CD38分子表达与成年人急性B淋巴细胞白血病预后的关系

成年人急性B淋巴细胞白血病（B-ALL）的预后研究集中在细胞遗传学并取得进展,发现一些染色体的改变是其独立的预后因素[1,2]。免疫表型分析也是B-ALL预后的重要因素之一,一些抗原表达与某些特定的遗传学改变相关[3]。国外研究证实CD38是慢性B淋巴细胞白血病（B-CLL）独立的预后因素[4]。     

脐血B淋巴细胞的免疫表型及临床意义

目的分析脐血 B淋巴细胞 ( B细胞 )的免疫表型及其临床意义。方法直接免疫荧光标记 ,流式细胞仪检测并对比研究 2 6例正常足月儿脐血和 15例正常成人外周血 B细胞相关表型表达。结果足月儿脐血中成熟 B细胞表型 CD19+ ,CD2 0 + CD19+ ,CD2 2 + CD19+ ,CD2 3 + CD19+ ,CD4 0 + CD19+ 的表达以及不成熟 B细胞表型 CD10 + CD19+ ,CD5 + CD19+ 的表达 ,除 CD2 3 + CD19+ 和 CD4 0 + CD19+ 外 ,均比正常人外周血高 ( P<0 .0 5 ) ,并且无性别之间差异 ( P>0 .0 5 )。结论与成人外周血比较脐血 B淋巴细胞相对不成熟     

CD117在急性白血病中的表达及临床意义

目的探讨CD117在急性白血病中的表达及其临床意义。方法应用CD45/SSC设门,直接荧光标记法,经流式细胞术对73例急性髓系白血病（AML）和47例急性淋巴细胞白血病（ALL）进行CD117的检测。结果对照组、ALL及AML3组CD117的表达阳性率差异有统计学意义（χ2=41.681,P﹤0.01）。AML组CD117的表达阳性率（58.9%）明显高于对照组（0）及ALL组（8.5%）。CD117/CD34的共表达率ALL组明显低于AML组（4.3%vs45.2%,P﹤0.05）。AML组CD117＋患者的CR率为60.5%,CD117-患者为76.7%,两者比较差异无统计学意义（P〉0.05）;而CD117＋/CD34＋患者的CR率为54.5%,明显低于CD117-/CD34-患者的CR率（88.2%,P﹤0.05）。结论 CD117可作为辅助诊断AML的髓系标志抗原,CD34＋/CD117＋可作为进一步排除ALL的指标。CD117＋/CD34＋可能是AML中一类预后不良的特殊亚型,可以作为AML预后判断的指标之一。     

肾衰患者T细胞亚群CD30表达检测及其临床意义

目的 检测慢性肾衰患者(CRF)外周血CD30 T淋巴细胞的表达,以探讨肾衰患者的免疫功能状态.方法 采用流式细胞术检测24例慢性肾衰患者及15例正常对照外周血Th1和Th2的变化.结果 慢性肾衰患者不仅血清中sCD30的表达水平显著增高,外周血循环中CD4 CD30 和CD8 CD30 细胞数亦明显增高,CD4 CD30-和CD8 CD30-细胞均明显下降.但外周血CD4 CD30 和CD8 CD30 细胞的升高与sCD30表达水平的升高并没有明显的相关性,亦未发现CD30 细胞数的增加与急性移植排斥的发生有关.结论 CRF患者CD30 细胞数显著增加,CRF为Th2免疫反应占优势的疾病.CRF患者体内存在着Th2/Te2型细胞的活化和Th1/Th2的失衡.     

