EBV RNA Research Articles

Ocrelizumab B cell depletion has no effect on HERV RNA expression in PBMC in MS patients

BackgroundEpstein barr virus (EBV) infection of B cells is now understood to be one of the triggering events for the development of Multiple Sclerosis (MS), a progressive immune-mediated disease of the central nervous system. EBV infection is also linked to expression of human endogenous retroviruses (HERVs) of the HERV-W group, a further risk factor for the development of MS. Ocrelizumab is a high-potency disease-modifying treatment (DMT) for MS, which depletes B cells by targeting CD20. ObjectivesWe studied the effects of ocrelizumab on gene expression in peripheral blood mononuclear cells (PBMC) from paired samples from 20 patients taken prior to and 6 months after beginning ocrelizumab therapy. We hypothesised that EBV and HERV-W loads would be lower in post-treatment samples. MethodsSamples were collected in Paxgene tubes, subject to RNA extraction and Illumina paired end short read mRNA sequencing with mapping of sequence reads to the human genome using Salmon and differential gene expression compared with DeSeq2. Mapping was also performed separately to the HERV-D database of HERV sequences and the EBV reference sequence. ResultsPatient samples were more strongly clustered by individual rather than disease type (relapsing/remitting or primary progressive), treatment (pre and post), age, or sex. Fourteen genes, all clearly linked to B cell function were significantly down regulated in the post treatment samples. Interestingly only one pre-treatment sample had detectable EBV RNA and there were no significant differences in HERV expression (of any group) between pre- and post-treatment samples. ConclusionsWhile EBV and HERV expression are clearly linked to triggering MS pathogenesis, it does not appear that high level expression of these viruses is a part of the ongoing disease process or that changes in virus load are associated with ocrelizumab treatment.

Differentiated Papillary NUT Carcinoma: An Unexpected, Deceptively Bland Presentation of a Sinonasal Carcinoma

BackgroundIn recent years, the list of tumor entities in the sinonasal tract has significantly expanded, requiring advanced diagnostic testing. We report the case of a 32-year-old patient with an unusual NUT carcinoma originating in the maxillary sinus, which showed extensive well-differentiated, papillary squamous morphology, similar to the spectrum of the recently described DEK::AFF2 fusion-associated carcinoma.MethodsWe performed immunohistochemical and molecular studies including EBV- and HPV-testing, as well as DNA/RNA next generation sequencing.ResultsThe tumor showed predominantly exophytic papillary growth with mature squamous differentiation. An additional component harbored atypical, less differentiated basaloid tumor cells with infiltration of the adjacent stroma. Conspicuous inflammation was evident. There was no evidence of HPV DNA or EBV RNA. Next-generation sequencing revealed a NUT::NSD3 gene fusion corresponding to (“speckled-type”) immunopositivity of NUT in the tumor cells.ConclusionsWe describe a NUT::NSD3 gene fusion-associated NUT carcinoma of the sinonasal tract with a deceptively well-differentiated papillary growth pattern, thus expanding the morphological spectrum of this typically poorly differentiated neoplasm.

Lymphoepithelial carcinoma of the oral mucosa. A rare histopathologic subtype of squamous cell carcinoma

<h3>Objectives</h3> Lymphoepithelial carcinoma (LEC) represents a rare histopathologic subtype of squamous cell carcinoma (SCC), which shares morphologic similarities with non-keratinizing undifferentiated nasopharyngeal carcinoma (NPC). LEC may be etiologically associated with EBV, especially in endemic regions. Although being described in several locations of the head and neck, the oral mucosa seems to be an infrequent site of involvement. Herein, we present a case of an EBV-negative oral LEC and discuss the diagnostic challenges. <h3>Findings</h3> A 51 year-old male Filipino patient suffering from chronic kidney disease presented for the evaluation of a swelling in the right mandible, first noticed by the patient a few days ago. Clinically, the right posterior mandibular gingiva appeared enlarged and hemorrhagic, covered by ulcerated mucosa, while the adjacent teeth displayed high mobility. Extraorally, a painful and slightly movable lymph node in the right submandibular area was palpated. Radiographic evaluation revealed extensive bone loss in the right posterior mandible. With a provisional diagnosis of lymphoma or SCC, an incisional biopsy was performed. Histopathologic examination revealed invasion of the underlying connective tissue by pleomorphic cells of epithelioid morphology, occasionally arranged in syncytial islands, arising from a partially ulcerated, non-keratinized squamous epithelium. A prominent diffuse lymphocytic cell infiltrate surrounding the tumor cells was also detected. The neoplastic cells expressed pancytokeratin, CK5/6, p63, and p40, while p16, LMP-1, and in situ hybridization for EBV RNA were negative. A final diagnosis of moderately to poorly differentiated SCC with features indicative of LEC was rendered and the patient was referred for further management. <h3>Conclusions</h3> LEC should be considered in the differential diagnosis of undifferentiated malignant neoplasms displaying close association with diffuse lymphoid aggregates. The evaluation of nasopharynx is justified to rule out metastatic NPC, as the separation of these two entities microscopically is not feasible.

