小B细胞恶性淋巴瘤形态学和免疫组织化学研究

目的：探讨各种小B细胞恶性淋巴瘤的形态学、免疫表型特征及其鉴别诊断。方法：对15例小淋巴细胞性淋巴瘤（SLL）、3例淋巴浆细胞性淋巴瘤（LPL）、36例滤泡性淋巴瘤（FL）、25例套细胞淋巴瘤（MCL）、7例淋巴结边缘区B细胞淋巴瘤（MZL）和30例黏膜相关淋巴细胞型结外边缘区B细胞淋巴瘤（MALT－MZL）的石蜡切片进行HE形态学观察和CD5、CD10、CD23和cyclinD1等抗体的免疫组织化学分析。结果：各种小B细胞恶性淋巴瘤在组成细胞和组织结构上各具特征；免疫表型：SLL表达CD5（82％）和CD23（80％），FL表达CD10（87％），MCL表达cyclinD1（84％）和CD5（80％），MZL／MALT－MZL和LPL均不表达CD5、CD10、CD23和cyclinD1。结论：各种小B细胞恶性淋巴瘤均是独立疾病，各具形态学和免疫表型特征，结合HE形态学观察和CD5、CD10、CD23、cyclinD1等免疫组化分析有助于正确诊断和鉴别诊断。     

多发性淋巴瘤性息肉病型套细胞淋巴瘤临床病理分析

目的 探讨大肠多发性淋巴瘤性息肉病(MLP)型套细胞淋巴瘤(MCL)的临床病理与免疫组化特点。方法 采用免疫组化EnVision法确定1例肠道MLP／MCL的免疫表型，抗体包括CD5、CD10、CD19、CD20、CD22、CD79α、bcl-6、bcl-2、CD23、CD43、cyclinD1等。结果 末端回肠、右半结肠、直肠分别见多发性息肉。镜下见肿瘤性淋巴细胞呈弥漫型及结节型生长。瘤细胞表达全B细胞标记，CD5 ，CD10-，cyclinD1 ，CD43 ，CD23-，bcl-6-，bcl-2 。结论 MLP是一种罕见的独特的胃肠道恶性淋巴瘤，几乎均为MCL，具有特殊的免疫表型，需与其他类型B细胞淋巴瘤鉴别。MLP具有侵袭性生物学行为，预后较差，应按中高级别恶性淋巴瘤给予系统性联合化疗。     

脾脏非霍奇金淋巴瘤临床病理学研究

目的 探讨脾脏非霍奇金淋巴瘤(SNHL)的形态学特点、免疫表型及鉴别诊断。方法 对39例SNHL进行形态学观察及免疫组化ABC法分析，使用抗体包括CD3、CD45RO、CD56、CD79α、CD20、CD68、Mac387、CD5、CD10、bcl—2、CD23、CD43、cyclinD1、IgM、IgD等。结果 39例SNHL中有B细胞淋巴瘤(BCL)24例，包括小淋巴细胞性淋巴瘤(SLL)4例，套细胞淋巴瘤(MCL)4例，滤泡性淋巴瘤(FL)5例，边缘区淋巴瘤(MZL)6例，弥漫大B细胞性淋巴瘤(DLBCL)5例，其中2例为大多叶核细胞淋巴瘤。T细胞淋巴瘤(TCL)11例，其中肝脾T细胞淋巴瘤2例，非特异性TCL9例。组织细胞性淋巴瘤4例。结论 脾脏淋巴瘤病理类型多样，掌握其形态学特征，熟悉各类型的免疫表型的异同点对诊断与鉴别诊断有重要意义。     

套细胞淋巴瘤的临床病理特征和细胞周期蛋白D1在诊断中的意义

目的 观察套细胞淋巴瘤的临床病理学特征及细胞周期蛋白D1染色在诊断中的意义。方法 对8例淋巴结套细胞淋巴瘤作临床病理观察及随访,LSAB法做免疫表型分析（CD45RO、CD5、CD20、细胞周期蛋白D1、Ki-67、bcl-2）。结果 患者年龄43-78岁（平均年龄57岁）,男女3：1。组织学特点为：（1）淋巴结结构破坏并被单一的淋巴样细胞所取代,淋巴细胞以套区增生性、结节性、弥漫性三种模式增生。（2）淋巴样细胞核有一定的不规则性,染色质中等致密,核分裂象少见,类似中心细胞。其中有3例转变为高度侵袭性的母细胞样变型。所有的病例都呈cyclinD1与bcl-2阳性、CD20阳性、CD45RO阴性、CD5阳性。结论 套细胞淋巴瘤有其特征的形态改变及免疫表型。根据组织病理学特征及cyclin D1阳性,可与其它类型的小B细胞淋巴瘤相鉴别。套细胞淋巴瘤的母细胞样变型也应当与其它变型区别。     