系统性红斑狼疮患者外周血淋巴细胞表达BLyS和CD38的变化

目的:研究系统性红斑狼疮(SLE)患者外周血淋巴细胞表达BLyS和CD38的变化。方法:收集22名SLE患者和14名健康人外周血淋巴细胞, 用流式细胞仪检测外周血淋巴细胞表达BLyS和CD38的变化。结果:SLE患者外周血BLyS⁺淋巴细胞、CD19⁺淋巴细胞和CD19⁺CD38⁺淋巴细胞显著增加, BLyS⁺淋巴细胞增加与CD19⁺CD38⁺淋巴细胞增加呈正相关(r=0.434, P<0.05).结论:SLE患者外周血淋巴细胞表达BLyS和CD19⁺B淋巴细胞表达CD38均显著增加, 且二者增加呈正相关。     

CD40在系统性红斑狼疮外周血淋巴细胞的表达

使用密度离心法分离SLE患者和正常人外周血单个核细胞 (PBMC ) ,采用流式细胞术检测B淋巴细胞白细胞分化抗原 4 0 (CD4 0 )的表达水平 ,进行SLE患者 (活动期和缓解期 )和正常人之间的比较 ;并进行B淋巴细胞CD4 0表达水平和血清抗dsDNA抗体水平及狼疮活动指数 (SLEDAI)的相关分析。结果表明 ,活动期SLE患者外周血B淋巴细胞比例 (% )和其表达CD4 0的比例 (% )均明显高于缓解期SLE患者和对照组 ,其表达CD4 0的平均荧光强度 (MFI)在活动期SLE患者最高 ,缓解期SLE患者稍低 ,对照组最低 ;相关分析结果表明 ,活动期SLE患者B淋巴细胞CD4 0的表达比例 (% )和强度 (MFI)均与血清抗dsDNA抗体及SLEDAI呈正相关 ,后两者呈高度相关 ;缓解期SLE患者B淋巴细胞表达CD4 0的强度 (MFI)和SLEDAI呈正相关。CD4 0在活动期SLE患者B淋巴细胞的表达增加 ,其水平与疾病活动度有关。     

CD38在儿童急性淋巴细胞白血病中的表达特点和与预后的关系研究

目的 探讨急性淋巴细胞性白血病(ALL)患儿中CD38的表达特点以及与临床预后的关系,以指导个体化治疗.方法 79例儿童急性淋巴细胞性白血病(B系)初发者,入院后均应用流式细胞仪进行免疫分型,并同时进行微小残留病变(MRD)的筛选,当抗原表达符合CD38低表达时,进一步与其他3色抗体(CD10、CD19和CD34)一起进行共4色的抗体组合检测,作为筛选指标进行进一步的监测.根据CD38表达将ALL患儿分为CD38高表达组和CD38低表达组,对两组进行免疫分型特点、危险分层和生存时间的比较,统计分析使用SPSS16.0软件,P＜0.05为差异有统计学意义.结果 初发急性淋巴细胞性白血病(B系)患儿共79例,CD38低表达组为50/79例(63.3％),CD38高表达组为29/79例(36.7％);79例患儿中仅存在CD38/CD10/CD34/CD19一个筛选指标的4例,包含CD38/CD10/CD34/CD19且≥2个其他筛选指标(如TdT/CD10/CD34/CD19、CD66c/CD10/CD34/CD19和CD45/CD10/CD34/CD19等)的46例,无筛选指标的18例.危险分层中CD38低表达、CD38高表达两组分别为低危21/5例,中危14/15例,高危15/9例.CD38低表达组中early Pre-B 33例,Pre-B 12例,Mature-B 5例,CD38高表达组中early Pre-B 21例,Pre-B 5例,Mature-B 3例;两组中CD38高表达组的高危分层明显大于CD38低表达组(F=6.24,P=0.044),并且生存时间明显小于CD38低表达组,差异有统计学意义(x2=5.22,P=0.022).结论 CD38低表达时可作为急性淋巴细胞性白血病的MRD监测指标,CD38高表达组生存时间缩短,可能为ALL预后不良的独立危险因素.     