Clonally-Related Composite Classic Hodgkin Lymphoma and Follicular Lymphoma

Abstract Introduction/Objective Composite classic Hodgkin lymphoma and follicular lymphoma (CHLFL), defined as CHL and FL occurring simultaneously at the same site, is rare and poorly understood. While both Hodgkin/Reed-Sternberg (HRS) cells and FL are thought to be derived from germinal center B-cells, the relationship between CHL and FL when coexistent is unclear. Here, we present two cases of CHLFL and show that the CHL and FL components have a clonal relationship by FISH. Methods/Case Report Case #1 is a 50-year-old man with abdominal and mediastinal lymphadenopathy. An excised mesenteric lymph node showed two distinct components diagnostic for FL, grade 1-2 and CHL. Case #2 is a 63-year- old woman with a history of FL with transformation to diffuse large B-cell lymphoma. Cytogenetic studies showed a complex karyotype with an add(9p), del(10q), and trisomy 16. Post-treatment imaging revealed left axillary adenopathy. An excised axillary lymph node showed CHL with peripheral areas of FL, grade 3A. Both cases had areas of typical FL with BCL2-positive phenotype and no significant CD30/CD15 expression. HRS cells were CD45/CD20-negative, expressed CD30 (strong), CD15, and PAX5, and were present in a mixed inflammatory background. No EBV RNA was present by in situ hybridization. Interestingly, HRS cells in case #1 expressed both BCL6 and BCL2. FISH was performed in both cases. Case #1 had a BCL2 rearrangement in 48% of FL nuclei and in 100% of HRS cells. In case #2, targeted probes were used based on prior cytogenetic results. Here, 47% of FL nuclei and 44% of HRS cells had a 16p duplication; additionally, 32% of HRS cells had an unbalanced IGH rearrangement with loss of the IGH variable region, suggesting possible clonal evolution. No rearrangement of BCL2 or BCL6 was present. An additional 27 CHLFL cases from the literature were reviewed. CHLFL was mostly nodal and occurred in late adulthood in patients with or without a history of FL. It presented at advanced clinical stage, with a 5-year overall survival of 22%. BCL2 expression in HRS cells was common. Bone marrow involvement was 45% (5/11) and consisted of FL exclusively. Five of six tested cases demonstrated BCL2/IGH rearrangement in both FL and HRS cells. Results (if a Case Study enter NA) NA Conclusion Composite CHL and FL are often clonally related and may share a common progenitor B-cell origin – likely a germinal center B-cell – from which additional genetic abnormalities are acquired to develop two distinct lymphomas.

Abstract 3787: Reduced expression of INPP4B by promoter hypermethylation in Epstein-Barr virus-associated gastric carcinoma

Abstract Background: Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is attracting increasing attention due to distinct molecular and clinical characteristics. However, its carcinogenesis still remains unclear. Previous studies demonstrated that aberrant DNA methylation is one of the pathogenic mechanisms of EBVaGC. Inositol polyphosphate 4-phosphatase type II (INPP4B), as a component of PI3K/AKT pathway, is implicated in the regulation of crucial cellular processes and tumorigenesis. This study aims to reveal the correlation between INPP4B expression and EBV infection, as well as investigate the inactivation mechanism of INPP4B in EBVaGC. Methods: EBV status and INPP4B expression were examined in gastric carcinoma (GC) tissue microarray (301 cases) using EBV RNA probe and immunohistochemistry, respectively. The correlation of EBV status and INPP4B expression was analyzed in GC by Chi-Squared Test. The INPP4B promoter methylation was investigated in GC cell lines: SNU-719 (EBV positive), AGS (EBV negative) and SGC-7901 (EBV negative) with methylation-specific polymerase chain reaction (MSPCR). After treatment with 5-Aza-2′-deoxycytidine, the INPP4B promoter methylation and expression were detected using MSPCR, western blot, and qPCR. Results: INPP4B expression is decreased in 146 of 301 GC cases (48.5%). In 272 evaluable GC cases, EBV was present in 40 cases (14.71%). In addition, Lower expression of INPP4B is significantly correlated with EBV infection (p&amp;lt;0.05). There was a significantly decreased expression of INPP4B in EBV positive cell line (SNU-719) compared to EBV negative cell line (AGS and SGC-7901). Moreover, aberrant CpG island hypermethylation of INPP4B was found in SNU-719 rather than AGS and SGC-7901. After treatment with 5-Aza-2′-deoxycytidine, INPP4B protein and mRNA are dramatically up-regulated, while unmethylated DNA became detectable in SNU-719. Conclusions: Our results suggest that the reduction of INPP4B expression is commonly found in GC, particularly in EBVaGC. Promoter hypermethylation may be one of the mechanisms of INPP4B inactivation. Citation Format: Yiwen Ma, Jichao Tan, Weihong Dai, Yu Zhang, Yu Zhang, Rongjin Lin, Yan Xu. Reduced expression of INPP4B by promoter hypermethylation in Epstein-Barr virus-associated gastric carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3787.

Prevalence and Prognostic Impact of CD30 and EBV Co-Expression in Korean Patients with EBV-Positive DLBCL of the Elderly

