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。     

间变性淋巴瘤激酶阳性的大B细胞淋巴瘤病理形态学及分子遗传学研究

目的 探讨间变性淋巴瘤激酶(ALK)阳性的大B细胞淋巴瘤的临床病理及分子遗传学特点.方法 对3例ALK阳性的大B细胞淋巴瘤进行光镜观察,采用免疫组织化学EliVision法及分子遗传学方法检测,并结合文献进行分析和讨论.结果 3例患者均为成年男性,年龄32～42岁,平均年龄36.3岁,病变累及淋巴结.镜下观察:淋巴结结构破坏,可见淋巴窦侵犯,肿瘤细胞大,呈免疫母细胞样/浆母细胞样形态.免疫组织化学示肿瘤细胞CD45、CD138、上皮细胞膜抗原、ALK阳性,且ALK蛋白染色呈胞质内颗粒状阳性,CD3、CD20、CD79a和CD30均阴性.3例间期荧光原位杂交均检测到ALK基因易位.结论 ALK阳性大B细胞淋巴瘤是弥漫性大B细胞淋巴瘤的少见独立亚型,好发于中年男性,病变部位以淋巴结多见,具有特征性形态学、免疫表型和分子遗传学特点.

Abstract：

Objective To study clinicopathologic and genetic features of anaplastic lymphoma kinase (ALK)-positive large B-cell lymphoma (LBCL). Methods Light microscopy, EliVision immunohistocheimical method and fluorescence in-situ hybridization were used to evaluate three ALK + LBCL cases recently diagnosed accompanied with a literature review. Results All three cases were male adult patients ( mean age = 36.3 years) with nodal involvement by lymphoma. Histologic evaluation revealed a diffuse effacement of the nodal architecture by the infiltration of tumor cells. Sinusoidal infiltration was seen.The neoplastic cells were large and exhibited the immunoblastic/plasmablastic morphology. By immunohistochemistry, all the cases showed a cytoplasmic granular staining of ALK. They were positive for CD45, CD138, and epithelial membrane antigen ( EMA), but were negative for CD3, CD20, CD79a and CD30. Fluorescence in situ hybridization (FISH) demonstrated the presence of ALK gene translocation in all of the cases. Conclusions ALK + LBCL represents a distinct variant of diffuse large B-cell lymphoma,usually involving lymph node of middle-aged men. The tumor has a immunoblastic/plasmablastic morphology along with a distinct immunophenotypic profile and ALK gene rearrangement.     

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。     

原发胃肠道弥漫性大B细胞淋巴瘤的免疫分型及其与预后的关系

目的 了解原发胃肠道弥漫性大B细胞淋巴瘤的免疫分型,并比较Choi、Tally和Hans 分型及其与预后的关系.方法 复习90例原发胃肠道弥漫性大B细胞淋巴瘤患者的临床及病理资料并进行随访,应用Kaplan-Meier法、Log-rank检验和Cox比例风险回归模型对临床资料、实验室检测结果 进行生存分析及单因素和多因素预后分析.免疫表型检测采用EnVision和EliVision法,选用的抗体有CD20、CD3ε、CDl0、bcl-6、MUM-1、CD5、bcl-2、GCET1、FOXP1、LMO2、BLIMP1和Ki-67等.结果 (1)年龄为27～83岁,中位年龄58岁,男女比为1.31:1;胃肿瘤58例,占64.4%(58/90);肠肿瘤32例,占35.6%(32/90).(2)肿瘤细胞均表达CD20抗原,均不表达CD3ε和CD5;CD10、bcl-6、MUM-1(30%/80%阈值)的表达率分别为17.8%(16/90)、75.6%(68/90)、52.2%(47/90)/43.3%(39/90),GCET1、FOXP1、LMO2的表达率分别为50.0%(45/90)、45.6%(41/90)、23.3%(21/90),bcl-2、BLIMP1的表达率分别为42.2%(38/90)、8.9%(8/90),Ki-67阳性指数20%～95%,中位数为80%.Hans分型:51.1%为生发中心B细胞型(GCB型),48.9%为非GCB型;Choi分型:55.6%为GCB型,44.4%为活化B细胞(ABC)型;Tally分型:34.4%为GCB型,65.6%为非GCB型.(3)67.8%(61/90)的患者接受化疗,68.9%(62/90)的患者接受手术.患者的2、3和5年总体生存率分别为58.5%、52.8%和49.8%,CHOP方案(环磷酰胺+多柔比星+长春新碱+泼尼松)治疗组的2、3和5年总体生存率分别为68.5%、61.2%和52.9%.结论 Hans和Choi分型各亚型比例差别不大,Tally分型中非GCB型较GCB型比例增高.三种分型的各亚型均存在GCB型优于非GCB/ABC型的趋势.Log-rank检验单因素分析提示乳酸脱氢酶(LDH)水平、国际预后指数(IPI)、化疗、手术、B症状、病变数量、临床分期对预后有影响.Cox比例风险回归模型多因素分析提示Hans分型、Choi分型、化疗、手术、LDH和Lugano分期是独立的预后因素.