淋巴结细胞毒性自然杀伤/T细胞淋巴瘤

目的 探讨淋巴结细胞毒性自然杀伤（NK/T）细胞淋巴瘤的临床病理学特征。方法 对5例淋巴结细胞毒性NK/T细胞淋巴瘤作临床病理观察及随访、用ISAB法做免疫表型分析（CD35RO、CD8、CD56、CD30、CD20、TIA-1）及EBER1/2原位杂交检测。结果 淋巴结细胞毒性NK/T细胞淋巴瘤的瘤 理组织学特点为：（1）淋巴结结构明显破坏并被瘤细胞所取代：（2）瘤细胞呈多形性;（3）我数肿瘤细胞表达淋巴细胞分化抗原。5例中CD45RO阳性的有4例,其中3例瘤细胞同时呈CD56阳性;1例为无标记细胞性;所有病例的TIA-1和EBER均为阳性。结论 淋巴结细胞毒性NK/T细胞淋巴瘤有特征性的形态改变和免疫表型。提示肿瘤进展及预后不良。     

富于T细胞/组织细胞的B细胞淋巴瘤的诊断

目的：探讨富于T细胞／组织细胞B细胞淋巴瘤（TCRBCL）的诊断。方法：用S－P石蜡免疫组化法检测22例依据形态学诊断的霍奇金淋巴瘤细胞和背景细胞的免疫表型。结果：4／22例是TCRBCL,3例富于T小淋巴细胞,1例富含组织细胞;瘤细胞3例呈中心母细胞样和免疫母细胞样。1例呈腔隙型细胞样,弥漫散在分布。免疫组化瘤细胞呈CD20（＋）、CD15（－）、CD30（－）、CD21（－）、vimentin(-)。背景细胞CD45RO（＋）／CD68（＋）细胞占绝对优势,为浸润细胞的70％－90％;CD20（＋）细胞散在,CD57（＋）稀少。16例为经典型霍奇金淋巴瘤（CHL）,瘤细胞为CD15（＋）（75％）、CD30（＋）（100％）、vimentin( )(19%)、CD21（－）、CD20（－）及CD45（－）,背景细胞CD45RO（＋）和CD20（＋）数量基本相等,CD57（＋）较少。1例为结节性淋巴细胞为主型霍奇金淋巴瘤（NLPHL）,瘤细胞呈CD20（＋）、CD45（＋）、CD30（－）、CD15（－）,而背景细胞中CD57（＋）较多。结论：石蜡免疫组化在TCRBCL诊断中起重要作用,而且也应用于CHL、NLPHL及TCRBCL间鉴别诊断。     

脾边缘区B细胞淋巴瘤临床病理和免疫组化研究

目的：研究脾边缘区B细胞淋巴瘤（SMZL）临床病理和免疫组化特征,为临床治疗和预后提供依据。方法：组织常规制片,应用ABC免疫组化法标记,光镜观察。结果：6例SMZL以脾肿大为主要临床特征,无全身淋巴结肿大,仅1例外围血和骨髓内查出异形淋巴细胞。病理形态显示结节型4例,弥漫型2例,细胞呈现CCL细胞型3例,MBC型2例,淋巴浆细胞型1例,6例均经免疫组化证实。结论：SMZL与MALT型淋巴瘤／淋巴结边缘区B细胞淋巴瘤组织形态和免疫表型相似,但并不完全相同,SMZL与其它B细胞起源的淋巴瘤临床治疗和预后亦不相同。     