The prognostic impact of EBV-positivity in EBV-positive DLBCL of the elderly is controversial. Previous Asian studies have shown consistent poor prognostic impact of EBV-positivity. Recent Western study has shown no significant prognostic impact of EBV-positivity alone. However, the study found that CD30 and EBV co-expression is associated with an adverse outcome and suggested CD30 co-expression might be associated with different prognostic impact of EBV-positivity between Asian and Western populations.We retrospectively analyzed 26 Korean patients with EBV-positive DLBCL of the elderly diagnosed at Samsung Medical Center and had adequate amount and quality of paraffin-embedded biopsy specimen for additional immunostaining of CD30. EBV RNA was detected by an ISH technique and a positive reaction was defined as more than 20% of examined cells showing nuclear positivity. Cell of origin was assigned using Hans immunohistochemical algorithm.Prevalence of CD30 co-expression in EBV-positive DLBCL of the elderly was 65.4% (17/26) and 50.0% (13/26) using a >0% cut-off and a >30% cut-off, respectively. There was no statistically significant difference of survival between cases with CD30 co-expression and case without CD30 co-expression. However, the cases with CD30 co-expression showed a modest trend of worse overall survival (OS) (median OS, 12.5 months [95%CI 1.5-23.4] versus not reached, P = 0.279) and progression-free survival (PFS) (median PFS, 7.4 [95%CI 5.7-9.1] versus 34.2 months [95%CI 0.0-89.1], P = 0.430) using >0% cut-off. Among 17 patients with CD30 and EBV co-expression, most of patients (88%, 15/17) showed non-germinal center B-cell like (GCB) DLBCL.Substantial proportion of Korean patients with EBV-positive DLBCL of the elderly showed CD30 co-expression. CD30 co-expression in EBV-positive DLBCL of the elderly showed a modest trend of an adverse outcomes without statistical significance. Further validation study with larger population is warranted. Considering previously known contradictory trend of better outcome in CD30 positive cases with EBV-negative and GCB type DLBCL, we also need to investigate the possible distinct pathobiology of CD30 positivity in EBV-positive DLBCL of the elderly. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

CD30 Expression in Diffuse Large B-Cell Lymphoma

AbstractAbstract 1558▪▪This icon denotes a clinically relevant abstract Background:Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease with a variable clinical course. The addition of rituximab (R) to CHOP combination chemotherapy has improved overall (OS) and progression free survival (PFS) but some patients progress despite treatment and alternative therapies are needed. Brentuximab vedotin is an antibody-drug conjugate that targets CD30. It is efficacious in the treatment of relapsed Hodgkin lymphoma and relapsed systemic anaplastic large cell lymphoma, two lymphomas associated with CD30 expression. The association between DLBCL and CD30 expression has not been well described. The aim of this study was to examine CD30 expression in DLBCL. Design:395 cases of formalin-fixed paraffin-embedded DLBCL (excluding PMBCL) in a tissue microarray were independently evaluated by two pathologists for expression of CD30, CD10, BCL6, and MUM1 by immunohistochemistry (IHC) and EBV RNA (EBER) by in situ hybridization. CD30 expression was correlated with cell of origin (COO) phenotype, OS and PFS, EBV infection, International Prognostic Index (IPI) score and CD30 mRNA expression. The COO phenotype, germinal center B-cell like (GCB) or non-GCB, was determined by IHC using the Hans algorithm. CD30 mRNA expression and COO genotype by gene expression profiling (GEP) were determined using Affymetrix U133 2.0 Plus arrays (n=170). Outcome analysis only included patients treated with R-CHOP chemotherapy. CD30 was considered positive by IHC if any malignant cells exhibited membranous staining. The threshold for calling higher CD30 expression by GEP was determined using X-Tile software. Results:25% (95/385) of DLBCL cases expressed CD30 by IHC with excellent concordance between two observers (r = 0.94). CD30 expression trended towards a non-GCB phenotype but was not significantly different (p=0.067). CD30 expression was not associated with PFS or OS in all R-CHOP treated cases (n=313); however, it was associated with a prolonged PFS in GCB-DLBCL (n=147) (p=0.019). In GCB-DLBCL CD30 expression remained an independent predictor of PFS in a multivariate analysis with IPI (p=0.038). CD30 expression by IHC was significantly associated with higher levels of CD30 mRNA (p=0.002). ABC-DLBCL exhibited significantly higher expression levels of CD30 mRNA (p=0.037). Higher CD30 mRNA expression was associated with a prolonged PFS in all R-CHOP treated DLBCL (p=0.012) as well as in DLBCL with a GCB-genotype (p=0.008), but not ABC or U-genotypes. Higher CD30 mRNA expression was also associated with a prolonged OS in the GCB-genotype (p=0.022). In the GCB-genotype higher CD30 mRNA expression remained an independent predictor of PFS, but not OS, in a multivariate analysis with IPI (p=0.037). EBV was identified in 3% of DLBCL (11/391), all of which exhibited a non-GCB phenotype (p=0.001) and were almost exclusively positive for CD30 expression (10/11)(p=<0.001). Conclusions:CD30 is expressed in approximately 25% of DLBCL and brentuximab vedotin could be considered for study in combination with traditional front-line therapies or as an alternative therapy in the relapsed or refractory disease. CD30 immunohistochemistry may be useful as a prognostic marker in R-CHOP treated GCB-DLBCL and the significant association of CD30 with EBV-positive non-GCB DLBCL suggests a distinct pathobiology for these cases. Disclosures:Slack:Seattle Genetics: Research Funding. Steidl:Seattle Genetics: Research Funding. Gascoyne:Seattle Genetics: Research Funding.

H. pylori Negative Diffuse Russell Body Gastropathy

Purpose: A 71-year-old male with a history of atrial fibrillation on warfarin and a normal colonoscopy 5 years ago presented with melena and a hemoglobin of 4.1 g/dL. He denied alcohol or NSAID use. No personal or family history of GI disease. EGD revealed diffuse finely nodular gastric mucosa (Figure 1). Biopsies were significant for plasma cells containing numerous round eosinophilic globules positive for CD 138 consistent with Russell bodies (Figure 2). CKMAK-6 and CKAE1+3 were negative confirming these were plasma cells and not epithelial in origin. The plasma cells were κ- and λ-immunostain positive supporting their polyclonal nature. There was no evidence of malignancy. Small bowel capsule endoscopy and abdominal CT scan were negative. Russell bodies are eosinophillic cytoplasmic inclusions within plasma cells resulting from the accumulation of immunoglobulins within the endoplasmic reticulum. Russell body gastritis is rare and was first reported in 1998. Russell bodies have been associated with H. pylori gastritis (in 7 of 10 reported cases), HIV gastritis, EBV, Barrett's esophagitis, duodenitis, histoplasmosis, TB, multiple myeloma, endocervical polyps, peridontitis, and actinic keratosis. There have been recent reported cases of Mott cell (a plasma cell containing Russell bodies) proliferation in gastric carcinoma encoded with EBV RNA, signet ring cell gastric cancer, and MALToma. The risk of malignancy with Russell bodies is unknown as there is no long term follow-up. This patient was unique in that he had diffuse nodularity with Russell bodies, and was H. pylori negative and without associated aspirin, NSAID, alcohol use, infection, or malignancy.Figure 1: No Caption available.Figure 2: No Caption available.