Abstract：

Objective To study the immunophenotype and prognostic significance of primary gastrointestinal diffuse large B-cell Iymphoma, with reference to Hans, Choi and Tally algorithms. Methods The clinicopathologic features and follow-up data in 90 cases of primary gastrointestinal diffuse large B-cell lymphoma were analyzed by Kaplan-Meier method, Log-rank test and Cox regression model.Immunohistochemistry was carried out using EliVision and EnVision methods for CD20, CD3ε, CD10,bcl-6, MUM-1, CDS, bcl-2, GCET1, FOXP1, LMO2,BLIMP1 and Ki-67. Results The age of patients studied, 64. 4% (58/90) involved the stomach and 35.6% (32/90) involved the intestine. The immunohistochemical findings were as follows: 100% positivity for CD20, 0% for CD3ε and CD5, 17.8% (16/90) for CD10, 75.6% (68/90) for bcl-6, 52. 2% (47/90) for MUM-1 (cut off was 30%), 43.3%(39/90) for MUM-1 (cut off was 80%), 50.0% (45/90) for GCET1, 45.6% (41/90) for FOXP1,23.3%(21/90) for LMO2, 42.2% (38/90) for bcl-2 and 8.9% (8/90) for BLIMP1. The Ki-67 index ranged from 20% to95% (median =80%). According to Hans algorithm, 51.1% of the cases belonged to germinal center B-cell (GCB) subtype and 48.9% belonged to non-GCB subtype. In contrast, Choi algorithm classified 55.6% cases as GCB subtype and 44. 4% as activated B-cell (ABC) subtype.According to Tally algorithm, 34. 4% were of GCB subtype and 65.6% of non-GCB subtype. Most of the patients (67. 8% ,61/90) received chemotherapy and 68.9% (62/90) underwent surgical resection. The overall 2, 3 and 5-year survival rates were 58. 5%, 52. 8% and 49. 8%, respectively. The overall 2, 3 and 5-year survival rates in the CHOP therapy group were 68.5%, 61.2% and 52. 9%, respectively.Conclusions There is no significant difference in ratio between the GCB and non-GCB/ABC subtypes by Hans and Choi algorithms. The non-GCB subtype seems to be more prevalent according to Tally algorithm.Although there is no significant difference in survival between GCB and non-GCB/ABC subtypes by the 3algorithms, GCB subtype tends to show a better survival. In univariate analysis, LDH level, international prognostic index, chemotherapy, surgical resection, B symptoms, number of involved sites and clinical stage are found to have prognostic significance. In multivariate analysis, Choi algorithm, Tally algorithm,chemotherapy, surgical resection, LDH level and clinical stage are independent prognostic factors.     

弥漫大B细胞淋巴瘤免疫表型分型与预后的关系

目的 探讨弥漫大B细胞淋巴瘤（DLBCL）的免疫表型之生发中心B细胞样（GCB）和非GCB两个亚型的特征及其与DLBCL预后的关系。方法 根据肿瘤细胞免疫组织化学EnVision法标记CD10、bc1-6、MUM-1的表达情况,将133例DLBCL分为GCB和非GCB两个亚型。对以下指标的5年总生存率（OS）及5年无进展生存率（PFS）进行了比较：（1）CD10、bc1-6和MUM-1的阳性和阴性病例;（2）GCB亚型与非GCB亚型;（3）不同国际预后指数（IPI）分组中GCB亚型与非GCB亚型的关系。结果 133例DLBCL中,44例（33．1％）CD10阳性,48例（34．6％）bc1-6阳性,60例（45．1％）MUM-1阳性。CD10阳性DLBCL患者的5年OS及PFS均明显高于CD10阴性患者（P=0．041和0．031）;bc1-6阳性DLBCL患者的PFS明显高于bc1-6阴性患者（P=0．044）,MUM．1阳性DLBCL患者的5年0s及PFS均明显低于MUM-1阴性患者（P=0．031和0．028）。GCB型54例（40．6％）,非GCB型79例（59．4％）。GCB型5年OS及PFS均明显高于非GCB型（P=0．004和0．003）。国际预后指数（IPI）0-1分组及2-5分组中,GCB型5年OS及PFS均明显高于非GCB型（IP10-1分组P=0．019和0．014,2-5分组P=0．006和0．009）,其中IPI2-5分组中的非GCB预后最差。结论 DLBCL亚型及其与IPI联合分析可以作为预测患者预后的有效指标。     

