Cytogenetic analysis of de novo CD5-positive diffuse large B-cell lymphoma

Aims: De novo CD5‐positive diffuse large B cell lymphoma (CD5+DLBL) is a subtype of DLBL with poor clinical outcome. To investigate the cytogenetic pathogenesis of CD5+DLBL, we analyzed the chromosomal findings of 18 patients with CD5+DLBL. Methods: Tumor cells were cultured and metaphase was captured by colchicine exposure. Using trypsin‐Giemsa banding, the chromosomes were analyzed according to the International System for Human Cytogenetic Nomenclature. Results: Metaphase was acquired from 12 patients. Normal karyotypes were seen in two patients and abnormal karyotypes in the remaining 10. The numbers of chromosomes ranged from 45 to 90. Gain, loss and rearrangements of various chromosomes were seen. Frequent breakpoints were located at chromosome 1 band p13, 3q27, 6q13, 7q32, 14q32, 18q21 and 19q13. There was no diagnosis‐specific abnormality. A relationship between chromosomal findings and clinical outcomes such as involved site, relapse or survival, was not observed. Conclusion: Since previous and the present studies on the chromosomal analysis of CD5+DLBL are also contradictory, more detailed comprehensive genetic analysis appears to be needed to elucidate the biological mechanisms of CD5+DLBL.     

Clinicopathologic characteristics and treatment outcome of the addition of rituximab to chemotherapy for CD5-positive in comparison with CD5-negative diffuse large B-cell lymphoma

Background: CD5-positive (CD5+) diffuse large B-cell lymphoma (DLBCL) comprises ∼10% of DLBCLs, and it is associated with poor prognosis. The clinicopathologic characteristics and prognosis of CD5-negative (CD5-) DLBCL and CD5+ DLBCL were compared.Patients and methods: The subjects were 607 DLBCL patients in whom cell surface markers could be analyzed, among 930 consecutive patients registered in the Adult Lymphoma Treatment Study Group between 1998 and 2008.Results: In all, 102 patients (16.8%) had CD5+ DLBCL. Compared with CD5- DLBCL, CD5+ DLBCL was more closely associated with elevated serum lactate dehydrogenase level, advanced stage, poor performance status, extranodal sites, CD10-, BCL-2+, MUM1+, and nongerminal center B-cell type. The 5-year overall survival (OS) rates of CD5+ DLBCL (n = 102) and CD5- DLBCL (n = 505) were 55% and 65%, respectively (P = 0.032), with 5-year progression-free survival (PFS) rates of 52% and 61%, respectively (P = 0.041). In the CD5+ DLBCL patients, the addition of rituximab to chemotherapy significantly improved PFS (4-year PFS, 47.4% versus 62.5%), but not OS (4-year OS, 57.8% versus 63.5%).Conclusions: For CD5+ DLBCL, the addition of rituximab to chemotherapy significantly improved the PFS, but not OS. Therefore, it is thought that a new treatment strategy is necessary for CD5+ DLBCL.     

De novo CD5-positive diffuse large B-cell lymphoma of the skin arising in chronic limb lymphedema

We report the case of a 79-year-old woman with a longstanding lymphedema of the right arm who developed a skin lymphoma involving the right wrist area. Microscopically, the lesion was composed of numerous centroblasts infiltrating both the dermis and the subcutaneous tissue. Phenotypic investigations showed expression of CD20, CD79a, and bcl-2 protein by neoplastic cells. In addition, these cells were CD5 positive. No expression of anaplastic large cell lymphoma kinase (ALK), CD10, CD23, CD30, CD43, bcl-6, cyclin D1, p53 or p16INK4a could be seen. Polymerase chain reaction (PCR) analysis demonstrated a clonal rearrangement of the genes coding for the kappa light chain of the immunoglobulin (Ig). No rearrangement of the genes coding for the Ig heavy chain, t(14;18) or t(11;14) chromosome translocations, or Epstein-Barr virus (EBV) genomic sequences could be found. The tumor was classified as stage IE and was first cured by complete surgical excision. Nineteen months later, a recurrence was noted in the right elbow area. This study further illustrates that lymphoma of the skin may complicate chronic limb lymphedema. Like most of the previously reported cases, this neoplasm belonged to the category of diffuse large B-cell lymphoma. However, it showed CD5 expression as a singular feature.     