Oral dysplasia and squamous cell carcinoma: Correlation between increased expression of CD21, Epstein-Barr virus and CK19

Epstein-Barr virus is an orally transmitted human gammaherpesvirus that infects B lymphocytes and epithelial cells. Although most primary infections are asymptomatic, long term carriage of the virus can be associated with either lymphoid or epithelial malignancies. The association of EBV with oral squamous cell carcinomas is sporadic and it is uncertain if the virus is involved in initiation of the tumor or, possibly, in its progression. Complement receptor type 2, CR2 or CD21, is a receptor for the major attachment protein of EBV, which significantly enhances epithelial cell infection, but its expression on normal tissues is restricted to tonsil and adenoid epithelium. As cells become dysplastic they are reported to express higher levels of CK19. We sought to evaluate whether CD21 and CK19 expression change as oral epithelial cells outside Waldeyer's ring become dysplastic. Epithelial cells were isolated by laser capture microdissection and levels of CD21, CK19 and EBV RNA were measured by quantitative reverse transcriptase PCR. We report that expression of CD21 increases in frequency and intensity as oral epithelial cells become more dysplastic and that expression correlates with an increase in infection by EBV. Tumors or dysplastic lesions that carry EBV also generally express higher levels of CK19 than those that do not. The findings suggest that dysplasia may make cells more susceptible to infection by EBV and that infection by the virus may alter the phenotype of the infected cell in a manner which could affect prognosis.

An Uncommon Cause of Refractory Ascites in an Immunocompromised Patient

Purpose: Primary Effusion Lymphoma (PEL) is a rare form of Non-Hodgkin's lymphoma that involves serous body cavities (pleural, pericardial, or peritoneal) with lymphomatous effusions in the absence of lymphadenopathy or organomegaly. PEL is associated with HIV, HHV-8, EBV(formally designated as Human Herpes Virus 4), Hepatitis B or Hepatitis C. PEL is one of the rarest of the AIDS-related lymphomas, accounting for less than 1-4 percent of cases. PEL often presents with rapidly progressive effusions and generally has a poor prognosis. By flow cytometry, the antigens expressed in PEL are CD38, CD71, HLA-DR, CD30, and CD45. B-cell markers are usually negative. Cytomorphologically, all cases show atypical to anaplastic morphology. Median survival after diagnosis without treatment is approximately 2 to 3 months. Even with aggressive chemotherapy, median survival is extended to only 6 months. Methods: We describe a case of a 28-year-old African American male patient with a past medical history of AIDS and an absolute CD 4 T lymphocyte count of 35 cells/ml and non-compliance with Highly Active Anti Retroviral Therapy (HAART) presenting with abdominal pain, fever and shortness of breath. Patient was found to have ascites and pleural effusion on admission. The analysis of the ascitic and pleural fluid showed the effusion was most likely malignant. An examination of the fluid by cytology showed large atypical lymphocytes with abundant basophilic cytoplasm and irregular nuclei containing multiple prominent nucleoli. Flow cytometry of the effusion turned out to be a B cell lymphoma. The neoplastic cells showed strong positivity for HLA DR, CD 20, CD 30, CD 38, CD 45, EBV RNA (on in situ hybridization), cytoplasmic lambda light chain and focal positivity for HHV 8 Latency Associated Nuclear Antigen (LANA). He had recurrent episodes of ascites, pleural and pericardial effusion despite repeated pleural and ascitic fluid drainage. The patient expired on day 42, before initiation of chemotherapy, because of hypoxic respiratory failure. Conclusion: It is essential for clinicians and pathologists to have a high index of suspicion of PEL in HIV positive patients who have effusions without solid tumors or lymphadenopathy. Basic immunohistochemistry is essential for definitive diagnosis.

Characterization and treatment of chronic active Epstein-Barr virus disease: a 28-year experience in the United States

AbstractChronic active EBV disease (CAEBV) is a lymphoproliferative disorder characterized by markedly elevated levels of antibody to EBV or EBV DNA in the blood and EBV RNA or protein in lymphocytes in tissues. We present our experience with CAEBV during the last 28 years, including the first 8 cases treated with hematopoietic stem cell transplantation in the United States. Most cases of CAEBV have been reported from Japan. Unlike CAEBV in Japan, where EBV is nearly always found in T or natural killer (NK) cells in tissues, EBV was usually detected in B cells in tissues from our patients. Most patients presented with lymphadenopathy and splenomegaly; fever, hepatitis, and pancytopenia were common. Most patients died of infection or progressive lymphoproliferation. Unlike cases reported from Japan, our patients often showed a progressive loss of B cells and hypogammaglobulinemia. Although patients with CAEBV from Japan have normal or increased numbers of NK cells, many of our patients had reduced NK-cell numbers. Although immunosuppressive agents, rituximab, autologous cytotoxic T cells, or cytotoxic chemotherapy often resulted in short-term remissions, they were not curative. Hematopoietic stem cell transplantation was often curative for CAEBV, even in patients with active lymphoproliferative disease that was unresponsive to chemotherapy. These studies are registered at http://www.clinicaltrials.gov as NCT00032513 for CAEBV, NCT00062868 and NCT00058812 for EBV-specific T-cell studies, and NCT00578539 for the hematopoietic stem cell transplantation protocol.

