弥漫性大B细胞淋巴瘤中AID蛋白的表达及意义

目的研究活化诱导胞苷脱氨酶(activation-induced cytidine deaminase,AID)在弥漫性大B细胞淋巴瘤(diffuse large Bcell lymphoma,DLBCL)中的表达,探讨其与DLBCL的临床病理特征、免疫组化亚组及预后的关系。方法采用组织芯片免疫组化法检测101例DLBCL中AID、CD10、BCL-6、MUM1、Ki-67的表达,免疫组化双重染色检测AID与CD20、Ki-67的共定位表达。结果 AID在DLBCL的肿瘤性B细胞中阳性表达率为26.7%。101例DLBCL中,AID阳性表达率在年龄≥60岁患者中明显高于<60岁的患者(P<0.05),AID的表达与患者性别、病变部位、形态学变异型、免疫组化亚组无关(P>0.05)。DLBCL中,AID的表达与B细胞分化标记物CD10、BCL-6和MUM1的表达无相关性(P>0.05),与Ki-67增殖指数(Ki-67>50%)呈正相关(P<0.05)。生存分析表明,AID阳性表达者的5年生存率明显低于AID阴性表达者,两者之间的差异具有统计学意义(P=0.042)。结论 AID的阳性表达与DLBCL的不良预后...     

弥漫性大B细胞淋巴瘤中hENT1的表达及意义

目的 探讨hENT1在生发中心B细胞(germinal center B cell-like,GCB)型与非GCB(non-GCB)型弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及意义.方法 采用免疫组化PV 6000两步法检测CD10、BCL-6、MUM1蛋白在DLBCL的表达并对DLBCL进行亚型分类,同时检测hENT1蛋白的表达,探讨免疫组化染色结果和临床病理参数及预后的关系.结果 (1)hENT1蛋白在DLBCL的GCB及non-GCB亚型中表达差异有显著性(P=0.031,P<0.05).(2)hENT1的表达与患者性别、年龄、部位、LDH高低、Ann Arbor分期、有无B症状的差异均无统计学意义.(3)对76例DLBCL患者进行生存分析,中位随访时间21个月.Log-rank检验GCB/non-GCB组累计生存率差异有统计学意义(P=0.010).结论 DLBCL中non-GCB型患者比例较大,预后差.在治疗过程中,检测hENT1的表达为能否使用核苷类药物提供依据.     

STAT3在艾滋病相关弥漫性大B细胞淋巴瘤中的表达及意义

目的：探讨STAT3在艾滋病相关弥漫性大B细胞淋巴瘤(ADIS/HIV-related diffuse large B-cell lymphoma,HIV-DLBCL)中的表达及意义。方法：收集21例HIV-DLBCL组织标本作为实验组。对照组15例标本,分别为6例HIV阴性的DLBCL(non-HIV-DLBCL)、4例HIV阳性淋巴结反应性增生(HIV-reactive hyperplasia,HIV-RH)、5例HIV阴性淋巴结反应性增生(non-HIV-RH)。采用免疫组化检测各样本中STAT3的表达情况,并结合患者的临床病理因素分析。结果：HIV-DLBCL中STAT3总阳性表达率为76.19%,以胞核表达为主占81.25%;non-HIV-DLBCL中STAT3总阳性表达率为66.67%,以胞核表达为主占50%;两组总阳性率及核阳性率差异无统计学意义(P>0.05)。HIV-RH和non-HIV-RH中,STAT3仅表达于淋巴滤泡细胞;HIV-DLBCL中STAT3表达率与良性病变组具有显著性差异(P<0.05)。STAT3在non-GCB型HIV-DLBCL中的表达明显高于GCB型HIV-DLBCL(P<0.01),与性别、年龄、发病部位及临床分期等因素无关(P>0.05)。结论：STAT3在HIV-DLBCL和non-HIV-DLBCL中均呈高表达,且以核表达为主,提示STAT3过表达与DLBCL的发生有关;STAT3表达与HIV-DLBCL的免疫表型有关,因不同免疫表型DLBCL的预后存在差异,提示STAT3可能是HIV-DLBCL的潜在预后标志物。     

