CD5-positive mucosa-associated lymphoid tissue (MALT) lymphoma of ocular adnexal origin: usefulness of fluorescence in situ hybridization for distinction between mantle cell lymphoma and MALT lymphoma

Extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue type (MALT lymphoma) usually lacks CD5 expression. Herein is described two cases of CD5-positive MALT lymphoma of ocular adnexal origin. The differential diagnosis between CD5-positive MALT lymphoma and mantle cell lymphoma (MCL), notably cyclin D1-negative MCL, was difficult because both cases consisted histologically of small to medium-sized cells with diffuse or vaguely nodular growth pattern, and the neoplastic cells were positive for CD5 and negative for cyclin D1. Somatic mutation analysis of the immunoglobulin heavy chain variable region (VH) gene in case 1 found a relatively higher mutation frequency (5.0%), which was not definitive to rule out MCL. Interphase fluorescence in situ hybridization (FISH) on paraffin-embedded section using IgH/cyclin D1 (CCND1) probe showed that in both cases there was no molecular evidence of t(11;14), finally leading to the diagnosis of CD5-positive MALT lymphoma. Although the present two patients had no recurrence over 34 months after initial diagnosis, careful observation is needed because the clinicopathological significance of MALT lymphoma with this rare phenotype remains obscure.     

Mantle cell lymphoma lacking the t(11;14) translocation: a case report and brief review of the literature

The diagnosis of mantle cell lymphoma (MCL) requires a multifaceted approach with integration of morphology and immunophenotype, supported by cyclin D1 positivity or identification of t(11;14)(q13;q32). Interphase fluorescence in situ hybridisation (FISH) using a dual colour, dual fusion probe strategy for t(11;14) is a rapid test with high sensitivity and specificity for MCL, and is easily performed on routine bone marrow aspirate or peripheral blood specimens. This test has become the method of choice for many pathologists to confirm a diagnosis of MCL. This report describes a case of MCL with a normal (negative) FISH signal pattern for t(11;14) that was found to be cyclin D1 positive by immunohistochemistry in tissue sections. This case illustrates the need for additional testing when the t(11;14) abnormality is not identified but the morphology and immunophenotype are otherwise suggestive of MCL.     

Early lymph node involvement by mantle cell lymphoma limited to the germinal center: report of a case with a novel "follicular in situ" growth pattern

Recently, several reports have described cases of "in situ" mantle cell lymphoma (MCL) in which scattered cyclin D1+ cells were present within the mantle zones of reactive-appearing lymphoid follicles. In this report, we describe an unusual histologic pattern of in situ MCL that was identified in a staging lymph node for colonic adenocarcinoma resected 4 years before a diagnosis of symptomatic MCL. Retrospective immunohistochemical studies showed scattered cyclin D1-expressing cells within otherwise reactive germinal centers but not in the surrounding mantle zones. The presence of early MCL cells limited to reactive germinal centers represents a novel "follicular in situ" growth pattern for MCL, which overlaps morphologically with reactive follicular hyperplasia and follicular lymphoma and which could have implications for MCL pathogenesis.     

Mantle cell lymphoma: recent insights into pathogenesis, clinical variability, and new diagnostic markers

Mantle cell lymphoma (MCL; previously called centrocytic lymphoma or lymphocytic lymphoma of intermediate differentiation) is a distinct subtype of B-cell lymphoma, accounting for approximately 3%-10% of all lymphoma diagnoses. The name refers to the growth pattern in early disease presentation resembling the normal mantle zone that surrounds the germinal center of the B-cell follicle. The hallmark of MCL is the t(11;14)(q13;q32), resulting in aberrant expression of the CCND1 gene and expression of cyclin D1 in the tumor cells. Expression and genomic profiling of MCL have provided new insight into the pathogenesis and will be summarized in this review. Pitfalls in the differential diagnosis versus B-cell chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, cyclin D1-positive diffuse large B-cell lymphoma, hairy cell leukemia, and plasma cell tumors will be discussed, including the usefulness new diagnostic markers SOX11 and CD200. In situ MCL, MCL with an indolent clinical course, and cyclin D1-negative MCL are other topics of this review.     

