CD10、bcl-6和mum-1在弥漫性大B细胞淋巴瘤中的表达及意义

目的 探讨CD10、bcl-6、mum-1在弥漫性大B细胞淋巴瘤(DLBCL)牛发中心样B细胞(GCB)、非生发中心样B细胞(非GCB)分型中的作用及GCB、非GCB分型的意义.方法 收集54例DLBCL总结其临床资料并进行国际预后指数(IPI)分析;采用免疫组化SP法,观察DLBCL石蜡包埋组织中CD10、bel-6、mum-1的表达并进行GCB、非GCB分型.结果 在54例DLBCL的肿瘤组织中CD10、bel-6、mum-1的阳性率分别为42.6%、46.9%、61.1%;GCB型29例,非GCB型25例.CD10(+)GCB 23例,CD10和mum-1(-). bcl-6(+)GCB 6例;CD10和bcl-6(-)非GCB 20例,CD10(-),bcl-6和mum-1(+)非GCB 5例.通过Kaplan-Meier生存曲线分析及Los Rank检验表明:DLBCL的GCB和非GCB分型与患者的生存率分布筹异显著(P<0.05),GCB型DLBCL患者5年生存率高于非GCB型.54例DLBCL中10例IPI 0～1;14例IPI 2;17例IPI 3;13例IPI 4～5.GCB、非GCB分型与IPI具有很好的对应关系,GCB型以IPI<3为主,非GCB型以IPI>3为主.结论 CD10、bcl-6、mum-1蛋白的联合检测,可以用于DLBCL的GCB、non-GCB分犁及诊断,与IPI结合提示GCB型具有较好的预后.     

B淋巴瘤CD23表达研究

目的 研究Ｂ淋巴瘤ＣＤ２３表达及临床意义。方法 用ＳＰ原位免疫组化方法检测ＣＤ２３表达。结果 ２０例Ｂ淋巴瘤中有１８例ＣＤ２３表达,且与恶性程度及生存时间相关。结论 结果支持Ｂ淋巴ＣＤ２３高表达的假设;ＣＤ２３在Ｂ淋巴瘤发生中起作用和该淋巴因子可能对治疗（药理学调节）有重要意义。     

原发性CD5阳性的弥漫性大B细胞淋巴瘤4例临床病理分析

目的 探讨CD5阳性的弥漫性大B细胞淋巴瘤的临床病理特征、诊断、治疗及预后.方法 回顾性分析4例CD5阳性的弥漫性大B细胞淋巴瘤患者的临床资料、组织病理形态及免疫组化结果.结果 4例患者年龄均＞60岁,乳酸脱氢酶和α-羟丁酸脱氢酶水平均显著升高.光镜下分别在纤维组织中、肝窦内、髓腔中及淋巴结内可见成片或弥漫的体积中～大的异型淋巴样细胞浸润.免疫组化示4例CD20、PAX-5和CD5均弥漫(+),Ki-67阳性率约60％～ 80％,CD3、CD10和cyclinD1均(-);3例bcl-2弥漫(+),1例部分(+).随访0.5～10个月,3例患者6个月内死亡.结论 CD5阳性的弥漫性大B细胞淋巴瘤是一种具有高度侵袭性的弥漫性大B细胞淋巴瘤的亚型,主要累及老年人,预后较差,因此正确掌握临床病理及免疫组化特征对该病的诊断具有重要意义.     

The monoclonality of human B-cell lymphomas

Human tissues involved with lymphoma have been examined in frozen sections for immunoglobulin-bearing cells by a technique involving double-label immunofluorescence with mixed anti-kappa and anti-lambda antibodies. F (ab')2 fragments of purified antibodies were employed to avoid any binding via Fc receptors. B cell lymphomas were shown to be composed of monoclonal populations of Ig bearing cells, whereas normal or reactive lymphoid follicles contained a mosaic of Ig-bearing cells derived from multiple clones. Nodules of lymphoma were often surrounded by normal polyclonal B cell populations. We anticipates that the approach described here will be useful in the diagnosis of lymphoma, differentiating it from reactive lymphoid hyperplasia by the demostration of monoclonality. In addition, it should provide a sensitive and reliable tool for investigating the immunobiology of human lymphoma.     

