Epstein-Barr virus is rarely associated with diffuse large B cell lymphoma in Taiwan and carries a trend for a shorter median survival time.

AIMS: Epstein-Barr virus (EBV)-positive diffuse large B cell lymphoma (DLBCL) of the elderly is characterised by frequent extranodal involvement, a morphological spectrum from polymorphous to monomorphous and a poor prognosis. The frequency is higher in Japan and Korea but lower in the West, while the status in Taiwan has not been reported yet. METHODS: We conducted a retrospective study of DLBCL in a single institute in Taiwan by immunohistochemistry and in situ hybridisation for EBV. RESULTS: Of the 424 consecutive DLBCL cases, 332 cases were studied for EBV. 15 (4.5%) were EBV-positive and 13 (3.9%) fulfil WHO criteria of EBV-positive DLBCL of the elderly with a median age of 75. Of these 15 cases, extranodal presentation occurred in 11 (73%) patients with predominance in the gastrointestinal tract and 6 (40%) were of germinal centre B cell phenotype. There was no difference between EBV-positive and -negative patients in terms of age, gender, nodal versus extranodal presentation, and immunophenotypical profile. EBV-positive patients showed a trend for a shorter median survival time (5.0 vs 39.3 months; p=0.058). Of all DLBCL patients, multivariable analysis revealed a significantly worse overall survival for patients older than 50 (p=0.001) and for those with bcl-6-negative tumours (p=0.003) but not with other clinicopathological factors including EBV status. CONCLUSIONS: EBV-positive DLBCL of the elderly is relatively rare in Taiwan, with an incidence intermediate between Japan/Korea and the West. Further studies are warranted to clarify the association of EBV and the clinicopathological features and the prognostic significance in patients with DLBCL.

Follicular lymphoma in Taiwan: a low frequency of t(14;18), with grade 3A tumours more closely related to grade 3B than to low-grade tumours.

AIMS: To investigate t(14;18)/IGH-BCL2 in follicular lymphoma (FL) cases from Taiwan. METHODS AND RESULTS: We retrospectively studied 93 consecutive cases, using immunohistochemistry and fluorescence in-situ hybridization (FISH). Fifty-nine (63%) tumours were low-grade (LG) and 34 (37%) were high-grade (HG; 24% FL3A and 13% FL3B). FISH showed IGH, BCL2 and BCL6 rearrangements in 59%, 47% and 11% of cases, respectively, and MYC rearrangement in 5% of FL3A tumours and 25% of FL3B tumours. The translocation partner of all BCL2 rearrangements was IGH, with IGH-BCL2 fusion in 63% of LG tumours and 18% of HG tumours. LG tumours were enriched with a CD10+/bcl-2+/MUM1- phenotype, and were frequently associated with BCL2 rearrangement but less commonly with BCL6 rearrangement. FL3A tumours were more closely related to FL3B tumours than to LG tumours in immunophenotype and genetic aberrations. There was no statistically significant difference between grade 1 and two tumours, between FL3A and FL3B tumours or between nodal and extranodal tumours in immunophenotypic or FISH findings. The cumulative survival rate was higher in LG FL patients with IGH-BCL2 translocation than in those without rearrangement. CONCLUSIONS: In Taiwan, FL3A tumours were more closely related to FL3B tumours than to LG tumours, and a literature review showed that the frequency of t(14;18)/IGH-BCL2 in FL in Taiwan is among the lowest in the world.

SOX11 is useful in differentiating cyclin D1-positive diffuse large B-cell lymphoma from mantle cell lymphoma.

AIMS: To characterize the frequency and clinicopathological features of cyclin D1-positive diffuse large B-cell lymphoma (DLBCL) and the usefulness of SOX11 in the differential diagnosis from mantle cell lymphoma (MCL). METHODS AND RESULTS: We retrospectively stained 206 consecutive DLBCLs for cyclin D1, and identified three (1.5%) positive cases, comprising two in the elderly with necrosis, and a third with a starry-sky pattern. All three cases shared the same non-germinal centre B-cell (non-GCB) phenotype [CD5-/CD10-/bcl-6+/MUM1+/SOX11-], Epstein-Barr virus (EBV) negativity, and absence of CCND1 aberrations by fluorescence in-situ hybridization. The third case showed both BCL6 and MYC rearrangements: a double-hit lymphoma. In the same period there were 22 MCLs, all expressing cyclin D1, with 89% cases expressing SOX11, a frequency that is statistically different from cyclin D1-positive DLBCL. Notably, we identified a pleomorphic MCL initially misdiagnosed as DLBCL. A separate cohort of 98 DLBCL cases was negative for SOX11, with only one case expressing cyclin D1 with a GCB phenotype (CD10+/bcl-6+/MUM1-). The two patients with tumour necrosis rapidly died of disease. The other two were in complete remission after immunochemotherapy. CONCLUSION: Cyclin D1-positive DLBCLs are rare, and they are negative for SOX11 or CCND1 aberration. SOX11 is useful in differentiating cyclin D1-positive DLBCL from MCL.