Lymphomatous polyposis of the gastrointestinal tract, including mantle cell lymphoma, follicular lymphoma and mucosa-associated lymphoid tissue lymphoma

AIMS: Lymphomatous polyposis (LP) is considered to represent mantle cell lymphoma (MCL) of the gastrointestinal (GI) tract. However, a few reports have suggested that some are follicular lymphoma (FL) or mucosa-associated lymphoid tissue (MALT) lymphomas. In this study, we analysed 35 patients and clarified the clinicopathological features of LP. METHODS AND RESULTS: Paraffin-embedded tissue samples were stained immunohistochemically and analysed by tissue-fluorescence in situ hybridization (T-FISH) for IGH/CCND1 (cyclin D1) and IGH/BCL2. The average age of the patients was 58.3 years. Over half of the cases showed gastric, duodenal, small intestinal, ileocaecal and sigmoid colonic lesions (15, 19, 15, 16 and 16 cases, respectively). Phenotypically, cases were classified into three types of MCL (cyclin D1+ CD5+ CD10-) (n=12), FL (cyclin D1- CD5- CD10+) (n=14) and MALT (cyclin D1- CD5- CD10-) (n=9). T-FISH identified 11 of the 11 examined cases with MCLs to have IGH/CCND1, while seven of 10 cases with FL had IGH/BCL2, and none of the MALT cases were positive for IGH/CCND1 or IGH/BCL2. At the study endpoint, five of 12 patients with MCL were dead, two of 14 with FL and one of nine with MALT were dead of other disease. Event-free survival analysis showed significantly poorest outcome in MCL, followed by FL, while MALT was associated with a favourable outcome (P=0.0040). CONCLUSIONS: Our study emphasizes the importance of differentiating MCL, FL and MALT of LP in evaluating prognosis and hence the most suitable therapeutic regimen.     

Immunohistochemical detection of cyclin D1 using optimized conditions is highly specific for mantle cell lymphoma and hairy cell leukemia.

Mantle cell lymphoma (MCL) is more aggressive when compared with other lymphomas composed of small, mature B lymphocytes. Cyclin D1 is overexpressed in MCL as a result of the translocation t(11;14)(q13;q32). Cyclin D1 immunohistochemistry in fixed, paraffin-embedded tissue contributes to the precise and reproducible diagnosis of MCL without the requirement of fresh tissue. However, its use in bone marrow biopsies is not well established. In addition, increased levels of cyclin D1 mRNA have been found in hairy cell leukemia but have not consistently been detected by immunohistochemistry. We used a polyclonal antibody and heat-induced antigen retrieval conditions to evaluate 73 fixed, paraffin-embedded bone marrow, spleen, and lymph node specimens with small B-cell infiltrates, obtained from 55 patients. Cyclin D1 was overexpressed in 13/13 specimens of MCL (usually strong, diffuse reactivity in most tumor cells) and in 14/14 specimens of hairy cell leukemia (usually weak, in a subpopulation of tumor cells). No reactivity was detected in five cases of B-chronic lymphocytic leukemia; five cases of splenic marginal zone lymphoma; six cases of nodal marginal zone cell lymphoma; two cases of gastric marginal zone cell lymphoma; or ten benign lymphoid infiltrates in bone marrow, spleen, or lymph nodes. In summary, although the total number of studied cases is small and a larger series of cases may be required to confirm our data, we present optimized immunohistochemical conditions for cyclin D1 in fixed, paraffin-embedded tissue that can be useful in distinguishing MCL and hairy cell leukemia from other small B-cell neoplasms and reactive lymphoid infiltrates.     

Composite mantle cell and diffuse large B-cell lymphoma: report of two cases

Composite lymphomas are rare and involve the concurrent evolution of 2 distinct lymphoma types within a single organ or tissue. This study describes 2 cases of composite mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL), which has not previously been reported. Each case demonstrated distinct populations of CD20 positive small and large atypical B cells. In both cases, only the small lymphocytes were positive for CD5 and cyclin D1, and fluorescence in situ hybridization (FISH) showed a t(11;14) translocation in the small lymphocytes but not in the large cells. Molecular studies for B-cell clonality showed a possible clonal relationship between the 2 components in one case but not the other. This study describes in detail the morphology, immunophenotype, FISH, and molecular analysis of both components in each case. To the authors' knowledge, this represents the first report of juxtaposition of MCL with DLBCL that does not represent transformation of the mantle cell component.     