Effect of interferon-alpha on CD20 antigen expression of B-cell chronic lymphocytic leukemia

Chimeric CD20 monoclonal antibody as alternative therapy in relapsed low-grade non-Hodgkin's lymphoma (NHL) has produced responses in nearly 50% of patients. Augmenting CD20 expression on tumor cells and/or inducing its expression may increase the cell kill and effectiveness of antibody therapy. Peripheral blood lymphocytes from 19 patients with B-cell chronic lymphocytic leukemia (B-CLL) were incubated in vitro in the presence of interferon-alpha (IFN-alpha) (500 U/ml and 1,000 U/ml) for 24 and 72 hours. The effect on CD20 expression was studied by flow cytometry. The differences in the percentage positivity, the mean fluorescence intensity (MFI), and the product of percentage positivity and MFI were used to assess upregulation. There was a significant upregulation of CD20 expression on B cells seen at both concentrations after 24-hour priming (p < 0.01). B-CLL cells cultured for 72 hours in the presence of IFN-alpha also showed upregulation of CD20 expression; however, the degree of upregulation was much lower than that seen at 24 hours. There was no statistically significant increase in CD20 antigen expression on normal lymphocytes following cytokine exposure. These results suggest that IFN-alpha priming may augment the effectiveness of antibody therapy by directly upregulating CD20 antigen expression in addition to its indirect action through effector cells of the host.     

Tumor cell expression of CD59 is associated with resistance to CD20 serotherapy in patients with B-cell malignancies

The anti-CD20 chimeric monoclonal antibody rituximab (Rituxan) is used to treat patients with various B-cell tumors, including patients with plasma cell dyscrasias who have CD20+ disease. Many patients with CD20+ disease have either primary unresponsive disease or progress after initially responding to rituximab; therefore, understanding how tumor cells are, or become, resistant to rituximab is of clinical relevance. In this report, we determined whether tumor cells express antigens that block complement-mediated lysis or antibody-dependent cell-mediated cytotoxicity (ADCC) and thereby contribute to rituximab resistance. We demonstrate that expression of the complement regulator CD59 is associated with resistance to rituximab-mediated complement lysis of multiple myeloma (MM) and non-Hodgkin's lymphoma (NHL) cell lines. Moreover, neutralization of CD59 using a blocking monoclonal antibody reversed resistance to rituximab-mediated complement lysis of CD20++ CD59++ ARH-77 MM cells. In addition, we demonstrate the presence of CD59 and rituximab binding on viable tumor cells from patients with MM and Waldenstrom's macroglobulinemia with progressive disease despite rituximab therapy. Last, we also examined MM and NHL B-cell lines, as well as patient tumor cells, for the expression of other antigens that may have a role in blocking ADCC activity, such as Fas ligand (FasL), MUCI, or TRAIL. FasL, MUC1, and/or TRAIL were coexpressed with complement regulators on many of these cells. These studies therefore show that complement regulators, particularly CD59 and antigens that may block ADCC, are present on various B-cell tumors and associated with rituximab resistance in patients. A prospective, clinical study is assessing the role of these antigens in mediating rituximab resistance.     

BiovaxID, a personalized therapeutic vaccine against B-cell lymphomas

Biovest International Inc (a subsidiary of Accentia BioPharmaceuticals Inc), under license from Stanford University, is developing BiovaxID, a personalized therapeutic vaccine against B-cell lymphomas that, in combination with GM-CSF, exclusively targets cancerous B-cells by raising an immune response to tumor-specific immunoglobulin proteins called idiotypes, for the potential treatment of follicular non-Hodgkin's lymphoma (NHL). Phase I and II clinical trials demonstrated the immunogenicity, safety and therapeutic efficacy of BiovaxID. Phase III clinical trials in NHL were ongoing at the time of publication.     