T-cell-rich B-cell lymphoma: a clinicopathologic study of 21 cases and comparison with 43 cases of diffuse large B-cell lymphoma

Clinicopathologic features of 21 patients with T-cell-rich B-cell lymphoma (TCRBCL) were reviewed and compared to 43 patients with diffuse large B-cell lymphoma (DLBCL) to determine if there were distinguishing clinical characteristics and differences in response or survival to CHOP therapy. For the diagnosis of TCRBCL, the current WHO criteria was used. In all of our cases, the majority of cells are non-neoplastic T cells and <10% large neoplastic B cells are present. The initial pathologic diagnosis was nodular lymphocyte predominant Hodgkin's lymphoma (NLPHL) in two cases. Patients with TCRBCL were significantly younger (median: 46 years) and had a significantly higher incidence of B symptoms (62%), hepatomegaly (33%) and marrow infiltration (33%) at presentation when compared to DLBCL (P<0.03). The CR rate after treatment was 48% for TCRBCL patients versus 79% for the DLBCL (P<0.003). Although the CR rates in between the two groups are significant, the difference in 3 years survival rates in each CR groups was insignificant (80% versus 77%). The overall survival time in the two groups was 17 months. Event-free survival time in TCRBCL was 12 months, compared with 17 months in the DLBCL (P>0.05). The frequency of patients with TCRBCL achieving CR was 52.6% whereas that of patients with DLBCL was 79% (P<0.003). The TCRBCL 3 years event-free survival 48% and overall survival 64% were 63 and 72% for DLBCL, respectively.     

Comparison of genetic aberrations in CD10+ diffused large B-cell lymphoma and follicular lymphoma by comparative genomic hybridization and tissue-fluorescence in situ hybridization

CD10 is one of the hallmarks of germinal center B-cells where follicular lymphomas (FL) originate. It has not been clearly established, however, whether CD10(+) diffuse B-cell lymphomas (DLBCL) are genetically similar to FL. We therefore examined 19 CD10(+) DLBCL and 40 FL by means of comparative genomic hybridization (CGH) and tissue-fluorescence in situ hybridization (T-FISH). Chromosomal imbalance was more frequently detected in CD10(+) DLBCLs (19/19) than in FLs (24/40). Significant differences were found in eight frequently imbalanced regions, namely those with gains of chromosomes 7q and 12 and those with losses of chromosomes 1p, 4p, 6q, 15q, 16p and 17. Amplification of the 3q region where BCL6 is located is reported to occur frequently in DLBCL, but it was only found in one of the 19 CD10(+) DLBCL cases we examined. The involvement of t(14;18) in CD10(+)+ DLBCL (31%) and in FL (73%) was significantly different (P = 0.0064). The CGH pattern of CD10(+) DLBCL with t(14;18) was also different from that of FL with t(14;18). Taken together, our results indicate that CD10(+) DLBCL constitutes a unique subtype entity with genetic characteristics significantly different from those of FL and DLBCL.     

具有微绒毛特征的CD30阳性弥漫大B细胞淋巴瘤一例临床病理特征

目的 讨论具有微绒毛特征的CD30+弥漫大B细胞淋巴瘤的临床、病理特征,提高对该病的认识与诊断水平.方法 对1例76岁女性的颈部肿大的淋巴结进行组织学、免疫组织化学、EBER原位杂交和电子显微镜观察,并复习相关文献.结果 组织病理学示,淋巴结正常结构消失,形态单一的、胞质丰富的异型细胞呈片状增生,部分区域可见异型细胞淋巴窦内生长模式.免疫组织化学示,异型细胞CD20+、CD79a+、PAX-5+、CD10+、bcl-6+、MUM-1+,40％肿瘤细胞CD30+,80％肿瘤细胞Ki-67+,肿瘤细胞不表达CK、CD3、CD5、CD15、CD56、EMA、bcl-2.电子显微镜观察,瘤细胞体积大,胞质丰富,瘤细胞表面大量微绒毛,微绒毛长短不一,粗细较均匀,少数有分枝.结论 微绒毛淋巴瘤是一类具有独特形态学改变、免疫组织化学表型和超微结构的淋巴瘤.     