Primary effusion lymphoma associated with Human Herpes Virus-8 and Epstein Barr virus in an HIV-infected woman from Kampala, Uganda: a case report

IntroductionPrimary effusion lymphoma is a recently recognized entity of AIDS related non-Hodgkin lymphomas. Despite Africa being greatly affected by the HIV/AIDS pandemic, an extensive MEDLINE/PubMed search failed to find any report of primary effusion lymphoma in sub-Saharan Africa. To our knowledge this is the first report of primary effusion lymphoma in sub-Saharan Africa. We report the clinical, cytomorphologic and immunohistochemical findings of a patient with primary effusion lymphoma.Case presentationA 70-year-old newly diagnosed HIV-positive Ugandan African woman presented with a three-month history of cough, fever, weight loss and drenching night sweats. Three weeks prior to admission she developed right sided chest pain and difficulty in breathing. On examination she had bilateral pleural effusions.Haematoxylin and eosin stained cytologic sections of the formalin-fixed paraffin-embedded cell block made from the pleural fluid were processed in the Department of Pathology, Makerere University, College of Health Sciences, Kampala, Uganda. Immunohistochemistry was done at the Institute of Haematology and Oncology "L and A Seragnoli", Bologna University School of Medicine, Bologna, Italy, using alkaline phosphatase anti-alkaline phosphatase method. In situ hybridization was used for detection of Epstein-Barr virus.The tumor cells were CD45+, CD30+, CD38+, HHV-8 LANA-1+; but were negative for CD3-, CD20-, CD19-, and CD79a- and EBV RNA+ on in situ hybridization. CD138 and Ki-67 were not evaluable. Our patient tested HIV positive and her CD4 cell count was 127/μL.ConclusionsA definitive diagnosis of primary effusion lymphoma rests on finding a proliferation of large immunoblastic, plasmacytoid and anaplastic cells; HHV-8 in the tumor cells, an immunophenotype that is CD45+, pan B-cell marker negative and lymphocyte activated marker positive. It is essential for clinicians and pathologists to have a high index of suspicion of primary effusion lymphoma when handling HIV positive patients who have effusions without palpable tumor masses. Basic immunohistochemistry is essential for definitive diagnosis.

Clinicopathological Features of Nodular Sclerosis-Type Classical Hodgkin Lymphoma In the Elderly: Multicenter Study of Hodgkin Lymphoma In Japan

AbstractAbstract 2677 Background:Classical Hodgkin lymphoma (CHL), which is characterized by the presence of Hodgkin and Reed Sternberg (H-RS) cells in a background of non-neoplastic inflammatory cells, is divided into four histological subgroups, nodular sclerosis (NSCHL), mixed cellularity (MCCHL), lymphocyte-rich, and lymphocyte depletion. While NSCHL in young adults is characterized by a mediastinal mass and good prognosis, the clinicopathological characteristics of NSCHL in the elderly (NSCHL-e) remain uncertain. Patients and methods:Enrolled patients were diagnosed with CHL between 1986 and 2006 as part of the Hodgkin Lymphoma's Multicenter Study Group. To better characterize NSCHL-e, we compared the clinicopathological profiles of 84 NSCHL-e patients aged 50 or over with 237 NSCHL-y patients aged 49 or younger and 302 with MCCHL. Results:The total of 743 CHL patients consisted of 496 men and 247 women with a median age of 48 years (range, 15– 89 years). The pathological diagnoses were NSCHL in 324 patients (43%) and MCCHL in 303 (41%). NSCHL patients showed a bimodal age distribution, with an initial peak in their 20s and a second small peak in their 60s. We categorized the former as NSCHL-y (49 or younger) and the latter as NSCHL-e (50 and over). NSCHL-e patients were characterized by male predominance and a more advanced clinical stage (53%) than NSCHL-y. Immunophenotypically, H-RS cells had the prototypic immunophenotype of CD15+ CD30+ and Pax5+. NSCHL-e cases showed a significantly higher rate of CD20 (24%) than NSCHL-y (8%, P = 0.001). Furthermore, H-RS cells in 29 of 75 (39%) patients with NSCHL-e were positive for EBV RNA transcripts by in situ hybridization, whereas only 7% of NSCHL-y cases were EBER-positive (P < 0.0001) (Table). Regarding NSCHL-e and MCCHL, no significant difference between these patients was seen in clinical characteristics. Immunophenotypically, NSCHL-e patients showed significantly higher rates for CD3 and TIA-1, while MCCHL patients showed higher EBV positivity (75%). Fifty-five of 63 patients received systemic multi-agent chemotherapy as first-line treatment, consisting of doxorubicin, bleomycin, vinblastine, and dacarbacin (ABVD) in 38 patients; CHOP in 8; C-MOPP in 8; and BEACOPP in 1. Overall, 51 patients responded to first-line treatment, 39 with complete response and 12 with partial response. Disease-specific survival of NSCHL-e was poorer than that of NSCHL-y (P < 0.001) but similar to that of MCCHL (P = 0.43) (Figure). Conclusion:NSCHL-e is characterized by an unfavorable prognosis and different clinicopathological features to NSCHL-y, which is considered as typical NSCHL. A number of cases of NSCHL-e might have been associated with MCCHL, with most being EBV-positive. These results suggest the limitations of current histological subgroupings for CHL. [Display omitted] Disclosures:Matsushita:Pfizer CO.: Membership on an entity's Board of Directors or advisory committees, Research Funding; Baxter Co.: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Clinical features and treatment outcomes of angioimmunoblastic T-cell lymphoma.