Ig基因重排检测在B细胞性淋巴瘤诊断中的应用

目的探讨Ig基因重排检测在B细胞性淋巴瘤中的诊断价值。方法选取B细胞性淋巴瘤30例、淋巴组织反应性增生30例,提取DNA,应用BIOMED-2引物系统中的47条引物进行PCR扩增,核酸分子异源双链凝胶电泳分析Ig基因重排结果。结果 30例B细胞性淋巴瘤中检测出Ig克隆性重排,包括IGH(A+B)克隆性重排23例,IGK克隆性重排23例,IGL克隆性重排5例,IGH(A+B)+IGK克隆性重排30例,IGHA+IGK克隆性重排29例;在30例淋巴组织反应性增生中未检测到Ig克隆性重排。结论 Ig基因重排是诊断B细胞性淋巴瘤的有用工具。     

The expanding role of rituximab and radioimmunotherapy in the treatment of B-cell lymphomas

The role of rituximab in the treatment of B-cell lymphomas has rapidly emerged from the relapsed setting to first-line combination regimens across the broad range of histologic subtypes. The role of maintenance rituximab in indolent lymphomas after first-line therapy needs to be defined along with the integration of radioimmunotherapy into the first-line therapeutic regimens. As mechanisms of action/resistance to monoclonal antibody therapy are better understood, approaches to predicting response and optimizing combination therapy to overcoming primary and acquired resistance may be developed.     

Advances in the role of cytogenetic analysis in the molecular diagnosis of B-cell lymphomas

Introduction: Cytogenetic abnormalities represent essential determinants of diagnosis and prognosis in B-cell lymphomas. Their theranostic value is increasingly significant with the development of targeted therapies, in order to adapt the treatment at diagnosis as well as when relapse occurs.

Areas covered: As the significance of these biomarkers is influenced by the technology used to detect them, an overview describing the strength and weakness of conventional and emerging technologies is provided. This review also updates the diverse cytogenetic abnormalities found in B-cell lymphomas, emphasizing their value in treatment decision.

Expert commentary: Cytogenetics remains an essential analysis for the diagnostic work-up of lymphomas. As whole genome sequencing becomes more and more affordable routinely, the next challenge will be to recover all the information conveyed by conventional karyotype, including the analysis of the clonal architecture at the single cell level, in whole genome data.     

系统性红斑狼疮患者外周血单个核细胞CD23,CD25,CD19检测的临床应用

目的 :了解系统性红斑狼疮 ( SL E)疾病中 T、B细胞的变化和两者之间的相互关系。方法 :采用 ABC免疫组化法检测了 3 0例 SL E患者治疗前、后外周血单个核细胞 ( PBMC)分化群 CD2 3、 CD2 5、 CD19的表达 ,并与 2 3名正常健康人作对照。结果 :SL E患者 PBMC CD2 3 、CD2 5 表达分别为 ( 7.9± 1.5 ) % ,( 2 7.2± 6.4 ) % ,均较正常组的 ( 1.6± 1.1) % ,( 4 .4± 1.7) %显著增高 ( P<0 .0 1)。经治疗病情稳定后 CD2 3 ( 2 .0± 1.2 ) % ,与正常组比较无显著性差异 ( P>0 .0 5 ) ;而 CD2 5 虽有下降 ( 10 .2± 6.7) % ,但仍较正常组显著增高 ( P<0 .0 1)。PBMCCD19 表达与正常组比较 ,无显著性差异 ( P>0 .0 5 )。活动性 SL E患者 CD2 3 与 CD2 5 表达呈正相关 ( r=0 .4 0 0 8,P<0 .0 5 )。结论 :在 SL E活动期 T、B细胞均被激活 ,共同参与了 SL E免疫调节。B细胞活化与 T细胞激活有关且主要表现为质的异常 ,而 T细胞异常更难以纠正。     

51例原发性结内弥漫性大B细胞淋巴瘤的预后相关因素分析

目的探讨原发性结内弥漫性大B细胞淋巴瘤(N-DLBCL)的预后相关因素.方法收集51例原发于结内的弥漫性大B细胞淋巴瘤,总结其临床资料和组织病理学特征,用免疫组织化学LSAB法(S-P法)研究其免疫表型,使用的抗体有CD20、CD79α、CD45RO、CD3、Bcl-2、Ki-67、CD30、CD15、κ、λ、Cyclin D1、TdT、GFAP、CK、MPO等.对每例患者进行随访观察,并进行存活分析.结果51例DLBCL被重新分型中心母细胞性变型(CB)40例,B免疫母细胞性变型(B-IB)3例,富于T细胞或组织细胞性变型(T/HCRBCL)1例,B细胞性间变性大细胞性变型(B-ALCL)2例,浆母细胞性变型1例,无法分类4例.Bcl-2阳性表达24例(47.1%).Ki-67指数中位数50.0%,35例(68.6%)≥40.0%.有随访资料的35例存活分析表明,2年和5年生存率分别为48.5%和35.3%,国际预后指数(IPI)≥3比IPI＜3的患者5年生存率低(P＜0.01),有B症状比无B症状的患者5年生存率低(P＜0.05),Ki-67指数≥40.0%比＜40.0%的患者5年生存率低(P＜0.05),BCL-2表达和不表达的患者5年生存率差异无统计学意义(P＞0.05).结论 IPI、B症状、Ki-67指数可以作为DLBCL患者的预后因素,而Bcl-2不是预后因素.     