原发性睾丸弥漫性大B细胞淋巴瘤的临床病理及预后

目的观察原发性睾丸弥漫性大B细胞淋巴瘤的临床病理、免疫表型特征及患者存活情况,探讨该肿瘤的病理诊断、鉴别诊断及预后。方法按WHO（2001）淋巴瘤分类标准收集14例原发性睾丸弥漫性大B细胞淋巴瘤,中位年龄62岁,按AnnArbor分期标准,Ⅰ期10例,Ⅱ期3例,Ⅳ期1例。11例有随访资料,其中3例存活,最长存活时间86个月;8例死亡,存活时间5～19个月,中位存活时间为11个月。总结14例的组织病理学、免疫表型特征,并进行存活分析。结果单侧睾丸无痛性肿大是最常见的临床表现。形态学变型全部为中心母细胞性。免疫分型,生发中心样B细胞型（GCB型）1例,非生发中心样B细胞型（non—GCB型）13例。10例p53蛋白表达阳性,肿瘤细胞增殖活性高,6例肿瘤细胞表达bcl-2蛋白。存活分析表明,1、2、5年生存率分别为45．5％、17．0％、17．0％。结论原发性睾丸弥漫性大B细胞淋巴瘤多为外周活化的B细胞起源,预后差,易复发和转移;病理活检加免疫表型检测对肿瘤的诊断和鉴别诊断有重要作用。     

表达间变性淋巴瘤激酶蛋白的弥漫性大B细胞淋巴瘤的临床病理和免疫表型观察

目的探讨表达间变性淋巴瘤激酶（ALK）蛋白的弥漫性大B细胞淋巴瘤（DLBCL）的临床病理特点。方法根据2001年版WHO淋巴造血组织肿瘤分类收集945例DLBCL,以LSAB法作ALK-11染色。对阳性病例再用EnVision法作ALK-11染色,仅EnVision法阳性病例为最终纳入病例。对纳入病例标本用LSAB法加做CD20、CD3、CD30、上皮细胞膜抗原（EMA）、粒酶B、T细胞胞质内抗原（TIA）-1和浆细胞（PC）抗体等免疫表型检测,进行IgH基因重排检测并收集随访资料。结果945例弥漫性大B细胞淋巴瘤中仅5例表达ALK蛋白。4例男性,1例女性,年龄34—72岁,全部原发于淋巴结。临床分期Ⅰ期1例、Ⅱ期2例、Ⅲ期2例。5例随访最长32个月,最短4个月。随访截止时死亡4例,死亡病例最长存活时间32个月。表达ALK蛋白的DLBCL包括中心母细胞性2例、免疫母细胞性1例、间变性1例、浆母细胞性1例;2例中心母细胞性、1例免疫母细胞性和1例间变性均表达CD20。浆母细胞性表达K轻链而不表达CD20。5例均检测到IgH基因重排。ALK蛋白表达：在CD20阳性4例中,1例免疫母细胞性为胞膜和胞质阳性,2例中心母细胞性和1例间变性为胞质颗粒状阳性;1例浆母细胞性为胞核和胞质弥漫阳性。结论ALK蛋白阳性表达DLBLC是一种罕见的,临床过程具侵袭性且预后较差的淋巴瘤,可见于浆母细胞性、中心母细胞性、免疫母细胞性和间变性的大B细胞淋巴瘤。发现1例ALK蛋白表达于胞膜和胞质。     