Analysis of the immunoglobulin heavy chain gene variable region of CD5-positive and -negative diffuse large B cell lymphoma.

We analyzed nucleotide sequence and intraclonal diversity of the rearranged immunoglobulin heavy chain gene variable region (VH gene) of CD5+ and CD5- diffuse large B cell lymphoma (DLBCL) to clarify the cell origin of de novo CD5+ DLBCL. Ten cases of CD5+ DLBCL and 29 cases of CD5- DLBCL were analyzed. The frequencies of somatic mutation were 0.7 to 12.9% (average, 6.2%) in CD5+ DLBCL and 2.0 to 25.9% (average, 11.1%) in CD5- DLBCL. The ongoing mutation rate was estimated from the number of further single base-substitutions, expressed as a percentage of the total number of nucleotides in 10 cloned PCR products for each case (%). The averages of the ongoing mutation rate of CD5+ DLBCL (four cases) and CD5 DLBCL (seven cases) were 0.051% and 0.197%, respectively. The rate of CD5+ DLBCL was significantly lower than that of CD5- DLBCL (t-test, P = 0.024). These data may indicate that the cell origin of CD5+ DLBCL is different from that of CD5- DLBCL. CD5 is not an activated antigen in DLBCL, but a specific marker of the B1 subset of the B cells, and de novo CD5+ DLBCL may therefore be derived from this unique subset.     

High resolution array comparative genomic hybridization identifies copy number alterations in diffuse large B-cell lymphoma that predict response to immuno-chemotherapy

Despite recent attempts at sub-categorization, including gene expression profiling into prognostically different groups of "germinal center B-cell type" and "activated B-cell type," diffuse large B-cell lymphoma (DLBCL) remains a biologically heterogenous tumor with no clear prognostic biomarkers to guide therapy. Whole genome, high resolution array comparative genomic hybridization (aCGH) was performed on four cases of chemoresistant DLBCL and four cases of chemo-responsive DLBCL to identify genetic differences that may correlate with response to rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) therapy. Array CGH analysis identified seven DNA copy number alteration (CNA) regions exclusive to the chemoresistant group, consisting of amplifications at 1p36.13, 1q42.3, 3p21.31, 7q11.23, and 16p13.3, as well as loss at 9p21.3 and 14p21.31. Copy number loss of the tumor suppressor genes CDKN2A (p16, p14) and CDKN2B (p15) at 9p21.3 was validated by fluorescence in situ hybridization and immunohistochemistry as independent techniques. In the chemo-sensitive group, 12 CNAs were detected consisting of segment gains on 1p36.11, 1p36.22, 2q11.2, 8q24.3, 12p13.33, and 22q13.2, as well as segment loss on 6p21.32. RUNX3, a tumor suppressor gene located on 1p36.11 and MTHFR, which encodes for the enzyme methylenetetrahydrofolate reductase, located on 1p36.22, are the only known genes in this group associated with lymphoma. Whole genome aCGH analysis has detected copy number alterations exclusive to either chemoresistant or chemoresponsive DLBCL that may represent consistent clonal changes predictive for prognosis and outcome of chemotherapy.     