146例套细胞淋巴瘤免疫组织化学结果分析

目的：讨论套细胞淋巴瘤的免疫组织化学特征。方法：回顾性分析146例套细胞淋巴瘤的免疫组化结果,并用FISH方法检测1例Cyclin D1阴性病例是否存在t(11;14)易位。结果：套细胞淋巴瘤免疫组化阳性率为CD20：98.6%(144/146);CD79a：100%(146/146);CD5：88.4%(129/146);CyclinD1：99.3%(145/146);PAX-5：100%(122/122);CD43：84%(79/94);Ki67指数：5%~90%,中位数为20%。少部分病例异常表达CD10、Bcl6、CD23、CD56、CD3、CD45RO。FISH检测1例Cyclin D1阴性病例结果为检测到t(11;14)易位形成的IgH/CCND1融合基因。结论：套细胞淋巴瘤存在较为特征性的免疫组化表达模式,并存在异常表达现象。     

原发于骨骼肌的间变性大细胞T细胞淋巴瘤

目的：探讨骨骼肌原发的间变性大细胞淋巴瘤的临床病理特征和免疫表型。方法：采用常规制片和免疫组化(S-P)法检测1例（14岁）骨骼肌原发的间变性大细胞淋巴瘤。结果：肿瘤细胞CD30、ALK-1、CD45RO和CD45阳性；而CD20、EMA、S-100蛋白、desmin和CD68阴性。结论：本例为间变性淋巴瘤激酶（ALK）阳性的间变性大细胞淋巴瘤。骨骼肌原发的间变性大细胞淋巴瘤非常少见，诊断旱应先排除其它肿瘤和其它部位淋巴瘤累及骨骼肌。     

逆转录聚合酶链反应检测石蜡包埋组织中细胞周期蛋白D1 mRNA对套细胞淋巴瘤的诊断价值

目的 探讨用逆转录聚合酶链反应（RT-PCR）法和竞争性RT-PCR法检测套细胞淋巴瘤（MCL）石蜡包埋组织中细胞周期蛋白D1（cyclin D1）蛋白和mRNA在常规病理工作中的可行性及其诊断和鉴别诊断价值。方法 收集淋巴结内MCL38例、对照组包括结内小B细胞淋巴瘤58例（B小淋巴细胞性淋巴瘤14例,淋巴浆细胞性淋巴瘤3例,滤泡性淋巴34例,淋巴结边缘区B细胞淋巴瘤7例）和淋巴结反应性增生病例20例,用免疫组织化学EnVision法和RT-PCR法、竞争性RT-PCR法检测cyclin D1蛋白及其mRNA的表达,以看家基因PGK作为内对照检测RNA。结果 （1）38例结内MCL中,cyclin D1蛋白阳性率为71．1％（27／38）,对照组均为阴性。（2）116例标本中,可检出内对照PGK基因mRNA表达103例（88．8％）。38例MCL中PGK阳性36例（94．7％）。（3）38例结内MCL中,34例可检出cyclin D1 mRNA表达,去除PGK和cyclin D1 mRNA均阴性的2例,MCL中cyclin D1 mRNA表达的阳性率为94．4％（34／36）。对照组中B小淋巴细胞性淋巴瘤1例检出cyclin D1 mRNA表达,其余病例均未检出cyclin D1 mRNA表达。PCR结果全部经测序证实。（4）用竞争性RT-PCR,38例结内MCL中27例可检出cyclin D1 mRNA高表达,去除2例PGK也为阴性的病例,MCL中cyclin D1 mRNA高表达率为75．0％（27／36）。对照组小B细胞恶性淋巴瘤及淋巴结反应性增生无1例有cyclin D1 mRNA高表达。结论 RT-PCR方法和竞争性RT-PCR方法可在石蜡包埋组织中检测cyclin D1 mRNA的表达,均可用于MCL的诊断。     

The value of immunohistochemistry on paraffin wax embedded tissue sections in the differentiation of small lymphocytic and mantle cell lymphomas.