弥漫性大B细胞淋巴瘤基因表达谱的初步研究

目的探讨弥漫性大B细胞淋巴瘤（DLBCL）不同免疫表型组的基因表达谱状况。方法根据CD10、bcl-6和MUM1的表达状况对156例DLBCL进行分组：CD10^＋和（或）bcl-6^＋、MUM1-（第1组）;CD10^＋和（或）bcl-6^＋、MUM1^＋（第2组）;CD10^-和bcl-6^-、MUM1^＋（第3组）。从各组中各选择3例共（9例）临床分期为Ⅳ期的病例标本,另取3例正常扁桃体组织作为对照,采用Affymetrix U133 plus2．0寡核苷酸芯片研究12例样本的基因表达谱。结果通过unsupervised等级聚类分析,12例样本被分成了4组,分别命名为A、B、C、D组。经与免疫表型分组结果对照显示两种分组结果完全一致：A、B、C组分别对应第1、2、3组,D组对应正常对照组。在DLBCL病例组中（A、B、C组）有81个基因显著表达下调,有86个基因显著表达上调。而其中的一个组（B组）虽然具有混合性生发中心B细胞样（GCB,A组）和活化的外周血B细胞样（ABC,C组）DLBCL的免疫表型,但在聚类分析中发现其基因表达谱与A组和C组均不同,有45个基因表达上调,并且有27个特异性表达基因。结论初步结果显示,DLBCL全基因组表达谱在分子水平上有不同的亚群,且可能通过免疫表型来区分。还提示基因表达谱B组DLBCL可能存在除细胞起源以外的不同异质性因素,而这种因素可能与DLBCL的发病机制相关。     

弥漫性大B细胞淋巴瘤研究进展

弥漫性大B细胞淋巴瘤(diffuse large B—cell lymphoma,DLBCL)是一种大B淋巴细胞弥漫性恶性增生性疾病,瘤细胞核至少2倍于正常淋巴细胞核或大于巨噬细胞核。WHO分类(2001年)将DLBCL视为独立病种,认为进一步区分其形态学变型的重复性差,且不能证实与预后的关系,免疫表型和基因表型对区分这些变型的帮助不大,因此建议病理医师可以使用“弥漫性大B细胞淋巴瘤”这一术语作为最后诊断的名称。     

流式细胞术在弥漫性大B细胞淋巴瘤伴大量T细胞反应病理诊断中的应用

目的探讨流式细胞免疫分型技术在弥漫大B细胞淋巴瘤伴大量T细胞反应病例诊断中的应用价值。方法对3例弥漫大B细胞淋巴瘤伴大量T细胞反应病例病理形态、免疫组织化学染色及流式细胞免疫分型进行回顾性分析。结果例1：在CIM5-SSC（Side Scatter,侧向散射角）点图上可见CD45阳性细胞明显分2群,强阳性细胞由T细胞和B细胞共同组成,但以T细胞为主,其中B细胞为免疫球蛋白轻链非限制性表达。而CD45弱阳性细胞基本都是B细胞,且为免疫球蛋白轻链入限制性表达。例2：在CD45-SSC点图中可见CD45阳性细胞因SSC大小不同而分2群,SSC较小的细胞群为T、B细胞混合存在,SSC较大,相当于单核细胞位置的细胞群主要为B细胞,且为免疫球蛋白轻链K限制性表达;例3：再作CD19／κ／CD20,CD19／λ／CD20三重标记发现部分CD19阳性细胞CD20阴性,该群细胞呈κ限制性表达。结论通过多参数、多组合流式细胞免疫分型技术可以从抗原表达减弱或增强、抗原丢失、是否表达非B系抗原、FSC（前向散射光）和（或）SSC的变化以及是否有表达不成熟B细胞的抗原6个方面将肿瘤性B细胞与反应性细胞区分开。     

ALK阳性的弥漫性大B细胞淋巴瘤1例

患者女性,50岁,因发现左锁骨上淋巴结肿大20天入武警浙江省总队医院.查体:左锁骨上可扪及2枚肿大淋巴结,2 cm×2 cm×2.2 cm至2 cm×2 cm×2.8 cm大小,质硬,活动度差,无压痛,与周围组织粘连.右下颌可扪及1枚肿大淋巴结,1.1 cm×1.0 cm×1.0 cm大小,质中无压痛,活动度可,余浅表及深部淋巴结未触及肿大.     