慢性淋巴细胞白血病患者骨髓中CD200表达及与预后的关系

目的初步探讨慢性淋巴细胞性白血病(CLL)患者骨髓中CD200的表达及其与免疫表型、染色体核型的关系和与预后相关性。方法采用流式细胞术检测40例初诊CLL患者骨髓中CD200的数量。以CD19+CD200+表达低于50%为CD200low组(18例),大于或等于50%的为CD200high组(22例)。结果 CD200low组(18例)年龄、男性人数、初次白细胞(WBC)数量、初次淋巴细胞(LY)数量、初次LY%、淋巴结发生率显著低于CD200high组(22例)(P0.05);CD200low组初次血红蛋白(HB)量高于CD200high组(P0.05)。CD200low组CD19+、CD19+CD23+、CD19+CD160+抗原表达的平均百分数及Zeta链相关蛋白-70(ZAP-70)阳性率明显低于CD200high组(P0.05)。Binet分期中,CD200low组A期概率多于CD200high组,CD200high组大部分处于C期,差异有统计学意义(P0.05)。Rai分期中,CD200low组患者大部分处于Ⅰ期,CD200high组大部分处于Ⅲ期,差异有统计学意义(P0.05)。结论 CD200对诊断疾病、预后判断、实施个体化治疗、延长患者生存期等具有重要意义。     

干性基因在儿童急性B淋巴细胞白血病中的表达及临床意义

目的 本研究旨在探讨初诊急性B淋巴细胞白血病（B-ALL）患儿骨髓中干性基因的表达及其与临床预后指标的关系.方法 应用Real-time PCR检测32例初诊B-ALL患儿（B-ALL组）和5例骨髓象正常儿童（对照组）的骨髓单个核细胞上干性基因（Oct4、JAG1、Nanog、Sox2、Fgf4等）的表达,比较B-ALL组和对照组的差异,在此基础上结合临床资料（如性别、年龄、外周血白细胞、危险度分层等）进行相关性分析.结果 B-ALL组儿童的相关于性基因Nanog、JAG1、CD133-2、CD44和Runx1的表达水平较对照组明显增高,分别为对照组的2.74±1.00、4.31 ±2.41、10.23±3.21、4.66±1.73和7.44±3.01倍（P＜0.01）,Oct4、Sox2、D LL1、Fgf4和COX-2表达水平较对照组明显降低,分别为对照组的15.00％±0.06、13.00％±0.08、18.12％±0.11、4.73％±0.03和9.59％±0.00（P ＜0.01）;初诊B-ALL组干性基因表达与其性别、年龄、外周血白细胞计数、骨髓幼稚细胞比例、FAB分型、细胞遗传学异常、融合基因表达等均无相关性,其中JAG1基因表达与B-ALL危险度分层有明显相关性（r=0.755,P＜0.01）,其余基因表达与危险度分层无明显相关.结论 与正常儿童的骨髓相比较,初诊B-ALL患儿骨髓中干性基因存在异常表达（上调或下调）,其中JAG1基因表达与B-ALL的临床危险度分层显著相关,提示JAG1基因可能是B-ALL的危险因子,可能对判断患儿预后及指导临床治疗有潜在价值.     

流式细胞术检测CD38、CD138在急性白血病、浆细胞骨髓瘤表型分析中的意义

随着血液恶性肿瘤发病率的增高,运用现代技术准确分析并鉴别恶性肿瘤细胞的来源具有重要意义,它关系到临床治疗方案的选择、放化疗效果的判断、预后的评估。当前流式细胞术作为现代检验技术中的新秀正以其特异性强、客观、准确、快速的优点在血液恶性肿瘤的诊断过程中发挥重要作用。急性白血病、浆细胞骨髓瘤是血液系统常见的肿瘤,应用多种手段对其准确地分型和诊断具有重要意义。我们结合临床需要,在以往常规开展的标记免疫分析项目,包括CD3、CD5、CD7、CD13、CD33、CD10、CD19、CD20、CD22、CD14、CD34、HLA-DR、CMPO的基础上,同时将CD38、CD138抗原检测作为常规开展项目,发现其在急性白血病和浆细胞骨髓瘤的分型诊断中具有很好的补充作用。现将我们工作中的病例进行统计,报告如下。     