石蜡包埋B细胞淋巴瘤组织克隆性IgH基因重排的检测

目的：探讨克隆性免疫球蛋白重链第三互补决定簇区（IgHCD43）基因重排在B细胞非霍奇金淋巴瘤（B－NHL）诊断方面的价值。方法：采用半巢式PCR、聚丙烯酰胺凝胶电泳（PAGE）及银染技术,检测38例B－NHL、4例T－NHL及10例慢性扁桃腺炎,经福尔马林固定、石蜡包埋病理组织的克隆性IgHCDR3重排。结果：29例B－NHL及1例免疫组化未能明确细胞来源的NHL克隆性IgHCDR3重排阳性;4例T－NHL及10例慢性扁桃腺炎克隆性IgHCDR3重排阴性。结论：克隆性IgHCDR3重排可作为B－NHL与反应性增生鉴别的特生标志,大部分情况下可作为系分化标志。     

冠心病患者外周血中CD19^＋细胞表达水平的意义

目的观察冠心病患者外周血中CD19^＋的表达水平,并探讨CD19^＋与炎症反应和脂质过氧化反应的关系。方法按美国心脏病学会/美国心脏学会冠心病处理指南的诊断标准并经冠状动脉造影检查确诊的冠心病患者60例,分为稳定型心绞痛（SA）组20例,急性冠状动脉综合征（ACS）组40例;并选择冠状动脉造影正常的20例非冠心病患者作为对照组。应用流式细胞术检测所有研究对象外周血中淋巴细胞表面CD19^＋的表达水平,比较各组CD19^＋的表达情况。同时检测C反应蛋白（CRP）和低密度脂蛋白胆固醇（LDL-C）浓度。并对CD19^＋细胞的表达率与CRP滴度和LDL-C进行Pearson直线相关分析。结果 ACS组患者B淋巴细胞CD19^＋表达率明显高于对照组（10.16±3.63）%vs（4.25±0.96）%（P〈0.01）,亦明显高于SA组（10.16±3.63）%vs（5.39±1.36）%（P〈0.01）;SA组CD19^＋细胞表达率和对照组患者CD19^＋细胞表达率相近（5.39±1.36）%vs（4.25±0.96）%（P〉0.05）。所研究对象外周血中CD19^＋细胞的表达水平与CRP滴度（r=0.661,P〈0.01）和LDL-C（r=0.351,P〈0.01）呈正相关。结论随着冠心病病情加重,冠心病患者外周血中B淋巴细胞CD19^＋的表达水平升高,CD19^＋的表达水平可反映冠心病危险分层,CD19^＋与炎症反应和脂质过氧化反应有关系。     

流式细胞术检测B淋巴细胞表面免疫球蛋白轻链及临床意义

目的 探讨流式细胞术检测外周血B淋巴细胞表面免疫球蛋白(Ig)轻链的实验方法和临床意义.方法 采用流式细胞术,以CD19设门,分析了20例健康对照者和13例淋巴瘤患者B细胞表面Ig kappa与lambda型轻链的表达.结果 对照组kappa型轻链阳性B细胞为(51.4±5.2)%,lambda型轻链阳性B细胞为(39.2±4.3)%,两型轻链B细胞之比值为1.31±0.28;淋巴瘤患者B细胞中kappa型轻链8例(61.5%),lambda型轻链5例(38.5%),两型轻链B细胞的比值或者均小于0.01,或者大于56.78,显著超出健康人比值范围.结论 流式细胞术直接检测B细胞表面kappa轻链与lambda轻链的表达有助于鉴别反应性B淋巴细胞增生和肿瘤性慢性B淋巴细胞增生.