AIMS: To determine whether immunohistochemistry applied to paraffin wax embedded biopsy tissue can be used to distinguish between B-small lymphocytic lymphoma (B-SLL) and mantle cell lymphoma (MCL). METHODS: Formalin fixed, paraffin wax embedded tissue blocks of 12 cases of B-SLL and 12 cases of MCL were retrieved from the files of the Department of Pathology, Southampton University Hospitals Trust. Following antigen retrieval, where appropriate, sections were stained for CD3, CD5, CD20, CD23, CD43, Cyclin D, PGP9.5, and MIB1 using a streptavidin-biotin complex technique. RESULTS: CD20 stained the neoplastic cells of B-SLL and MCL, and CD3 labelled the reactive T cells in these tumours. In B-SLL, the T cells were generally dispersed among the tumour cells, whereas in MCL they often formed bands around tumour cell nodules. CD5 could be detected on T cells, following antigen retrieval. The level of expression on B cells of B-SLL and MCL was generally too low to allow detection in paraffin wax embedded tissues. CD23 stained B-SLL but not MCL. However, it could be detected in only five of the 12 cases of B-SLL. CD43 could be detected in most cases of B-SLL and MCL. It is not, therefore, of value in distinguishing between these tumours. It will, however, help in the differentiation of B-SLL and MCL from other low grade B cell lymphomas, such as follicle centre cell and marginal zone lymphomas. Cyclin D was expressed in all of the MCL but in none of the B-SLL. PGP9.5 showed reactivity in most cases of MCL and much weaker reactivity in B-SLL. The proliferation indexes of MCL were generally higher than those of B-SLL, as measured by MIB1 labelling. Both tumours, however, showed a wide range of values and considerable overlap. CONCLUSION: Staining for Cyclin D is the most reliable immunohistochemical mean of differentiating between B-SLL an MCL. High levels of PGP9.5, expressed in MCL, may be related to the degradation of Cyclin D by the ubiquitin pathway.     

Expression of transmembrane adaptor protein PAG/Cbp in diffuse large B-cell lymphoma: immunohistochemical study of 73 cases

PAG/Cbp is a transmembrane adaptor protein involved in proximal immune signaling. It is expressed in reactive germinal centers (GC) of secondary lymphatic follicles and related malignant lymphomas. We studied PAG/Cbp expression in GC-like and non-GC-like diffuse large B-cell lymphoma (DLBCL) subtypes. Seventy-three cases of DLBCL identified among 155 malignant lymphomas were classified as GC-like DLBCL (CD10+ or CD10-, bcl-6+, and MUM1-) and non-GC-like DLBCL (CD10-, MUM1+ or CD10-, bcl-6+, MUM1+). PAG/Cbp was detected by monoclonal antibody MEM-255 following routine immunohistochemical procedures. Thirty-five of 40 GC-like DLBCLs (88%) and 20 of 33 non-GC-like DLBCL cases (61%) expressed PAG/Cbp. Four of 12 bcl-6-negative non-GC-like DLBCL cases (33%) were PAG/Cbp positive, and only 4 of 20 bcl-6-positive non-GC-like DLBCL cases (25%) were PAG/CBP negative. All 37 FL and all 5 Burkitt's lymphomas (BL) expressed PAG/Cbp, whereas all 6 mantle cell lymphomas (MCL) and 4 of 5 chronic lymphocytic leukemias (CLL/SLL) were PAG/Cbp negative. PAG/Cbp is a reliable GC marker. Its expression correlates with GC-like DLBC phenotype in a significant majority of cases. It is typically absent in MCL and SLL/CLL.     

