genetic mutations contribute to risk of RS. Germline polymorphisms in genes related to CD38, LRF4, and BCL-2 have been implicated in the development of RS. Somatic mutations contributing to the development of RS include TP53 disruption, c-myc activation, CDKN2A loss and NOTCH1 mutations. This review summarizes recent advances in our understanding of the biological and genetic factors contributing to RS in CLL patients.     

The significance of soluble HLA-G plasma levels as well as messenger HLA-G for B-cell chronic lymphocytic leukemia (B-CLL)

The immunosuppression accompanies B-cell chronic lymphocytic leukemia (B-CLL) but might be also responsible for disease progression by enabling CLL cells to escape from the immunosurveillance. Some particles involved in the regulation of an immune system might represent prognostic value for B-CLL. Recently we found no correlation between HLA-G on messenger and protein level, suggesting that HLA-G is released in soluble form. To confront this hypothesis we characterized soluble HLA-G (sHLA-G) by the prognostic factors in the first cohort of 34 CLL patients. No correlation was observed between sHLA-G levels in ZAP-70(+) and ZAP-70(−) CLL as well as in CD38(+) CLL and CD38(−) CLL patients. Next, we wondered whether gene expression of HLA-G, which represent the whole HLA-G pool in the cell, posses prognostic value for CLL. In the second cohort of 41 CLL patients we assessed messenger levels of HLA-G by the strongest prognostic factors in CLL including cytogenetics, IgVH mutational status, ZAP-70 as well as CD38. No changes of HLA-G expression levels were found in different CLL groups characterized by IgVH gene mutational status, ZAP-70 as well as CD38. We observed no differences in expression of HLA-G in various cytogenetic groups of CLL including del17p, del13q, del11q, +8q, +3q, del14q and del6q when compared to those with normal karyotype or with 12+. Both, mRNA expression of HLA-G and levels of its soluble form in plasma bring no additional prognostic value for B-CLL patients.