Therapy-related acute lymphoblastic leukemia with t(4;11)(q21;q23) masqueraded as marrow lymphocytosis in a patient with breast cancer.

Therapy-related acute leukemia develops in patients after chemotherapy and/or radiotherapy for a prior cancer, and most cases are acute myeloid leukemia with a much lower frequency of acute lymphoblastic leukemia (ALL). One unique feature of these therapy-related ALL (t-ALL) is an increased incidence of chromosome band 11q23 aberrations as compared with de novo ALL. In adult female patients, breast cancer is the most common primary cancer. Herein, we report the case of a 49-year-old Taiwanese lady who developed t-ALL with t(4;11)(q21;q23) 16 months after cyclophosphamide, epirubicin, and 5-fluorouracil chemotherapy for her breast cancer. The unusual feature is that the t-ALL was heralded 4 months ago by marrow lymphocytosis comprising atypical small lymphocytes with condensed chromatin mimicking a B-cell chronic lymphoproliferative disorder. Retrospective studies using additional antibodies for immunophenotyping and PCR-based clonality study for immunoglobulin gene rearrangement showed that these atypical small lymphocytes shared similar features with the leukemic blasts at the frank leukemic stage. Our results suggest that these atypical small lymphocytes are lymphoblasts in disguise and that the clinicopathological correlations with ancillary pathological studies are important to reach a definitive diagnosis of such an unusual case.

A parallel comparison of T-cell clonality assessment between an in-house PCR assay and the BIOMED-2 assay leading to an efficient and cost-effective strategy.

AIMS: Diagnosis of T-cell lymphoproliferation is sometimes challenging, and in certain instances pathologists rely heavily on the clonality assessment results of T-cell receptor (TCR) gene rearrangement (TCR-GR). Many investigators have designed various in-house primer sets for PCR-based study targeting different loci of TCR genes. In recent years, the commercial BIOMED-2 protocols have become available. The in-house primers are very cheap while the BIOMED-2 primers are expensive. This parallel study aimed to compare the sensitivity of the in-house TCRG primers (two reactions) and the BIOMED-2 TCR primers (six reactions) in an attempt to develop a sensitive and cost-effective strategy for TCR-GR assessment. METHODS: PCR-based analysis was performed on 69 samples of T-lineage neoplasms including 60 formalin-fixed paraffin-embedded (FFPE) tissues, 5 samples from peripheral blood (PB) and 4 samples from bone marrow (BM) aspirate. RESULTS: Forty-seven (78%) FFPE and all PB or BM aspirate samples yielded control DNA products suitable for clonality assessment including 4 precursor and 50 mature T-cell neoplasms. The detection rates of clonal TCR-GR were 63% (34/54) by the two in-house TCRG primers, 85% (46/54) by all six BIOMED-2 reactions, 91% (49/54) by combining the in-house and BIOMED-2 TCRG reactions and 94% (51/54) by combining the in-house and all BIOMED-2 reactions. By using the in-house and BIOMED-2 TCRG reactions with a total of four tubes, clonal TCR-GR was detected in 91% of the cases. The reagent cost for this combination was one-third of that for the six BIOMED-2 reactions and the detection rate was also higher than the latter alone (91% vs 85%). CONCLUSIONS: As the in-house primers were custom made and are much cheaper than the commercial kits, the authors concluded that this four-tube strategy was cost-effective and efficient for TCR-GR clonality assessment.

Hepatitis C virus infection is significantly associated with malignant lymphoma in Taiwan, particularly with nodal and splenic marginal zone lymphomas.

AIMS: Hepatitis C virus (HCV) is a hepatotropic and lymphotropic RNA virus causally linked to lymphoma with a strong geographic variation. The aim of this study was to investigate the association of HCV and lymphoma in Taiwan, in which HCV is endemic. METHODS: Patients diagnosed with lymphoma from January 2004 to December 2008 were investigated for serum anti-HCV, and the infection rate was compared with that in healthy controls. Various lymphoma types were investigated for HCV infection. Immunohistochemistry was performed for HCV non-structural (NS)3 protein, and genotyping was performed by reverse transcriptase PCR. RESULTS: Thirty-eight (11.0%) of 346 patients with lymphoma were positive for anti-HCV, as compared with 15 (1.8%) of 824 healthy controls (p<0.001, chi(2) test) with an age-adjusted and sex-adjusted OR of 4.57 (95% CI 2.41 to 8.68). Only nodal (five of eight cases) and splenic (two of two cases) marginal zone lymphomas (MZLs) as a group were significantly associated with HCV, as compared with mucosa-associated lymphoid tissue (MALT) lymphomas (1 of 15; p=0.002, Fisher's exact test). All 26 anti-HCV-positive cases stained for HCV-NS3 were negative. The most common genotypes were 1b (22%) and 2a (56%), with no statistical difference from 203 patients with HCV-related chronic liver disease. CONCLUSIONS: The incidence of HCV infection among lymphoma patients in Taiwan was significantly higher than that for healthy controls. Furthermore, non-MALT (nodal and splenic) MZL was the only group significantly associated with HCV. A larger national study is warranted to re-confirm our findings and to elucidate if any particular HCV genotypes were related to the pathogenesis of lymphoma.