PLK1和DNAPKcs在弥漫大B细胞淋巴瘤中的表达及临床意义

目的探讨PLK1和DNAPKcs在弥漫大B细胞淋巴瘤(DLBCL)中的临床意义及对DLBCL细胞增殖的影响。方法收集50例DLBCL患者的临床资料,并通过对其病理组织标本进行免疫组化检测PLK1和DNAPKcs的表达水平,分析它们的表达相关性及与DLBCL临床病理特征之间的关系。通过siRNA干扰PLK1和DNAPKcs,采用EdU实验观察DLBCL细胞增殖能力的变化。敲减PLK1的表达,Western blot实验检测DLBCL细胞DNAPKcs的表达变化。结果 PLK1和DNAPKcs表达水平在DLBCL病人和两种细胞系FARAGE和OCI-Ly3中显著高于正常人外周血单核细胞(P0.05),促进DLBCL肿瘤细胞的增殖,与Ann Arbor分期、患者是否出现B症状、IPI评分正相关(P0.05),与DLBCL患者年龄、性别无相关性;敲减PLK1后DNAPKcs的表达水平显著降低。结论 PLK1和DNAPKcs可作为DLBCL诊断和预后判断的重要指标。     

Primary diffuse large B-cell lymphomas of the bone: prognostic relevance of protein expression and clinical factors

Diffuse large B-cell lymphomas can be considered primary bone tumors if they are monostotic or polyostotic, affecting multiple skeletal sites without visceral or lymph node involvement. They are rarely considered as extranodal lymphomas or as bone tumors, respectively. To elucidate the prognostic relevance of clinicopathologic characteristics in such disease, we collected a cohort of primary diffuse large B-cell lymphomas of the bone and retrospectively investigated 33 patients. The cohort encompassed the years 1975 to 2004. Protein expression patterns were identified by immunohistochemistry applied to a tissue microarray. The patients included 23 males (mean age, 37 years) and 10 females (mean age, 54 years). Disease stage was I and II in 30 and IV in 3 patients. Within the mean follow-up of 28 months, 6 patients died. Median overall survival was reached after 78 months. Clinical factors favoring a good prognosis were age younger than 53 and administration of chemotherapy. Of the phenotypic markers analyzed (CD10, CD44s, CD138, Bcl-2, Bcl-6, MUM1, and Ki-67), MUM1 expression in more than 10% of the tumor cells and CD10 expression in less than 55% as well as a nongerminal center signature substantiated adverse outcome in a univariate model. In summary, poor survival in PB-DLBCL was clearly predicted in patients older than 53, who had not received chemotherapy, and who demonstrated MUM1 expression and nongerminal center phenotype.     

Prognostic clinicopathologic factors, including immunologic expression in diffuse large B-cell lymphomas

The aim of this study was to assess the clinical significance and potential prognostic value of the expression of a panel of surface markers, proliferating, suppressor and oncogenic proteins in diffuse large B-cell lymphomas (DLBCL). Biopsies were collected from 158 patients with DLBCL and analyzed immunohistochemically for p53, p21/WAF1, bcl-2, cyclin-D1, bcl-6, mdr, CD5, CD30, epithelial membrane antigen (EMA), Ki-67 and c-myc positive tumor cells. Among these, 76 young and middle-aged patients (20-65 years) were selected to investigate the relationship between protein expression, clinical features, and survival. Survival analysis showed that advanced stage, high lactic dehydrogenase level, and high International Prognostic Index (IPI) were poor prognostic factors associated with a shorter overall survival (OS) and disease-free survival (DFS) times. A high p53 expression and low bcl-6 expression were associated with a shorter DFS time. The histological variant type, cyclin-D1+ CD5+ DLBCL, positive epithelial membrane antigen (EMA+) CD30- DLBCL, high bcl-2 expression, and low Ki-67 proliferation activity tended to be associated with worse survival, but the correlations were not statistically significant. In the multivariate analysis, the most significant factors were age, followed by IPI and last p53. The expression of p21/WAF1, mdr, and c-myc proteins did not influence OS and DFS. The expression of p53 and bcl-6 proteins may be useful prognostic indicators in DLBCL. Cyclin-D1+ CD5+ or EMA+ CD30- DLBCL tended to predict a worse survival and may probably bear a significant prognostic value worthy of consideration. Overall, clinical factors appeared to be more important than biologic parameters in determining the prognosis of diffuse large B-cell lymphomas.     