弥漫大B细胞淋巴瘤CD5表达与临床病理特征及疗效关系的研究

目的:检测CD5蛋白在弥漫大B细胞淋巴瘤(DLBCL)的表达情况,并探讨CD5表达与临床病理特征及疗效的关系。方法:采用免疫组化方法(ElivisionTM法)检测232例患者中CD5蛋白的表达,回顾性研究CD5阳性表达DLBCL与CD5阴性表达DLBCL患者的临床病理特征及化疗效果,运用χ2检验比较两者差异。结果:232例DLBCL患者中,CD5阳性表达率19.8%(46/232)。DLBCL中,CD5表达与年龄(χ2=61.061,P=0.001)、性别(χ2=92.115,P=0.001)、分期(χ2=47.508,P=0.001)、KPS评分(χ2=42.178,P=0.001)、B症状(发热、乏力、盗汗和消瘦,χ2=56.344,P=0.001)、血清LDH水平(χ2=33.341,P=0.001)以及侵犯部位(χ2=78.123,P=0.001)相关;而与血清β2-MG水平无关,χ2=2.495,P=0.138。46例CD5表达阳性的患者中,6例化疗有效,有效率为13.0%(6/46);而186例CD5表达阴性DLBCL患者中,131例患者化疗有效,有效率为70.4%(131/186)。两者比较差异有统计学意义,χ2=50.227,P<0.05。结论:CD5阳性表达的DLBCL好发于老年女性,侵袭性高;CD5阳性表达DLBCL患者比CD5阴性表达的患者疗效差。     

CD5-positive diffuse large B-cell lymphoma: a retrospective study in 337 patients treated by chemotherapy with or without rituximab

BackgroundCD5-positive (CD5+) diffuse large B-cell lymphoma (DLBCL) shows poor prognosis and frequent central nervous system (CNS) relapses under anthracycline-containing chemotherapy. The aim of this study was to determine the prognosis and CNS relapse incidence of CD5+ DLBCL in the rituximab era.Patients and methodsWe analyzed 337 patients with CD5+ DLBCL who received chemotherapy with (R-chemotherapy group; n = 184) or without (chemotherapy group; n = 153) rituximab.ResultsNo significant difference was found in clinical background comparisons between the two groups. In the R-chemotherapy group, 60% of the patients were older than 65 years at diagnosis. Both the complete response rate and overall survival (OS) were significantly better in the R-chemotherapy group (P = 0.0003 and P = 0.002, respectively). Multivariate analysis confirmed that chemotherapy without rituximab was associated with unfavorable OS. However, the probability of CNS relapse did not differ between the two groups (P = 0.89). The CNS relapse was strongly associated with short OS (P < 0.0001). In the R-chemotherapy group, 83% of patients who experienced CNS relapse had parenchymal disease.ConclusionsOur results indicate that rituximab improves the OS of patients with CD5+ DLBCL but does not decrease the CNS relapse rate. More effective treatments with CNS prophylaxis are needed for CD5+ DLBCL patients.     

Lenalidomide combined with R-GDP in a patient with refractory CD5-positive diffuse large B-cell lymphoma: A promising response and review

CD5-positive (CD5+) diffuse large B-cell lymphoma (DLBCL) is associated with poor survival compared with CD5-negative DLBCL. The clinical characteristics of CD5+ DLBCL are different from both CD5-negative DLBCL and other CD5+ B cell lymphomas. There is currently no promising chemotherapy for CD5+ DLBCL. Herein, we report a 49-year-old Asian male with refractory CD5+ DLBCL. He complained of aggravated abdominal pain and weight loss. Computed tomography scan revealed abdominal masses, widespread lymphadenopathy, splenomegaly, and intussusception of the ileocecal junction with bowel wall thickening. Core needle aspiration biopsy of an abdominal mass was performed and immunohistochemistry revealed DLBCL of nongerminal center type. In this report, the dose-intensified R-Hyper CVAD (A) regimen as salvage therapy was introduced but failed to result in substantial improvement over the initially standard R-CHOP regimen. Next, the R-GDP regimen was administered as second-line treatment, but only resulted in a partial response. However, the addition of lenalidomide to R-GDP (R2-GDP) resulted in complete remission. The clinical features, pathogenesis, and possible mechanism of action of lenalidomide in CD5+ DLBCL have been described in the literature. The results of the present case report and literature searches indicate that CD5+ DLBCL may share a common pathway with activated B-cell like (ABC) DLBCL as determined by gene expression profiling. Lenalidomide is expected to induce favorable responses in patients with CD5+ DLBCL.     