Comparison of Bcl-2, CD38 and ZAP-70 Expression in Chronic Lymphocytic Leukemia

Chronic lymphocytic leukemia (CLL) was previously considered a uniform disease characterized by autonomous over-expression of bcl-2. Recently the pathogenic role of bcl-2 has been questioned and attention has turned to prognostic subtypes of CLL differing in CD38 and ZAP-70 expression. However, the relationship between bcl-2 and CD38 or ZAP-70 expression remains uncertain and was investigated using flow cytometric immunophenotyping of 50 CLL specimens. CLL cells were consistently bcl-2 positive but varied in expression level: mean fluorescence intensity (MFI) 45-152. Although there was no significant difference in bcl-2 expression between CD38 or ZAP-70 positive and negative specimens, an inverse correlation was identified between percentage of CD38 positive B-cells and bcl-2 MFI when all (p<0.03, r²=0.10) and peripheral blood (p<0.004, r²=0.27) samples were analyzed. While bcl-2 levels do not appear to be a major discriminator between indolent and more aggressive subtypes of CLL, CD38 and bcl-2 expression appear to be interrelated.     

CD38分子的表达及临床应用前景

CD38分子是单链II型跨膜糖蛋白,广泛表达于造血细胞及非造血细胞系,具有许多复杂而又独特的生物学特性及功能。近年来临床研究发现CD38分子是慢性B淋巴细胞白血病的预测因子,自身免疫反应性糖尿病的诊断指标,并可用于艾滋病及巨细胞病毒的检测及系统性红斑狼疮的病情监测。     

Expression of CD5 and CD38 by human CD5− B cells: Requirement for special stimuli

In this study the mode of expression of CD5 by human tonsillar CD5^? B cells after stimulation with different agents was investigated. Resting B cells were separated into CD5⁺ and CD5^? cells and the two cell fractions exposed to phorbol 12-myristate 13-acetate (PMA). CD5^? B cells expressed CD5 and maximum CD5 expression was achieved after approximately 60 h of culture. Based upon the proportions of cells that express CD5 as well as those of the cells surviving in culture, it was calculated that 15–25 % of the total CD5^? B cells were induced to express CD5. Unlike CD5^? B cells, CD5⁺ B cells proliferated vigorously in response to PMA as assessed by [³H] thymidine incorporation and cell cycle analysis in vitro. However, the expression of CD5 by CD5^? B cells was not related to the selective expansion of some CD5⁺ B cells left over as contaminant cells since this occurred in the absence of cell proliferation. Upon exposure to PMA, CD5^? B cells remained in the G0-G1 phases of the cell cycle and did not express the Ki67 antigen or incorporate [³H] thymidine. Furthermore, mitomycin C treatment of the CD5^? B cells did not prevent CD5 expression. Phenotypic studies disclosed that CD5⁺ B cells but not CD5^? B cells expressed CD39. This finding offered the opportunity to carry out an additional control experiment. Separation of the two populations according to the expression of CD39 confirmed the finding obtained by fractionating the cells into CD5⁺ and CD5^? B cells. The cells induced to express CD5 also expressed CD38 that was not detected on resting CD5^? B cells. In this respect, the CD5^? B cells that converted into CD5⁺ cells (inducible CD5⁺ B cells) resembled the cells from the CD5⁺ B cell fractions that up-regulated CD5 and also expressed CD38 upon exposure to PMA alone. Another example of coordinate expression of these two antigens was the finding that exposure to PMA in the presence of recombinant interleukin-4 (rIL-4) resulted in inhibition of the expression of CD5 and CD38. Although virtually all of the tonsillar CD5^? B cells expressed the CD69 activation marker, no cells other than those co-expressing CD5 and CD38 were induced to express CD5 by PMA alone. Resting CD5^? B cells failed to express CD5 and/or CD38 when cultured with PMA in the presence of EL4 T cells and IL-4-free T cell supernatants. Although this combination of stimuli induced a vigorous cell proliferation, the failure to express CD5 and CD38 was not related to cell cycling, since mitomicyn C-treated CD5^? B cells also failed to express CD5 or CD38 when exposed to PMA in the presence of EL4 cells with or without T cell supernatants. Thus, exposure to T cells alone was sufficient to down-regulate CD5 and CD38 expression. Collectively, the above findings indicate that mature CD5^? B cells can follow distinct pathways of differentiation depending upon the nature of the stimuli encountered, and that CD5 expression may identify a special B cell subset or a particular stage of B cell differentiation.     