8048 Background: Angioimmunoblastic T-cell lymphoma (AITL) accounts for up to 15-20% of all peripheral T-cell lymphomas. Typically, the disease follows an aggressive course with systemic symptoms and an overall poor prognosis. The primary objectives of this analysis were 1) to assess the association between complete response status after first-line chemotherapy and the other known prognostic factors; 2) to estimate the overall survival (OS) and progression-free survival (PFS) by patients' prognostic factors. Methods: Sixty-three patients diagnosed with AITL between 1996 and 2009 were included in the study. Patients characteristics were: 57% male, median age 65 (range: 35- 83), 94% stage III-IV, 46% IPI of ≥3. Eighty percent of the patients presented with B symptoms and 18% of them had a history of previous chemical exposure. Most of the patients were initially treated with anthracycline-based chemotherapy (87%) and 11 (17%) patients received rituximab in combination with chemotherapy. Results: Overall response rate to initial chemotherapy was 78% with 65% of patients achieving a CR. The only factor associated with achievement of CR was low IPI (p= 0.05). With a median follow-up of 20 months (range: 2-152), the 5-year and 10-year OS rates of all patients were 47% and 23% respectively. The 5-year and 10-year PFS rates were 18% and 15% respectively. Performance status and IPI were both predictive prognostic factors for survival (p=0.005 and 0.04 respectively) whereas only IPI was predictive for PFS (p=0.03). The presence of EBV RNA detected by in-situ hybridization was associated with a better PFS (p=0.02). Conclusions: This study confirms the poor prognosis of patients with AITL when treated with standard anthracycline-based chemotherapy regimens. IPI remains the strongest predictive factor for both OS and PFS. Novel therapies are needed to improve the outcome of this disease. No significant financial relationships to disclose.

Cytotoxic Molecule (CM)-Positive Classical Hodgkin Lymphoma: a Clinicopathologic Study in Comparison with Nodal Peripheral T-Cell Lymphoma of Not Otherwise Specified Type Possessing CM Expression.

Abstract 1549Poster Board I-572Classical Hodgkin lymphoma (CHL) is characterized by Hodgkin and Reed Sternberg (H-RS) cells, which are B-cell origin in many cases. Recently, we highlighted an adverse prognostic significance of cytotoxic molecule (CM) expression among CHL patients (Asano N, et al. J Clin Oncol. 2006). However, the clinical characteristics of CM-positive CHL still remain controversial. We here document the clinicopathologic profiles of 35 patients with CM-positive CHL, consisting of 23 men and 12 women with a median age of 50 years (range, 16 - 84 years). All patients had lymphoadenopathy, and 14 cases showed mediastinal involvement at presentation. Physical findings included hepatomegaly and splenomegaly in six and five patients, respectively. Four patients had a bulky mass, and nine showed stage IV disease. As for laboratory data, four patients had elevated white blood cell counts (greater than 15.0 × 103/mm3). Anemia (hemoglobin level; less than 10.5 g/dl) was also present in six patients. The pathological diagnoses were nodular sclerosis type (NS) in 22 cases and mixed cellularity (MC) in 12 cases. The H-RS cells of CM-positive CHL had a prototypic immunophenotype of CD15+ CD30+ and Fascin+. All of the cases completely lacked CD20 positivity on H-RS cells, while the expression of CD3e and CD45RO was found in 2 and 1, respectively. The H-RS cells were further positive for EBV RNA transcripts in 14 of 32 (44%) cases studied by in situ hybridization method. All except three cases were negative for Pax5. A clonal TCR-g chain gene arrangement was undetected in any of the cases with successful amplification of control GAPDH DNA by PCR analysis. No cases show B-cell clonality. 27 of 35 patients received systemic multi-agent chemotherapy consisting of first-line treatment regimens as follows: doxorubicin, bleomycin, vinblastine, and dacarbacin (ABVD) (22 patients); cyclophosphamide, doxorubicin, vincristin, and prednisone (CHOP) (2 patients); two with C-MOPP; and one BEACOPP regimens. Three patients died within 6 months, before completing induction treatment because of disease progression. Overall, 22 patients responded to first-line treatment: 13 with complete response and 9 with partial response. 13 of them had relapses, and 8 died with a clinical course ranging from 7 to 142 months. Effective therapeutic approaches should be explored for CM-positive CHL patients, who resist standard treatment for CHL. Comparing CM-positive nodal peripheral T-cell lymphomas of not otherwise specified type (PTCL-N)(n=55) with CM-positive CHL(n=35), no significant differences were detected in clinical parameters except for the frequency of elevated lactate dehydrogenase level in CM-positive PTCL-N. Immunophenotypically, CM-positive CHL cases showed significantly higher rates for CD15, CD30, and Fascin expression, while CM-positive PTCL-N cases showed significantly higher CD3e, CD8, CD45RO, and CXCR3 positivity. The survival curve of CM-positive CHL showed poorer prognosis than that of CM-negative CHL (P = .0002) and better than that of CM-positive PTCL-N (P = .002) (Figure). These findings suggest that CM-positive CHL is characterized by an unfavorable clinical feature, although their histologic and phenotypic features are more suggestive of CHL. Moreover, one case in CM-positive CHL had presented the skin lesion as a CD30-positive lymphoproliferative disorder or an ALK-negative anaplastic large cell lymphoma (ALCL) at the time of recurrence. These findings suggest that CM-positive CHL represent a distinct variant form based on clinicopathologic and phenotypic traits beyond the framework of CHL. Significant overlaps in biologic and morphologic features has been identified in CHL and non-Hodgkin lymphoma. CM-positive CHL cases may be ‘intermediate lymphoma‘ between CHL and PTCL or ALK-negative ALCL. Further studies, including array CGH profiling analysis are needed to clarify the origin of CM-positive CHL. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Angioimmunoblastic T-cell lymphoma with hyperplastic germinal centres: a neoplasia with origin in the outer zone of the germinal centre? Clinicopathological and immunohistochemical study of 10 cases with follicular T-cell markers