弥漫性大B细胞淋巴瘤的临床病理和免疫组织化学特征

目的 探讨弥漫性大B细胞淋巴瘤临床和病理组织特征以及免疫组织化学特异性抗体在其诊断和鉴别诊断中的价值。方法 收集60例弥漫性大B细胞淋巴瘤,总结其临床资料和病理学特点,用免疫组织化学EnVision^TM两步法标记白细胞共同抗原（LCA）、L26、BLA36、CD30和bcl-6抗体。结果 76．7％（46／60）弥漫性大B细胞淋巴瘤的发病年龄集中在40－70岁,淋巴结内外均可累及,90．0％（54／60）患者临床分期为Ⅱ（24／54）、Ⅲ（21／54）、Ⅳ（9／54）期。组织病理形态：中心母细胞淋巴瘤占88．3％（53／60）,免疫母细胞淋巴瘤占3．3％（2／60）,间变性大细胞淋巴瘤占3．3％（2／60）,富于T细胞的B细胞淋巴瘤占5．0％（3／60）。免疫标记LCA、L26、BLA36表达率为100．0％（60／60）,CD30表达率为3．3％（2／60）,bcl-6表达率为95．0％（57／60）。结论 弥漫性大B细胞淋巴瘤是一组异质性肿瘤,侵袭性大,必需结合其组织病理形态和特异抗体的免疫组织化学检测进行诊断和鉴别诊断。     

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。     

睾丸弥漫性大B细胞淋巴瘤58例临床病理及免疫表型

目的 观察睾丸弥漫性大B细胞淋巴瘤(DLBCL)的临床病理及免疫表型特点,探讨其病理诊断及预后.方法 对58例睾丸DLBCL进行回顾性临床病理研究,包括形态学复习、免疫组织化学(EnVision法)染色、EB病毒编码小RNA(EBER) 1/2原位杂交以及预后相关因素分析.结果 58例睾丸DLBCL患者平均年龄62.1岁,中位年龄65岁.多数病程较短,51例(87.9％)就诊时处于临床Ⅰ～Ⅱ期;48例(82.8％)为单侧睾丸受累.12例(20.7％)伴同侧腹股沟淋巴结肿大,少有其他器官累及.52例(89.7％)病理形态学表现为中心母细胞样细胞在睾丸间质内弥漫性浸润,并浸润曲细精管.睾丸白膜受侵、血管浸润分别见于14例(24.1％)和10例(17.2％).Hans分型以非生发中心B细胞型为主(48/58,82.8％).缺乏EB病毒感染.所有病例行病变睾丸切除,35例(60.3％)接受了术后化疗和(或)放疗.随访率为82.8％ (48/58),其中28例(58.3％)患者死亡;1、3、5年总体生存率分别为55.7％、31.6％和27.6％.年龄＞60岁、B症状、血清乳酸脱氢酶水平升高、高临床分期及未接受术后联合治疗者预后差.结论 睾丸DLBCL确诊时以局部病变为主,低临床分期,有一定的病理形态学特征,多为非生发中心型DLBCL,预后不良;术后联合化疗可延长患者的生存期.     