慢性乙型肝炎患者外周血淋巴细胞CD8+CD38+的检测意义

目的：通过测定慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的表达及门冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、凝血酶原时间（PT），旨在为治疗慢性乙型肝炎提供有用的参考指标。方法：流式细胞术检测38例慢性乙型肝炎、14例肝硬化患者外周血淋巴细胞CD8^＋CD38^＋细胞的百分率；同时测定AST、TBIL和PT。结果：（1）慢性乙型肝炎组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．01，P〈0．01），肝硬化组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．05，P〈0．05）。肝硬化组CD8^＋CD38^＋、CD8^＋细胞均明显低于慢性乙型肝炎组（P〈0．05，P〈0．05）。（2）慢性乙型肝炎轻、中、重各组之间比较，中度组CD8^＋CD38^＋高于轻度组（P〈0．05），重度组CD8^＋CD38^＋、CD8^＋、CD38^＋均高于轻度组（P〈0．05，P〈0．05，P〈0．05）；重度组CD38^＋高于中度组（P〈0．05）；肝硬化组CD8^＋CD38‘均明显低于轻、中、重各组（P〈0．05，P〈0．01，P〈0．01），肝硬化组CD8^＋低于重度组（P〈0．01）。（3）慢性乙型肝炎患者AST、TBIL、PT不正常组CD8^＋CD38^＋均高于AST、TBIL、PT正常组。结论：慢性乙型肝炎患者CD8^＋CD38^＋细胞明显升高，表明慢性乙型肝炎患者细胞免疫处于异常激活状态，并与肝功能损伤有一定的相关性。慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的测定，对病情分析和诊断有一定的临床参考价值。     

CD38 signaling by agonistic monoclonal antibody prevents apoptosis of human germinal center B cells

The present study demonstrates that an agonistic anti-CD38 monoclonal antibody (mAb) (IB4) is capable of preventing apoptosis of human tonsillar germinal center (GC) B cells as measured by either morphological methods on Giemsa-stained cytospin preparations or flow cytometry on propidium iodidestained cells. Two other anti-CD38 mAb (Leu-17 and OKT10) consistently failed to prevent apoptosis in the same cells, even when tested over a wide range of concentrations. Furthermore, exposure of GC B cells to IB4 mAb up-regulates the bcl-2 proto-oncogene product in a manner similar to that observed with CD40 ligand (CD40L). The ability of IB4 mAb to prevent apoptosis of GC B cells was inferior to that of both anti-CD40 mAb and CD40L. No synergistic or additive effects were observed when IB4 mAb was used together with CD40L. Unlike anti-CD40 mAb or CD40L, IB4 mAb neither induced a proliferation of GC B cells nor increased their proliferative response to anti-CD40, CD40L or recombinant interleukin-4, used alone or in combination. The present results are consistent with the recent findings on either the feature of the CD38 molecules to deliver activation signals and on the mechanisms of selection of B cells that operates in the GC.     

抗人CD38单克隆抗体的制备、鉴定及其功能的初步研究

目的：研制抗人CD38抗原分子的单克隆抗体，进一步研究其生物学功能。方法：采用高表达CD38抗原的Daudi细胞免疫Balb/c小鼠；取其脾细胞与SP2/0细胞融合；用间接免疫荧光法进行杂交瘤筛选；流式细胞术、免疫沉淀法与CD38分子原核表达产物的Western-blot分析鉴定单克隆抗体的特异性。以MTT（四甲偶氨唑盐）法检测单克隆抗体抑制细胞增殖以及介导补体杀伤靶细胞的功能。结果：获得了一株抗人CD38分子的单克隆抗体1C6，流式细胞术显示它具有CD38抗原特异的细胞反应谱，其识别的抗原分子质量为45000u。与CD38分子原核表达产物的Western-blot分析表明，其可特异识别CD38分子胞外结构域。MTT法显示其可介导补体杀伤靶细胞。结论：成功制备了抗人CD38分子的单克隆抗体，并进行了初步的功能研究。     