Large cell variants of CD5+, CD23- B-cell lymphoma/leukemia

CONTEXT: Mantle cell lymphoma (MCL), and its leukemic phase, constitute a well-studied hematologic malignancy with known overall survival, prognostic indicators, morphologic findings at diagnosis and in bone marrow, and known incidence of the bcl-1 immunoglobulin gene rearrangement. Large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype (CD5+, CD23-), including but not limited to blastic MCL, prolymphocytoid MCL, blastic mantle cell leukemia, and prolymphocytic mantle cell leukemia, are not as well characterized. Although blastic MCL is known to be associated with a shorter overall survival than conventional MCL, the large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype have not been described as fully as conventional MCL. OBJECTIVE: The purpose of the present study was to describe the large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype. DESIGN: Nineteen cases of large cell variants of CD5+, CD23- B-cell lymphoma/leukemia are reviewed and described in regard to morphology, bone marrow morphological findings, Cyclin D1 immunostaining, and bcl-1 analysis. Clinical data were not available owing to the varied clinical sources of the specimens. SETTING: Tertiary-care academic institution. RESULTS: Lymph node involvement in blastic CD5+, CD23- B-cell lymphoma was diffuse (100%) with a nodular component (33%) or focal mantle zone pattern (10%). Bone marrow involvement in blastic CD5+, CD23- B-cell lymphoma was seen in only 27% of cases and was composed predominantly of small, slightly irregular lymphocytes. Cyclin D1 was demonstrated in 60% of the 15 cases analyzed and more sensitive in B5-fixed tissue. Bcl-1 (performed in 5 cases) was not detected in the 4 cases of blastic CD5+, CD23- B-cell lymphoma analyzed and was detected in the case of the prolymphocytoid MCL. Cyclin D1 was demonstrated in all 4 bcl-1 negative cases and was negative in the bcl-1 positive prolymphocytoid MCL. CONCLUSION: Careful analysis of clinical data, morphology, immunophenotype, Cyclin D1 expression, and molecular analysis are required to differentiate the unusual large cell variants of MCL from other processes.     

Immunohistochemical detection of follicular dendritic cells in the diagnosis of small B-cell lymphomas

Our study included 46 cases of B-chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL), 25 mantle cell lymphomas (MCL), and 40 follicular lymphomas (FL). The lymphomas were diagnosed according to current morphological, immunohistochemical, and in some cases molecular-genetic criteria. The patterns of follicular dendritie cells (FDCs) were studied in paraffin sections using the anti CD21, anti CD23, and CNA.42 antibodies. The differences in staining patterns of FDCs among the studied lymphomas were statistically significant. The absence or small number of FDC clusters are typical of B-CLL/SLL, while FDC meshwork limited to peripheral parts of neoplastic follicles is characteristic for FL. In contrast, MCL is characterized by irregular FDC meshwork or by FDC clusters present in the centres of residual follicles. Thus, we have demonstrated that the patterns of FDCs is an independent diagnostic feature for distinguishing between B-CLL, FL, and MCL.     

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.     

Lymphomas of the bone: a pathological and clinical study of 54 cases

We examined 28 cases of primary bone lymphomas (PBL; stage IE) and 26 cases of systemic lymphomas involving the bone (SBL; stage IIE to IV). Two histologic types were prevalent: Diffuse large B-cell lymphomas (DLBCL; 26 PBL and 21 SBL) and CD30+ anaplastic large cell lymphomas (ALCL; 1 PBL and 4 SBL). A mature B phenotype (CD45+, CD20+, CD79a+, CDw75+/-, CD10-/+) was established in the DLBCL group. Bcl-2 immunoreactivity was demonstrated in 13/37 cases (35%), and bcl-6 immunostaining was observed in 22/32 cases (69%). ALCL showed null/T phenotype (CD3-/+; CD43+/-; CD30+), with ALK-1 expression in 3/3 cases. With use of a FR3A primer, a monoclonal pattern was demonstrated by PCR analysis in 22/41 lymphomas (54%). Bcl-2 translocation was identified in 2/41 cases (5%). This study details the clinical and pathological characteristics of bone lymphomas. Our immunohistochemical and molecular data suggest that most of them are "de novo" DLBCL and support their follicle center origin.     

CD23 expression in mantle cell lymphoma: clinicopathologic features of 18 cases

The distinction between small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) and mantle cell lymphoma (MCL) has important clinical implications. Typically, SLL/CLL is CD23+, whereas MCL is CD23-. However, CD23 is expressed in a subset of MCLs, and the clinicopathologic features of patients with these neoplasms are not well described. We report 18 CD23+ MCLs, detected by flow cytometry in all cases (dim intensity, 16; bright intensity, 2), 5 (28%), also positive by immunohistochemical analysis. There were 13 men and 5 women (median age, 56 years), 5 of whom died (median survival, 46 months). Seventeen (94%) had bone marrow involvement. Lymphadenopathy (14 cases [78%]), splenomegaly (11 cases [61%]), and leukemic involvement (10 cases [56%]) were common. Five cases (28%) had blastoid morphologic features. The frequency of CD23 expression by MCL is method-dependent, being typically dim and most commonly detected by flow cytometry. In this small study group, bone marrow and leukemic involvement were relatively common.