ALK-positive diffuse large B-cell lymphoma: report of three cases

Diffuse large B-cell lymphoma positive for anaplastic lymphoma kinase (ALK(+) DLBCL) is a rare variant of diffuse large B-cell lymphoma, with characteristic morphological, immunohistochemical and cytogenetic features. Only 34 cases of ALK-positive diffuse large B-cell lymphoma have so far been reported in the literature. We examined three new cases, which showed similar characteristics to previously reported cases, but with peculiar nuclear-membrane staining for ALK protein in one patient and a 5'-ALK gene deletion in another. All of them had stage IV disease at initial presentation, with poor outcomes. The tumour cells showed immunoblastic/plasmablastic histology and were positive for ALK and Oct2, but negative for CD3, CD20, CD79a, CD30 and PAX5. The staining pattern of ALK protein was cytoplasmic in two patients and associated with the nuclear membrane in one patient. Fluorescence in situ hybridization (FISH) analysis using the ALK break-apart probe revealed ALK gene rearrangements in all three patients, with a 5'-ALK gene deletion in one patient. These three cases suggest that different types of cytogenetic aberrations may involve the ALK gene in ALK-positive diffuse large B-cell lymphoma leading to peculiar immunohistochemical staining patterns.     

CD30-positive EBV-associated diffuse large B-cell lymphoma occurring after immunosuppressive therapy for T-cell prolymphocytic leukemia

We describe the case of a 64-year-old man who developed diffuse large B-cell lymphoma (DLBCL) in less than a year after he was diagnosed and treated for T-cell prolymphocytic leukemia (T-PLL). At the time of diagnosis of T-PLL he had a white blood cell count (WBC) of 38.2×10(9)/L and only few small lymph nodes were identified on physical examination. Hepatosplenomegaly or skin lesions were not present. Peripheral blood examination was remarkable for 91% circulating prolymphocytes, which by flow cytometry immunophenotypic analysis were CD2, CD3, CD5, and CD7 positive and coexpressed CD4 and CD8 (absolute number, 33.4×10(9)/L). T-cell receptor (TCR) β and γ genes rearrangements were identified by polymerase chain reaction (PCR). The patient underwent chemotherapy, but did not completely achieve cytogenetic remission. Nine months after his diagnosis of T-PLL, he underwent surgical excision of a new 7 cm left inguinal mass, and was diagnosed with CD30 positive Epstein-Barr Virus (EBV)-associated DLBCL.     

NLRC5在弥漫性大B细胞淋巴瘤中的作用机制

弥漫性大B细胞淋巴瘤(DLBCL)是我国最常见的恶性淋巴瘤,大多首发自淋巴结.研究发现,核苷酸结合寡聚化结构域样受体(NLR)家族成员NLRC5在淋巴系细胞中表达明显,其可能是淋巴细胞肿瘤发生的重要基础.并且NLRC5能阻断信号传导途径中的核心组分核转录因子-κB(NF-κB),影响肿瘤发生发展.同时NF-κB的持续活化是DLBCL细胞生存的必要条件.因此,NLRC5很可能具有抑制过度炎症反应,进而抑制DLBCL的功能,其有望成为DLBCL免疫疗法的新靶点.     

9例原发睾丸弥漫大B细胞淋巴瘤临床特点及预后分析

目的 探讨原发睾丸弥漫大B细胞淋巴瘤(primary testicular diffuse large B-cell lymphoma,PTDLBCL)的临床床特征、治疗及预后.方法 回顾性分析我院2013年1月至2016年12月确诊的9例PTDLBCL患者的临床资料.结果 9例患者中位发病年龄为64岁(50～72岁),以右侧睾丸受累为主;Ann Arbor分期以早期为主,其中Ⅰ期6例,Ⅲ期1例,Ⅳ期2例;细胞来源以非生发中心来源为主,非生发中心B细胞来源7例,生发中心B细胞来源2例;肿瘤增殖指数Ki-67偏高,Ki-67中位数为80％.治疗以手术加术后辅助化疗为主,其中单纯手术治疗1例,手术联合CHOP方案为主的化疗8例.获完全缓解6例,部分缓解2例,疾病进展1例.随访时间5～34个月,存活6例,死亡3例.结论 原发睾丸弥漫大B细胞淋巴瘤是一组罕见的疾病,复发风险高,一线治疗方案推荐患侧睾丸根治性切除术后联合R-CHOP方案化疗,同时对对侧睾丸及受累野进行放疗及预防性甲氨蝶呤鞘内化疗.     