Immunohistochemical analysis of B-cell lymphoma using tissue microarrays identifies particular phenotypic profiles of B-cell lymphomas

AIMS: To validate the applicability of tissue microarray (TM) in immunohistochemical profiling of B-cell lymphoma and to identify particular phenotypic profiles of B-cell neoplasms. METHODS AND RESULTS: Eighty-two diffuse large B-cell lymphomas (DLBL), 54 follicular lymphomas (FL) and 74 mantle cell lymphomas (MCL) were arrayed. Immunohistochemical stains of TM were compared with immunostains of conventional, formalin-fixed and frozen material sections. Concordant staining results were obtained in more than 88% of cases for CD20, CD3, CD5, CD10, CD23, Bcl-2, IgD, secretory differentiation, p53 and p21 expression. Prognostically relevant hot-spot expression of Ki67 yielded concordant results in 71%. Applying TM for characterization of p27KIP1 expression, both typical and blastoid MCL only rarely showed p27KIP1 expression (9% and 15%), whereas 32% of nodal DLBL were p27KIP1-positive, irrespective of high proliferative activity. Among 22 B-cell lymphomas investigated genetically, a p53 + p21- immunophenotype in >20% of tumour cells correlated with p53 locus deletion. CONCLUSIONS: Lymphoma TM allows for immunohistochemical profiling of human B-cell lymphoma with a comparable accuracy to immunohistochemical studies performed on conventional tissue sections. Nodal DLBLs showed significantly more frequent expression of IgD and p27KIP1 than extranodal DLBL. MCL and DLBL frequently showed aberrant p27KIP1 expression. A p53 + p21- immunophenotype in >20% of tumour cells in B-cell non-Hodgkin's lymphoma correlates with p53 gene deletion.     

MLL2 protein is a prognostic marker for gastrointestinal diffuse large B-cell lymphoma

Mixed linage leukemia gene 2 (MLL2) is identified as a novel mutation gene in diffuse large B cell lymphoma (DLBCL). However, the significance of MLL2 protein expression for the prognosis of DLBCL is unclear. In this study, we detected MLL2 protein expression in primary gastrointestinal diffuse large B cell lymphoma (PGI-DLBCL) samples by using tissue microarray immunohistochemistry, and analyzed the correlation between MLL2 protein expression and tumor proliferation activity. In addition, we investigated clinical significance of MLL2 protein expression for PGI-DLBCL prognosis. We found that there was significant difference in MLL2 protein expression between PGI-DLBCL and reactive hyperplasia of lymph node. High expression of MLL2 protein indicated higher clinical stage. In older patients (>60 years) with PGI-DLBCL, MLL2 protein expression was positively correlated with Ki-67 expression and negatively correlated with patient survival. Our data suggest that MLL2 protein is overexpressed in PGI-DLBCL and appears as a prognostic factor for patients of PGI-DLBCL, especially for those older than 60 years old.     

弥漫大B细胞淋巴瘤中Prdx6的表达及其预后意义

目的 探讨过氧化物酶6(peroxiredoxin-6,Prdx6)在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及其预后意义.方法 回顾性分析286例DLBCL中Prdx6的表达及与临床病理特征和预后之间的关系.结果 Prdx6在DLBCL中的表达高于正常淋巴结...     

Expression of Grb2 distinguishes classical Hodgkin lymphomas from primary mediastinal B-cell lymphomas and other diffuse large B-cell lymphomas