Mantle cell lymphoma with in situ or mantle zone growth pattern: a study of five cases and review of literature

We present two rare cases of in situ mantle cell lymphoma (“in situ MCL”) and three cases of MCL with mantle zone growth pattern (MCL-MZGP). The patients include four males and one female, with a median age of 66 years (range, 52 to 86 years). Two present with isolated lymphadenopathy and three with multiple lymphadenopathy. At presentation, the complete blood count (CBC) and serum lactate dehydrogenase (LDH) are normal in all cases. Histologic examination reveals an in situ pattern in two cases and a mantle zone growth pattern in three cases. The staging bone marrow biopsies show minimal involvement by lymphoma in one case and no morphologic evidence of lymphoma in four cases. All cases are positive for cyclin D1, including two with typical MCL phenotype and three with CD5-negativity. Four out of five cases express kappa light chain. FISH study for t(11;14) is performed in three cases, of which one is positive and two are inconclusive. For four patients with a median follow-up of 38 months, three are in clinical remission and one has persistent disease. In conclusion, the “in situ MCL” is associated with incidental finding, indolent clinical course and lower tumor burden. The predominant usage of kappa light chain and frequent CD5-negativity observed in our cases are unusual. We review the clinical and laboratory features of “in situ MCL” cases in the literature.     

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。     

Detection of t(11;14) using interphase molecular cytogenetics in mantle cell lymphoma and atypical chronic lymphocytic leukemia

The chromosomal translocation t(11;14)(q13;q32) fuses the IGH and CCND1 genes and leads to cyclin D1 overexpression. This genetic abnormality is the hallmark of mantle cell lymphoma (MCL), but is also found in some cases of atypical chronic lymphocytic leukemia (CLL), characterized by a poor outcome. For an unequivocal assessment of this specific chromosomal rearrangement on interphase cells, we developed a set of probes for fluorescence in situ hybridization (FISH). Northern blotting was performed for analysis of the cyclin D1 expression in 18 patients. Thirty-eight patients, with either a typical MCL leukemic phase (17 patients) or atypical CLL with an MCL-type immunophenotype, i.e., CD19⁺, CD5⁺, CD23^−/low, CD79b/sIgM(D)⁺⁺, and FMC7⁺ (21 patients), were analyzed by dual-color interphase FISH. We selected an IGH-specific BAC probe (covering the JH and first constant regions) and a commercially available CCND1 probe. An IGH–CCND1 fusion was detected in 28 of the 38 patients (17 typical MCL and 11 cases with CLL). Cyclin D1 was not overexpressed in two patients with typical MCL and an IGH–CCND1 fusion. In view of the poor prognosis associated with MCL and t(11;14)-positive CLL, we conclude that this set of probes is a valuable and reliable tool for a rapid diagnosis of these entities. Genes Chromosomes Cancer 23:175–182, 1998. © 1998 Wiley-Liss, Inc.     

Immunohistochemical analysis of cyclin D1 shows deregulated expression in multiple myeloma with the t(11;14)

The t(11;14)(q13;q32) chromosomal translocation, the hallmark of mantle cell lymphoma (MCL), is recurrently found in multiple myelomas (MM) by means of conventional cytogenetics. Unlike MCL, recent molecular studies of MM-derived cell lines with t(11;14) have indicated that the breakpoints are highly dispersed over the 11q13 region; however, the fact that cyclin D1 is generally overexpressed in these cell lines suggests that this gene is the target of the translocation. To evaluate further the involvement of cyclin D1 in MM, we used immunohistochemistry and fluorescence in situ hybridization to investigate cyclin D1 expression and the presence of chromosome 11 abnormalities in a representative panel of 48 MM patients (40 at diagnosis and 8 at relapse). Cyclin D1 overexpression occurred in 12/48 (25%) of cases; combined immunohistochemistry and fluorescence in situ hybridization analyses in 39 patients showed cyclin D1 positivity in all of the cases (7/7) bearing the t(11;14), in two of the 13 cases with trisomy 11, and in one of the 19 cases with no apparent abnormalities of chromosome 11. Our data indicate that the t(11;14) translocation in MM leads to cyclin D1 overexpression and that immunohistochemical analysis may represent a reliable means of identifying this lesion in MM.     

Classical type and blastoid variant mantle cell lymphoma in the same lymph node: Histology and cytological findings from a touch imprint specimen

Blastoid variant (BV) is one of the aggressive variants of mantle cell lymphoma (MCL). BV‐MCL is defined by its blastic cytomorphology. Previous studies using sequential biopsies in cases with MCL have demonstrated that classical type MCL (C‐MCL) often transforms or relapses as an aggressive variant, but a histopathological transition from C‐MCL to an aggressive MCL variant in the same pathological specimen has been shown in only a limited number of the cases. We present a case of MCL in which a histological transition between C‐MCL and BV‐MCL was observed in the same lymph node. A 53‐year‐old man presented with a submandibular tumor. Touch imprint cytology revealed a monotonous proliferation of large blastic lymphoid cells. Histology revealed a transition between a large lymphoid cell component and small foci of small‐ to medium‐sized cell component within the tumor. Both components were CD5(+), CD10(−), CD20(+), cyclin D1(+), and SOX11(+) on immunohistochemistry. Fluorescent in situ hybridization revealed the translocation of IgH/BCL1 locus. These findings led to a final diagnosis of BV‐MCL with coexistent C‐MCL. The present case suggests the existence of a pathogenetic pathway of MCL from C‐MCL to BV‐MCL. Because it is important to accurately identify BV‐MCL for prognostication, appropriate ancillary diagnostic tools should be used in suspected cases. Diagn. Cytopathol. 2017;45:364–370. © 2016 Wiley Periodicals, Inc.     