Multicentric primary intestinal EBV-positive diffuse large B cell lymphoma of the elderly presenting with perforation.

Epstein-Barr virus (EBV)-positive diffuse large B cell lymphoma (DLBCL) of the elderly is a recently defined subgroup of DLBCL in the 2008 WHO classification of lymphoid neoplasms. It is characterized by an EBV-positive clonal B cell lymphoproliferation that occurs in patients aged over 50 years without any known immunodeficiency. This disease shows frequent extranodal involvement, a morphological spectrum from polymorphous to monomorphic large cells, and a poor prognosis. We reported the case of a 75-year-old male who had a primary intestinal multicentric EBV(+) DLBCL and presented with perforation, the first multicentric case in the literature. The monomorphic tumor cells exhibited extensive necrosis and expressed CD20, CD138, and EBNA2 with diffuse positivity for EBV by in situ hybridization. Quantitative real-time PCR revealed a low serum EBV viral load. The patient passed away in 10 days. His poor outcome was probably due to perforation, age over 70, the presence of B symptoms, poor general condition, and high performance score.

Angioimmunoblastic T-cell lymphoma of cytotoxic T-cell phenotype containing a large B-cell proliferation with an undersized B-cell clonal product.

Angioimmunoblastic T-cell lymphoma is a nodal peripheral T-cell lymphoma considered to be derived from CD4 follicular helper T cells. It is characterized by the proliferation of arborizing vessels, hyperplastic follicular dendritic cells, and a polymorphous lymphoid infiltrate including large B immunoblasts, which could be polyclonal, oligoclonal, or monoclonal. The polymerase chain reaction-based clonality study of lymphoproliferations is increasingly popular in the diagnostic workup. With the commercially available Biomed-2 protocols, lymphoproliferations with amplicons in the expected size ranges are considered clonal, whereas clonal products outside the expected ranges are extremely rare. We presented the case of a 60-year-old male patient with angioimmunoblastic T-cell lymphoma, in which the neoplastic T cells expressed CD8, bcl-6, and programmed death-1. Furthermore, there was a proliferation of large B cells in this tumor. The results of T-cell receptor gene rearrangement study using the Biomed-2 protocols showed clonal rearrangement with amplicons falling within the expected size ranges. Interestingly, the size of the amplicons detected by the Biomed-2 immunoglobulin heavy chain gene (IgH) rearrangement using FR2/JH primers was around 240 bp, slightly smaller than the expected size ranges. Through cloning, sequencing, and BLAST searches, we confirmed that the FR2/JH amplicon was derived from the IgH rearrangement with a deletion of a short segment. Our case illustrates that polymerase chain reaction amplicons outside the expected size ranges may still be clonal products.

Primary cutaneous diffuse large B cell lymphoma relapsed solely as a huge lung tumor mimicking a primary pulmonary lymphoma.

Primary cutaneous large B cell lymphoma, leg type (PCLBCL-leg) has recently been identified and recognized as a specific entity. Patients with PCLBCL-leg have a higher relapse rate and a poorer prognosis than the other types of primary cutaneous B cell lymphomas, and disease relapse is confined to the skin in the majority of cases with rare exclusive extracutaneous progression. The late occurrence of lymphoma in patients with a prior history of lymphoma may represent a relapse/progression or a distinct tumor unrelated to the original one. The distinction is of important clinical and therapeutic implications. Here, we report the case of a 90-year-old lady with a history of PCLBCL-leg in complete remission after radiotherapy that developed a huge, solitary pulmonary lymphoma without lymphadenopathy 14 months later. The latter was initially considered as stage IE primary pulmonary lymphoma and was treated with combination chemotherapy resulting in complete remission. Retrospective pathologic review and B cell clonality study revealed that the pulmonary tumor was a diffuse large B cell lymphoma of the same clonal origin as the PCLBCL-leg. This case is unique in the exclusive pulmonary relapse and illustrates the importance of expert pathological review and molecular study in the management of lymphoma patients with unusual clinical features.