Angioimmunoblastic T-cell lymphoma is an aggressive peripheral T-cell lymphoma whose natural history is not fully understood. Up to 17% of cases can present histologically with hyperplastic germinal centres (pattern I). The accurate recognition of Angioimmunoblastic T-cell lymphoma with pattern I remains a challenge and therefore the aim of this study is to phenotypically and morphologically characterize this variant with the use of the follicular helper T-cell (TFH) markers PD1, CXCL-13 and ICOS. Out of the 88 Angioimmunoblastic T-cell lymphoma cases reviewed, 10 showed hyperplastic follicles. Molecular probe methods for the detection of T-cell and B-cell clonality, as well as in-situ hybridization probes for EBV RNA expression, were carried out to leave no question as to the establishment of the diagnosis in each case. Of the 10 cases, all (100%) showed strong positive PD1 staining in perifollicular areas and in neoplastic cells surrounding small veins. CXCL13 and ICOS showed a similar staining pattern. By contrast, CD10 was found to only weakly label the neoplastic T cells, with only 5–10% of the target cell population staining for this marker. EBV was found in 9/10 cases. Clinically, 8/9 cases presented with stage IIIB/IVB and in 2/10 cases consecutive biopsies showed ‘progression’ from pattern I to classical Angioimmunoblastic T-cell lymphoma. In conclusion we have shown that the TFH cells markers PD1, CXCL13 and ICOS are useful adjuncts in the diagnosis of Angioimmunoblastic T-cell lymphoma with hyperplastic germinal centres. PD1 also highlighted the presence of neoplastic cells in the outer zone of lymphoid follicles, suggesting that Angioimmunoblastic T-cell lymphoma (pattern I) may originate from TFH cells in this region, in accordance with previous immunological studies. As the majority of cases in our series presented clinically with advanced stage disease, progression from pattern I to classical Angioimmunoblastic T-cell lymphoma may represent histological evolution rather than clinical progression.

Retrospective Review of Angiommunoblastic T-Cell Lymphoma: The MD Anderson Cancer Center Experience.

Angioimmunoblastic T-cell Lymphoma (AITL) represents about 4–6% of all lymphomas, but up to 15–20% of all T-cell lymphomas. AITL usually presents in the 6th to 7th decade of life as an advanced stage, systemic disease with B-symptoms, lymphadenopathy, bone marrow involvement, rash, and autoimmune abnormalities. The disease follows an aggressive course and typically portends a poor prognosis. Nevertheless, there are no widely accepted treatment guidelines that have shown significant efficacy or improved benefit in this disease.We report our single institution experience with AITL over the past 10 years.We identified 33 cases of AITL seen at MDACC from 1996–2006. We retrospectively reviewed these cases to analyze clinicopathologic characteristics, treatment regimens, and response rates. The median age at diagnosis was 67 years (43–79), and there was a male to female ratio of 2:1. Thirty-two patients (97%) presented with stage 3 or 4 disease, 16 patients (49%) had bone marrow involvement, and 23 patients (70%) had B symptoms at presentation. Stratifying 32 patients by IPI, 12.5% were classified as low risk, 43.8% as low-intermediate, 28% as high-intermediate, and 15.6% as high risk. A skin rash was present in 16 patients (49%), while 4 patients (12%) had significant inflammatory arthralgias, and 5 patients (15%) had an autoimmune hematologic disorder. Interestingly, 6 patients (18%) had known exposure to industrial chemicals, 8 patients (24%) had a preceding or concurrent second malignancy, and 13 out of the 15 cases (87%) tested for EBV were positive for EBV RNA. Most patients were treated with an anthracycline based regimen. Of these, 6 patients were treated with an intense, alternating triple therapy regimen, 4 patients received a hyperfractionated cyclophosphamide based regimen, and 16 patients received a CHOP-like regimen. Of these 16 patients, 4 also received rituximab in combination with CHOP. Out of 25 evaluable patients after 1st line therapy, 17 patients (68%) achieved CR, 6 patients (24%) achieved PR, and 2 patients (8%) were found to be primary refractory. In those patients with CR, the median duration of 1st remission was 30.5 months (1–96), with a median follow up of 37.6 months (2–101). The probabilities of 1,2, and 3 year survival were 80%, 67%, and 57% respectively. The duration of remission by type of therapy was found to be higher in patients receiving the more intense chemotherapy regimens when compared to CHOP alone. Overall survival of patients treated with CHOP vs. non-CHOP regimens is shown in figure 1. Although the length of followup is still short, notably, only 1 out of 4 patients treated with rituximab have relapsed. The one case had skin-only relapse and has achieved a 2nd remission. The one patient who was treated with upfront high dose chemotherapy and autologous transplant continues to maintain one of the longest durations of remission in our series. Overall response rate and duration of remission in our series was slightly higher than that reported in the literature. Strategies such as early referral to transplant and addition of rituximab to CHOP-like regimens need to be investigated further, as we work to improve the outcome of this disease. [Display omitted]

Favorable Outcomes with CHOP Therapy in Plasmablastic Lymphoma in HIV Positive Patients.