Indolent mantle cell lymphoma with nodal involvement and mutated immunoglobulin heavy chain genes

Mantle cell lymphoma (MCL) is typically considered an aggressive but incurable neoplasm composed of cyclin D1+ monoclonal B-cells with a t(11;14)(q13;q32) and usually unmutated immunoglobulin (Ig) genes. Although it has been suggested that a more indolent leukemic disorder exists with the same phenotype and genotype but with mutated Ig genes, others have considered these cases to be variants of chronic lymphocytic leukemia. We present a case of an indolent MCL that was documented with cyclin D1 expression in a lymph node biopsy performed more than 12 years ago. The patient has peripheral blood involvement with a lymphocyte count in the reference range, variable thrombocytopenia, and minimal adenopathy but is otherwise well, never having received any antineoplastic therapy. Study of peripheral blood samples from 2002 revealed a CD5-variable B-cell monoclonal proliferation with a t(11;14)(q13;q32) plus other karyotypic abnormalities, positive fluorescence in situ hybridization studies for the CCND1/IgH translocation, and clonal Ig gene rearrangement with mutated Ig genes (95.7% homology to VH 4-31). The subtle but diagnostic lymph node biopsy in this case helps to further support that an indolent t(11;14) monoclonal lymphocytosis with mutated Ig genes can represent an MCL variant rather than chronic lymphocytic leukemia.     

套细胞淋巴瘤石蜡包埋组织中细胞周期蛋白D1和t(11;14)易位检测的临床病理意义

目的 探讨套细胞淋巴瘤石蜡包埋组织中细胞周期蛋白(cyclin)D1和t(11;14)易位检测的可行性及其诊断和鉴别诊断价值。方法 收集套细胞淋巴瘤36例,对照组小B细胞恶性淋巴瘤71例,均为石蜡包埋组织,运用免疫组织化学方法观察cyclin D1的表达;用半巢式聚合酶链反应(PCR)法检测t(11;14)易位,以看家基因β-肌动蛋白(actin)作为内对照检测DNA质量。结果 (1)36例套细胞淋巴瘤中26例(72．2％)表达cyclin D1,对照组无1例表达。(2)107例标本中101例(94．4％)可检出β-actin DNA表达。36例套细胞淋巴瘤中22例检出t(11;14)易位,对照组无1例检出。去除B-actin和t(11;14)易位均阴性2例,套细胞淋巴瘤中t(11;14)易位检出率为64．7％。(3)36例套细胞淋巴瘤中cyclin D1染色和(或)t(11;14)易位检测阳性病例为29例,总阳性率为80．5％。结论套细胞淋巴瘤石蜡包埋组织中cyclin D1和t(11;14)易位的检测具较高的特异性和可行性,两者的综合应用有助于正确的诊断和鉴别诊断。     

Blastoid and common variants of mantle cell lymphoma exhibit distinct immunophenotypic and interphase FISH features

AIMS: The recognition of blastoid variant (BV) of mantle cell lymphoma (MCL) is based on morphological criteria. Our aim was to analyse 18 MCL cases including four BV-MCL for their clinicopathological features, proliferation index, cyclin D1 and CDK4 expression and interphase fluorescence in-situ hybridization (FISH) pattern. METHODS AND RESULTS: BV-MCL versus common MCL was characterized by a shorter overall duration of response after first-line therapy (11 months versus 28 months) and shorter overall survival (20 months versus 42 months). Interphase FISH showed a t(11;14) fusion pattern in all MCL tested cases. However, the four blastoid cases were characterized by extra copies of CCND1 signals. Using additional probes of chromosomes 11, 18, 21, these signals were shown to be the result of hypotetraploidy and not of a specific amplification of the normal or the translocated CCND1 allele. Moreover, the BV-MCL cases were characterized by a combined high percentage of cells expressing cyclin D1 and/or CDK4 with a proliferation (MIB-1-Ki67) index above 50%. Such features allowed the recognition of areas of large cell transformation in the case of secondary BV-MCL. CONCLUSIONS: Since distinction between BV and common MCL is of clinical relevance, our data underline the need to add phenotypic and cytogenetic criteria to cytomorphology for a better recognition of BV-MCL.     