Expression profiling analysis of the CD5+ diffuse large B-cell lymphoma subgroup: development of a CD5 signature

Diffuse large B-cell lymphoma (DLBCL) accounts for 30% of non-Hodgkin's lymphomas and is known to comprise heterogeneous groups. We previously reported that CD5+ DLBCL is a clinically distinct subgroup of these tumors that is associated with poor prognosis. In our current study, we have used gene expression profiling technology in an attempt to identify new markers and to further characterize the biological features of CD5+ DLBCL. Candidate genes, which showed the greatest difference in expression between 22 CD5+ and 26 CD5- DLBCL cases, were selected from our screening and subjected to clustering analysis. This resulted in identification of a specific mRNA profile (a CD5 signature) for CD5+ DLBCL. The CD5 signature included downregulated extracellular matrix genes such as POSTN, SPARC, COL1A1, COL3A1, CTSK, MMP9 and LAMB3, and comprised upregulated genes including TRPM4. We tested this CD5 signature for its potential use as a relevant marker for CD5+ DLBCL and found that it did indeed recognize this subgroup. The tumors identified by the CD5 signature contained most of the CD5+ DLBCL cases and some CD5- DLBCL cases. Moreover, the subgroup of cases with this CD5 signature showed a poorer prognosis. The subsequent application of the CD5 signature to the analysis of an independent series of DLBCL microarray data resulted in identification of a subgroup of DLBCL cases with a similar clinical outcome, further suggesting that the CD5 signature can be used as a clinically relevant marker of this disease.     

微小RNA与弥漫大B细胞淋巴瘤

微小RNA（miRNA）是一类小分子非编码RNA,通过与目标mRNA互补序列结合,在转录水平调控基因的表达。研究发现,包括弥漫大B细胞淋巴瘤在内的不同恶性肿瘤中存在特定的miRNA表达谱。因此,体液、组织标本中miRNA的表达将有望成为评估弥漫大B细胞淋巴瘤的新指标。     

The characteristics of Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma: comparison between EBV(+) and EBV(-) cases in Japanese population.

We have investigated 114 cases with diffuse large B-cell lymphoma (DLBCL) to clarify the characteristics of DLBCL with Epstein-Barr virus (EBV) infection. Thirteen cases (11.4%) showed EBV-encoded RNA 1 (EBER1) signals by RNA in situ hybridization. EBV-encoded latent membrane protein 1 (LMP1) and EBV-encoded nuclear antigen 2 (EBNA2) were expressed in 11 and 4 cases, respectively. Expression of CD30, Bcl-6 and immunoglobulin (Ig) was found in 92%, 31% and 23% with EBV(+) DLBCL, and in 15%, 79% and 82% with EBV(-) DLBCL, respectively. The sequence of rearranged Ig heavy chain (IgH) variable (V) region gene was analyzed in 5 cases with EBV(+) DLBCL and 61 cases with EBV(-) DLBCL. Somatic mutation was found in all cases except one with EBV(-) DLBCL. Average mutation frequency was 9.6% in EBV(+) DLBCL vs. 11.5% in EBV(-) DLBCL. The rates of replacement mutation vs. silent mutation (R / S values) in complementarity determining region II and framework region III were 2.7 and 1.5 in EBV(+) DLBCL, 2.6 and 1.4 in EBV(-) DLBCL. Crippling mutation generating a stop codon was found in 2 of 5 cases (40%) with EBV(+) DLBCL, but none of 61 cases (0%) with EBV(-) DLBCL. These findings suggest that EBV(+) DLBCL and EBV(-) DLBCL were both derived from germinal center (GC) or post-GC B cells, and EBV(+) DLBCL frequently have a non-functional IgH gene owing to crippling mutation.