脾脏滤泡树突状细胞肿瘤伴弥漫性大B细胞淋巴瘤

滤泡树突状细胞（follicular dendritic cell，FDC）是淋巴组织系统中一类非淋巴样辅助细胞，它的主要功能是捕获和提呈抗原和免疫复合物。FDC肿瘤是一种少见的肿瘤，迄今为止，已报道的FDC肿瘤不足70例，脾脏是FDC肿瘤好发部位。作者报道1例女性，50岁，脾脏下极孤立性巨大包块，脾切除后诊断为滤泡树突状细胞肿瘤伴弥漫性大细胞淋巴瘤（DLBCL）。     

弥漫性大B细胞淋巴瘤40例细胞病理学分析

目的 探讨细胞病理学诊断弥漫性大B细胞淋巴瘤的可行性和准确性.方法 选择40例由组织活检证实的弥漫性大B细胞淋巴瘤的细胞学病例,包括浅表淋巴结细针穿刺24例、结外包块细针穿刺6例、胸腔积液5例、腹腔积液1例、脑脊液2例、纤支镜刷片2例.对这些病例的临床特点、细胞形态和免疫细胞化学进行分析.结果 所有病例均查见较多中等偏大的淋巴样肿瘤细胞,其免疫表型为40例肿瘤细胞表达CD45(100%),39例肿瘤细胞表达CD20(97.5%),38例表达CD79α (95%),均表现为弥漫阳性,所有病例均不表达CD45RO和PCK,Ki-67增殖指数(proliferative index,PI)均值为52.1%.其中6例用细胞病理材料所作的免疫细胞化学检测中有6例表达CD45,5例表达CD20,6例表达CD79α,均不表达CD45RO和PCK,Ki-67 PI 50%～70%.40例病例中有7例(17.5%)细胞病理诊断为符合弥漫性大B细胞淋巴瘤,16例(40%)诊断为大细胞淋巴瘤,6例(15%)诊断为可疑淋巴瘤,6例(15%)误诊为低分化癌,5例(12.5%)误诊为炎性病变.结论 细胞病理虽然不是诊断弥漫性大B细胞淋巴瘤的的首选方法,但不失为一种新的尝试,仅观察细胞涂片中细胞的形态难以做出准确诊断,需要结合免疫细胞化学,流式细胞术等辅助方法来提高诊断的准确性和可靠性.     

Anaplastic lymphoma kinase-positive diffuse large B-cell lymphoma with a complex karyotype and cryptic 3' ALK gene insertion to chromosome 4 q22-24

Anaplastic lymphoma kinase (ALK)-positive diffuse large B-cell lymphoma (DLBCL) is a rare tumor that is frequently associated with t(2;17)(p23;q23), a translocation fusing the ALK gene at 2p23 to the clathrin heavy chain gene (CLTC) at 17q23. Here, we report a unique case of ALK-positive DLBCL with plasmablastic morphology and focal cytoplasmic granular ALK stain in an HIV-negative 33-year-old man. By conventional karyotyping, the lymphoma cells were near-tetraploid and included 4 structurally normal copies each of chromosomes 2 and 17. Fluorescence in situ hybridization revealed an apparently normal, intact ALK gene on each of the 4 chromosome 2 homologs plus a cytogenetically cryptic ALK gene insertion into 2 of the 4 chromosome 4 homologs at band 4q22-24. The lymphoma cells expressed CD138, lambda light chain, focal and weak CD30, and exhibited aberrant T-cell antigens, including perforin. This case indicates that ALK-positive DLBCL is more heterogeneous at the cytogenetic/molecular level than previously recognized.     