Mantle cell lymphoma: improved diagnostics using a combined approach of immunohistochemistry and identification of t(11;14)(q13;q32) by polymerase chain reaction and fluorescence in situ hybridization

INTRODUCTION: Mantle cell lymphoma (MCL) is a clinicopathological entity characterized by an aggressive clinical course, morphological features, and overexpression of cyclin D1 due to juxtaposition of the bcl-1 locus (and CCND1 gene coding for the cyclin D1) to the IgH gene. This phenomenon is caused by t(11;14)(q13;q32). The morphological diagnosis of MCL may pose difficulties. Ancillary methods are available to support the diagnosis. PATIENTS AND METHODS: We studied a group of 32 patients with MCL; 24 men and 8 women. The median age at the diagnosis was 64 years. We characterized the investigated group by histology, and to analyze the immunohistochemical (IHC) profile we used a panel of antibodies including anti-cyclin D1. Polymerase chain reaction (PCR) was used to detect the rearrangement of bcl-1/IgH in 26 cases (in 11 patients, the DNA was isolated from frozen tissues or from nucleated cells of bone-marrow aspirate or peripheral blood, in 15 patients we utilized paraffin-embedded material). Dual color fluorescence in situ hybridization (FISH) on interphase nuclei detecting the t(11;14)(q13;q32) was applied in all 32 cases. RESULTS: Cyclin D1 IHC was positive in 29 of 30 cases tested (97%). In six, the result was weak and difficult to rely on to support the diagnosis. PCR revealed the fusion gene in 14 of the 26 cases (54%). The best yield was obtained from fresh and frozen samples (8 of 11 positive). Using FISH, we identified the translocation in all 32 patients, the findings being easily interpretable in 29 patients. In three cases, the intensity of red and green signals was weaker and difficult to read though the co-hybridized signals were identified. The classical pattern of the translocation was observed in 26 patients, while in 3 we found variant patterns suggesting a loss of the V segment of the IgH gene (2x) and a shift in the breakpoint region at chromosome 11 (1x). CONCLUSION: The diagnosis of MCL should be supported by a complex laboratory approach. Interphase FISH seems a useful complementary method to morphology and IHC. It is applicable to various tissues and cells prepared as tissue imprints or histological sections.     

p27(Kip1) immunostaining for the differential diagnosis of small b-cell neoplasms in trephine bone marrow biopsies.

The distinction between mantle cell lymphoma (MCL) and other small B-cell non-Hodgkin lymphomas (NHL) is important because MCL has a more aggressive clinical course. In bone marrow (BM) biopsy specimens, this distinction can be particularly difficult. Although cyclin D1 immunostaining and molecular detection of the t(11;14) translocation are highly specific markers for MCL, they fail to detect a proportion of cases. We have recently described that MCL typically lacks detectable expression of the cyclin-dependent kinase inhibitor p27(kip1) protein by immunostaining, which is expressed at high levels in most small B-cell NHL inversely correlated to the proliferation rate. We therefore examined whether p27(kip1) immunostaining could be a useful adjunct for the differential diagnosis of small B-cell NHL infiltrates in the BM. Trephine BM biopsy specimens of 96 patients, including well-characterized MCL (19 cases), B-cell chronic lymphocytic leukemia (27 cases), follicular lymphoma (18 cases), hairy cell leukemia (22 cases), and marginal zone lymphoma (10 cases) as well as 10 reactive BM, including five with benign lymphoid aggregates were investigated. In addition, the presence of a t(11;14) translocation involving the major translocation cluster was studied by PCR in all MCL. All cases of B-cell chronic lymphocytic leukemia, follicular lymphoma, and marginal zone lymphoma revealed a strong p27(kip1) nuclear staining in the majority of neoplastic cells. Fourteen (78%) cases of MCL were p27(kip1)-negative in the tumor cells, whereas four cases revealed a weak nuclear positivity. Seventeen (77%) cases of hairy cell leukemia were also either completely negative for p27(kip1) or showed a faint positive staining in a minority of the neoplastic cells. Nine of 19 cases (47%) of MCL showed a bcl1 rearrangement involving the major translocation cluster region. These findings demonstrate that p27(kip1) immunostaining is a valuable additional marker for the differential diagnosis of small B-cell NHL infiltrates in BM biopsies. The reduction or lack of p27(kip1) protein expression in MCL, as well as in hairy cell leukemia, might be an important event in the pathogenesis of these disorders.