Plasmablastic lymphoma (PBL) is a rapidly progressive variant of diffuse large-cell lymphoma predominantly diagnosed in HIV positive patients. It is frequently associated with latent EBV infection and is often lacking expression of CD20 and CD45. A retrospective study of HIV positive patients diagnosed with PBL to determine outcomes after CHOP therapy. Two HIV positive patients diagnosed with PBL and treated with CHOP were identified. The first patient is a 48 year old male with CD4 count of 80 cells/mm3 on HAART, who presented with a rectal mass. Pathologic exam revealed granulation tissue with marked inflammation. A second colonoscopy because of ongoing rectal bleeding was done. Histologic exam was interpreted as infiltrating poorly differentiated carcinoma. The third biopsy obtained via colonoscopy (after developing bowel obstruction) was reported as a high grade tumor with neuroendocrine features. The patient received one dose of cisplatin 60 mg/m2 and irinotecan 60 mg/m2 and radiation therapy. Patient underwent a diverting colostomy. The immunohistochemical exam of the tissue was negative for CD20, CD3 and positive for CD 138, suggestive of the diagnosis of plasmablastic lymphoma. In situ hybridization for EBV was positive. The patient received 6 cycles of CHOP (cytoxan 750 mg/m2, vincristine 1.4 mg/m2, doxorubicine 50/m2 and prednisone 100 mg) chemotherapy and 6 more cycles of radiation therapy. He is in complete remission 16 months after the diagnosis, and continues to be followed. The second patient is a 44 year old female on HAART with CD4 count of 415 cells/mm3, who presented with the swelling of the right jaw for three weeks. The histologic exam of the tissue, obtained by the incision biopsy of the mass, favored plasmablastic lymphoma. Immunohistochemical analysis tested positive for CD3, CD138 and CD45. In situ hybridization for EBV RNA was positive. Patient received 3 cycles of CHOP regimen and radiation to the jaw. She has not shown any evidence of relapse and is in complete remission 22 months after diagnosis, and continues to be followed. Durable remissions, 16 and 22 months, were achieved with CHOP therapy in HIV positive patients with PBL. This study supports the use of CHOP as an effective treatment of PBL in HIV positive patients.

Chronic active Epstein-Barr virus infection associated with mutations in perforin that impair its maturation

AbstractChronic active Epstein-Barr virus infection (CAEBV) is a rare disease in which previously healthy persons develop severe, life-threatening illness. Mutations in the perforin gene have been found in familial hemophagocytic lymphohistiocytosis, which shares some features with CAEBV. We studied a patient who died at age 18, 10 years after the onset of CAEBV. The patient had high titers of antibodies to EBV, EBV RNA in lymph nodes, T-cell lymphoproliferative disease, and hemophagocytic lymphohistiocytosis. DNA sequencing showed novel mutations in both alleles of the perforin gene that resulted in amino acid changes in the protein. The quantity of the native form of perforin from the patient’s stimulated peripheral blood mononuclear cells (PBMCs) was extremely low and immunoblotting showed accumulation of an uncleaved precursor form of perforin. Stimulated PBMCs from the patient were defective for Fas-independent cytotoxicity. These data imply that mutations in this patient resulted in reduced perforin-mediated cytotoxicity by his lymphocytes. This is the first case in which perforin mutations have been shown to result in accumulation of the uncleaved, immature form of perforin. Mutations in the perforin gene are associated with some cases of CAEBV with hemophagocytic lymphohistiocytosis.

Isolated pleural PTLD after cardiac transplantation

Preamble: Epstein–Barr virus infection (EBV) and immunosuppression promote emergence of posttransplant lymphoproliferative disorders (PTLD) in patients undergoing organ transplantation. Objective: We report a case of PTLD confined to the pleura. Findings: The patient was a 62-year-old male who had undergone cardiac transplant in 1993 for ischemic heart disease. Seven years later, he presented with dyspnea and bilateral pleural effusions. The CT scan revealed left sided pleural base thickening. The cytology of the pleural fluid and fine needle aspirate of the pleura was both suggestive of PTLD. However, the tissue submitted for ancillary studies did not contain the diagnostic material. A clinical decision was made to withdraw immunosuppressive therapy and start rituximab. His clinical course was complicated by Pneumocystis carinii pneumonia and he died 4 months after the diagnosis of PTLD. Autopsy revealed bilateral pleural effusions with pleural nodules involving the visceral and parietal pleura of both lungs. Immunohistochemistry demonstrated B cell lineage with κ/λ ratio of 1. PCR studies done on the pleural nodules (postmortem specimen) revealed the presence of EBV DNA and absence of human herpes virus 8 (HHV8) DNA. In situ hybridization revealed positive staining for EBV RNA within the neoplasm. Conclusion: Pleural-based PTLD is rare. Cytology in conjunction with immunophenotyping and molecular studies can be useful for a definitive diagnosis. In our case, cytology sample was suggestive of PTLD. PCR studies performed on the antemortem specimen confirmed the presence of monoclonal IgH gene rearrangement, while the postmortem specimen revealed oligoclonal IgH gene rearrangement. The change from monoclonal to oligoclonal IgH gene rearrangement suggests reversion of monoclonal to polyclonal PTLD following rituximab and CHOP therapy. We also demonstrated EBV DNA and RNA in the tumor nodules, supporting EBV-induced PTLD.