CD16+56在急性髓系白血病中的表达及其意义

为探讨CD16 56抗原在急性髓系白血病(AML)中的表达及其临床意义,本文用流式细胞仪检测42例AML患者的骨髓细胞,分析其细胞形态学(FAB)分型、免疫表型和临床特点。结果显示:42例AML患者中CD16 56阳性7例(16.6%),1例部分表达(2.4%),34例没有表达(80.9%)。CD16 56阳性的AML患者中,FAB分型、免疫表型和临床表现各有特点,多见于M2、M5,髓外浸润常见,大多数预后不良。     

Signaling through murine CD38 is impaired in antigen receptor-unresponsive B cells

CD38 is a 42-kDa membrane associated enzyme which converts NAD into cyclic ADP-ribose (cADPR), a Ca²⁺-mobilizing second messenger, and ADP-ribose (ADPR). Agonistic antibodies to murine CD38 deliver a potent growth co-stimulus to mature splenic B lymphocytes. In this report we demonstrate a striking relationship between CD38-mediated mitogenesis and the ability of surface IgM to promote B cell proliferation. Tolerized B lymphocytes obtained from a double-transgenic mouse model of B cell tolerance do not proliferate in response to antigen stimulation through the Ig receptor or to agonistic anti-CD38 antibodies. Similarly, B-1 cells isolated from the peritoneal cavity of normal mice, and splenic B cells isolated from newborn mice were also unresponsive to both anti-IgM and anti-CD38 stimulation. All of these CD38-unresponsive B cells expressed normal levels of cell surface CD38 and responded to numerous other stimuli. CD38 immunoprecipitated from these B cell populations was normal in size and effectively hydrolyzed NAD, suggesting that the defect in CD38 signaling likely occurs downstream of CD38 itself. Signaling through CD38 and IgM does not always have identical effects on B cells since anti-CD38 cannot deliver inhibitory growth or differentiation signals to normal B cells or immature B cell lines. Nevertheless, the correlative data with these multiple B cell models of unresponsiveness suggests that the signaling pathway utilized by CD38 and IgM intersect, possibly sharing at least one of the crucial components of the Ig receptor signaling cascade.     

Role of the human CD38 molecule in B cell activation and proliferation

Human CD38 is a surface molecule which has been attributed the function of a signaling channel leading to cellular activation and proliferation, an ectoenzyme with multiple function as well as an inducer of Ca²⁺ mobilization from cytoplasmic stores. The effect mediated by CD38 have been studied in different cell populations: the results obtained in human B cells are apparently contradictory, with CD38 simultaneously leading to apoptosis in early B cells while increasing survival in cells derived from lymph node germinal center. Other effects recently reported concern a different potential in terms of signaling in early B cells and derived cell lines or in more detailed disease models of human leukemia, namely B chronic lymphocytic leukemia cells.
To complete the picture of the effects mediated by CD38 in the B cell compartment, we have studied the signals elicited by ligation of the human molecule in mature B cells from circulating pool and also from spleen of normal individuals. The information obtained completes the picture of CD38 and mature B cells, where we also studied the contribution of relevant cytokines involved in maintenance and differentiation of these normal cells, namely IL-1 α, IL-2, IL-4 and IL-6. Our results indicate that human CD38 plays a key role as a co-receptor in mature B cells from normal individuals.     

慢性乙型肝炎患者外周血淋巴细胞CD44v6的表达及意义

目的：研究慢性乙型肝炎及乙型肝炎后肝硬化患者外周血淋巴细胞（PBL）CD44v6的表达情况及意义。方法：用流式细胞术分别检测50名健康体检者、30例乙型肝炎病毒（HBV）携带者、60例慢性乙型肝炎患者及60例乙型肝炎后肝硬化患者PBLCD44v6的表达水平。结果：慢性乙型肝炎患者PBLCD44v6表达明显高于正常对照组和HBV携带组（P〈0.01）;肝炎后肝硬化组明显高于慢性乙型肝炎组（P〈0.01）;HBV携带组与正常对照组之间的差异也有统计学意义（P〈0.01）。结论：CD44v6在慢性乙型肝炎HBV致肝细胞损伤过程中可能起着重要作用。