Growth factor receptor-bound protein 2 is an adaptor molecule that mediates B-cell receptor (BCR) signaling pathways, but the expression of growth factor receptor-bound protein 2 in lymphoma tissues has not been reported. We sought to characterize growth factor receptor-bound protein 2 protein expression in reactive tonsillar tissues and lymphoma tissues obtained from diagnostic biopsies of classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, diffuse large B-cell lymphoma, nodular lymphocyte predominant Hodgkin lymphoma, and 20 low-grade B-cell lymphomas. Growth factor receptor-bound protein 2 expression was assessed in tissues by immunohistochemistry and in lymphoma cell lines by immunoblotting. In reactive lymphoid tissues, growth factor receptor-bound protein 2 was expressed in the cytoplasm of B-cells and histiocytes but not T-cells. Strong, cytoplasmic growth factor receptor-bound protein 2 expression was seen in the neoplastic cells of follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, splenic marginal zone lymphoma, primary mediastinal large B-cell lymphoma, diffuse large B-cell lymphoma, and nodular lymphocyte predominant Hodgkin lymphoma. In contrast, only 10% of the classical Hodgkin lymphomas showed growth factor receptor-bound protein 2 expression in the neoplastic cells. Growth factor receptor-bound protein 2 protein expression was detected by Western blotting in all lymphoma cell lines tested with higher levels in primary mediastinal large B-cell lymphoma compared with classical Hodgkin lymphoma cell lines. These findings support a role for growth factor receptor-bound protein 2 in the diagnostically challenging workup of classical Hodgkin lymphoma versus primary mediastinal large B-cell lymphoma and warrant further studies to evaluate the biologic significance of growth factor receptor-bound protein 2 in the pathogenesis of classical Hodgkin lymphoma.     

Common and HIV-related diffuse large B-cell lymphomas differ in their immunoglobulin gene mutation pattern

HIV-infected patients are at high risk of developing diffuse large B-cell lymphomas (DLBCL). It is currently unclear whether these lymphomas represent Epstein-Barr virus (EBV)-driven lymphoproliferations that develop in the setting of immunodeficiency, or whether these tumours are more closely related to the DLBCL seen in the general population. To clarify this issue, 12 HIV-related DLBCL from 11 patients were analysed for the presence of clonally rearranged and somatically mutated immunoglobulin heavy chain (IgH) genes and their association with EBV was determined. Eleven of the 12 tumour samples displayed monoclonal rearrangements of the IgH genes, with or without a moderate number of somatic mutations in the CDRII and in the FWIII regions (average four mutations). One patient presented two successive lesions; whereas the initial tumour showed an oligoclonal IgH rearrangement, the lymphoma at relapse proved to harbour a monoclonal B-cell population. Ten of 12 tumour samples expressed the EBV encoded small RNAs (EBERs), and six of these EBV-positive cases displayed, in addition, an expression of the EBV encoded nuclear antigen 2 (EBNA-2). The results obtained from HIV-related DLBCL are at variance to those described for DLBCL occurring in the general population, since the latter contain significantly more somatic IgH mutations in the CDRII and in the FWIII regions and are only rarely associated with EBV. It is concluded from these findings that HIV-related DLBCL represent a distinct group of B-cell lymphomas, a significant fraction of which most likely originates from EBV-driven lymphoproliferations, and that half of the cases derive from pre-germinal centre B-cells. Copyright © 1999 John Wiley & Sons, Ltd.     

Fluorescent nanoparticle-mediated semiquantitative MYC protein expression analysis in morphologically diffuse large B-cell lymphoma

The current World Health Organization (WHO) classification defines a new disease entity of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements, making fluorescence in situ hybridization (FISH) screening for these genes mandatory. In addition, the prognostic significance of MYC expression was reported, with a cut-off value of 40%. However, interobserver discrepancies arise due to the heterogeneous intensity of MYC expression by immunohistochemistry. Moreover, a cut-off value of positivity for MYC protein in diffuse large B-cell lymphoma (DLBCL) varies among studies at present. Here, we applied a high-sensitivity semiquantitative immunohistochemical technique using fluorescent nanoparticles called phosphor-integrated dots (PID) to evaluate the MYC expression in 50 de novo DLBCL cases, and compared it with the conventional diaminobenzidine (DAB)-developing system. The high MYC expression detected by the PID-mediated system predicted poor overall survival in DLBCL patients. However, we found no prognostic value of MYC protein expression for any cut-off value by the DAB-developing system, even if the intensity was considered. These results indicate that the precise evaluation of MYC protein expression can clarify the prognostic values in DLBCL, irrespective of MYC rearrangement.