老年人EB病毒阳性弥漫性大B细胞淋巴瘤的临床病理特征

目的 探讨老年人EB病毒阳性弥漫性大B细胞淋巴瘤(简称老年人EBV阳性DLBCL)的临床病理特征。方法 回顾性分析496例DLBCL,应用EBV原位杂交技术,检测病变组织中EBV感染情况,免疫组织化学检测EBV阳性病例中CD10、CD20、CD30、CD79a、bcl-6、bcl-2、MUM-1、CD5、CD3、T细胞内抗原1(TIA-1)以及Ki-67蛋白表达水平;并结合临床病理资料,分析它们之间的相关性。结果 59例DLBCL的EBER阳性,其中48例为老年人。老年人EBV阳性DLBCL患者中位年龄为73岁,绝大部分( 42/48)年龄＞60岁,男女比为1.4∶1;淋巴结病变11例,结外病变18例,39.6％ (19/48)可见淋巴结病变及结外病变;Ann ArborⅠ～Ⅱ期与Ⅲ～Ⅳ期之比为13/35,33.3％ (16/48)的患者肿瘤累及了2个或以上的结外部位,13.9％ (5/36)的患者体能状态较差,23.3％ (7/30)的患者血清乳酸脱氢酶(LDH)增高;国际预后指数(IPI)3 ～5的18例,IPI 0～2的12例;中位生存期为35个月。镜下观察：淋巴结结构或累及的组织结构完全破坏,在不同程度的炎性反应性背景上,可见中心母细胞、免疫母细胞、H/RS细胞样巨细胞弥漫性浸润或散在分布;老年人EBV阳性DLBCL包括大细胞亚型33例,多形性亚型14例,混合亚型1例。免疫表型上几乎全部表达C D20( 47/48)和（或）CD79a (45/45),绝大部分病例(44/47)可见MUM-1的表达,少量病例表达CDI0或bcl-6;bcl-2及CD30的表达率分别为80.0％ (28/35)及28.9％( 11/38)。大多数(33/39)老年人EBV阳性DLBCL都有很高的增殖指数。与非老年人EBV阳性DLBCL相比,除了年龄偏高及bcl-6的表达较低外,其他临床、病理及免疫表型特征差异均无统计学意义。结论 老年人EBV阳性DLBCL是具有一定独特临床病理特征的DLBCL亚型,但与非老年人EBV阳性DLBCL相类似,在病理诊断上还需结合临床、免疫学等资料去鉴别于其他类型的淋巴瘤。     

Immunohistochemical algorithm alone is not enough for predicting the outcome of patients with diffuse large B-cell lymphoma treated with R-CHOP

Gene expression profiling (GEP), which can divide DLBCL into three groups, is impractical to perform routinely. Although algorithms based on immunohistochemistry (IHC) have been proposed as a surrogate for GEP analysis, the power of them has diminished since rituximab added to the chemotherapy. We assessed the prognostic value of four conventional algorithms and the genes in each and out of algorithm by IHC and fluorescence in situ hybridization in DLBCL patients receiving immunochemotherapy. The results showed that neither single protein within algorithms nor the IHC algorithms themselves had strong prognostic power. Using MYC aberrations (MA) either on the genetic or protein levels, we established a new algorithm called MA that could divide patients into distinct prognostic groups. Patients of MA had much shorter overall survival (OS) and progression-free survival (PFS) than non-MA (2-year OS: 56.9% vs. 98.7%; 2-year PFS: 26.8% vs. 86.9%; P < 0.0001 for both). In conclusions, using additional prognostic markers not associated with cell of origin may accurately predict outcomes of DLBCL. Studies with larger samples should be performed to confirm our algorithm and optimize the prognostic system of DLBCL.     

Recurrent diffuse large B-cell lymphoma presenting initially as hemophagocytic syndrome

Hemophagocytic syndrome (HPS) is an extremely rare and life-threatening abnormality, and the cases secondary to B cell lymphoma are rare. We report a case of relapsed diffuse large B-cell lymphoma initially presenting with hemophagocytic syndrome. The patient developed multiple erythematous macules and progressive thrombocytopenia during the treatment, and died two weeks after admission. The HPS presented as an initial manifestation of the relapsed diffuse B-cell lymphoma and the maculea that appeared during the treatment might be a strong predictor of unfavorable outcome.     

Gene expression patterns in AIDS versus non-AIDS-related diffuse large B-cell lymphoma

Diffuse large B-cell lymphoma (DLBCL) is more prevalent and more often fatal in AIDS patients compared to immune-competent individuals. Potential explanations for these differences include distinct tumorigenic mechanisms and/or altered cellular microenvironments. We previously discovered that the TCL1 (T-cell leukemia-1) proto-oncogene is expressed in a high proportion of AIDS-DLBCL compared to DLBCL cases and that aberrant TCL1 expression causes DLBCL in a new transgenic mouse model. Here, we continue to search for other genes that may contribute to the differential pathogenesis of DLBCL in AIDS. Gene subtraction yielded over 1800 potential AIDS-DLBCL candidates, of which about 50% were unknown and not further considered. The remaining 50% of genes were annotated and, when combined with miniarray screening from multiple patient samples, were reduced to 18 candidate genes for extended analysis. These 18 genes showed distinct patterns of expression in both AIDS-DLBCL and DLBCL samples. However, unlike TCL1, none of these genes was preferentially associated with either AIDS-DLBCL or DLBCL. Our data suggest that the increased incidence and severity of AIDS-DLBCL compared to DLBCL is likely due to crippled immune surveillance rather than to markedly different gene expression profiles.