EBER Gene Expression in Epstein-Barr Virus-Associated Hematopoietic Neoplasms

Epstein-Barr virus encoded RNA's (EBER) are small RNA species found in cells latently infected by the virus. The physiological function of these molecules is currently a matter of speculation. Nonetheless, their presence in extremely high copy number has made it possible to reliably detect the Epstein-Barr virus by in-situ hybridization, in human tissues routinely fixed with formalin and embedded in paraffin. Such studies have enhanced our understanding of a number of hematologic malignancies, particularly Hodgkin's disease, angiocentric im-munoproliferative lesions and angio-immunoblastic lymphadenopathy. In addition, sequential EBER in-situ hybridization studies on lymphoid tissues should enable oncologists to monitor the development of lymphoproliferative disorders occurring in the setting of organ transplantation, AIDS and hereditary immunodeficiencies.     

Epstein-Barr virus in cutaneous lymphomatoid granulomatosis

Epstein-Barr virus (EBV) has been associated with various extra-cutaneous lymphoproliferative diseases and it has been suggested that EBV may have a similar aetiological role in the skin. In this study, 10 biopsies from 7 cases of primary cutaneous lymphomatoid granulomatosis have been analyzed, using in situ hybridisation, for the presence of EBV encoded RNAs (EBER-1 and EBER-2). Only one case showed positive staining with the EBER probes and it is concluded that, in the skin, the relationship between EBV and lymphomatoid granulomatosis is variable. The role of EBV as an aetiological agent in primary cutaneous lymphomatoid granulomatosis appears less important than in primary respiratory disease.     

Plasma EBV-DNA monitoring in Epstein-Barr virus-positive Hodgkin lymphoma patients

Epstein-Barr virus (EBV) is associated with approximately one-third of Hodgkin lymphoma (HL) cases. EBV-DNA is often present in the plasma and whole blood of EBV-associated HL patients. However, the significance of EBV-DNA monitoring is debated. In a cohort of 165 adult HL patients, EBV-DNA viral load was prospectively monitored both in the plasma and whole blood. Diagnostic tissue samples of all patients were histologically reviewed; in 72% nodular sclerosis was detected, 24% presented with mixed cellularity (MC), and 5% had other type of HL. Tissues from 150 patients were also analyzed for the presence of latent EBV infection using in situ hybridization for EBV-encoded RNA (EBER) and immunohistochemistry for latent membrane protein (LMP1). Using these methods, 29 (19%) patients were classified as EBV positive. Using real-time quantitative PCR, 22 (76%) of EBV-positive HL patients had detectable EBV-DNA in the plasma and 19 (66%) patients in whole blood prior to therapy. In the group of EBV-negative HL cases, three (2%) patients had detectable plasma EBV-DNA and 30 (25%) patients whole blood EBV-DNA before treatment. EBV-positive HL was significantly associated with EBV-DNA positivity both in the plasma and whole blood in pretreatment samples, increasing age and MC subtype. Serial analysis of plasma EBV-DNA showed that response to therapy was associated with decline in viral load. Moreover, significantly increased plasma EBV-DNA level recurred before disease relapse in one patient. Our results further suggest that the assessment of plasma EBV-DNA viral load might be of value for estimation of prognosis and follow-up of patients with EBV-positive HL.     

Primary cutaneous CD56 positive lymphoma: a diagnostic conundrum in an unusual case of lymphoma

We present an unusual case of a CD56-positive T-cell lymphoma exhibiting immunophenotypic characteristics of both γδ T-cell lymphoma and extranodal NK/T-cell lymphoma, nasal-type. The patient presented with a 2-month history of rapidly progressive, pruritic and cutaneous nodules on his arms. A biopsy showed a dense pan-dermal infiltrate of markedly atypical CD3-positive lymphocytes, compatible with tumor stage cutaneous T-cell lymphoma. Retrospective review of a preceding biopsy and flow cytometric analysis, performed at an outside institution, showed strong expression of surface CD3, CD7, CD43 and γδ T-cell receptor (TCR), findings consistent with a diagnosis of cutaneous γδ T-cell lymphoma. In light of these data, we performed additional studies that showed diffuse positive staining of the atypical lymphocytes for CD56, CD4 and CD43 as well as Epstein-Barr virus-encoded small nonpolyadenylated RNA (EBER). Interestingly, this case displays characteristic features of γδ T-cell lymphoma, with strong surface expression of CD3 and γδ-TCR, as well as characteristics of natural killer (NK)/T-cell lymphoma, including expression of CD4 and EBER positivity, that represent two separate categories in the current classification of cutaneous lymphomas. Taken together, these findings underscore the difficulty of rendering an unambiguous classification of the presented neoplasm given the close ontogenetic relationship between NK and cytotoxic T-cells and highlight the need for continued reevaluation of the current classification system.     

Cytopathologic features and differential diagnostic considerations of primary lymphoepithelioma-like carcinoma of the lung

Primary lymphoepithelioma-like carcinoma (LELC) of the lung is an extremely rare disease that occurs more commonly in Asians, and is composed of undifferentiated carcinoma with prominent lymphoid stroma. LELC is reported to be closely associated with Epstein-Barr virus (EBV) infection. A case is presented here in which bronchial brushing smears in a 70-year-old man, revealed large clusters of neoplastic cells with scant cytoplasm. The nuclei were large, hyperchromatic, of irregular contour and with prominent nucleoli. Also identified were prominent intratumoral lymphoid infiltration and brisk mitotic figures. We detected EBV-coded small RNA in situ hybridization in smears. A cytologic diagnosis of a LELC was suggested. Further evaluation and immunohistochemical studies were conducted on formalin-fixed, paraffin-embedded material. Cords or nests of large neoplastic cells with enlarged nuclei and prominent nucleoli with marked lymphoid infiltration and lymphoid stroma were identified on H&E sections. Immunohistochemically, the tumor cells showed diffuse and strong membranous staining for CK(AE1/AE3), CK5/6, CK34βE12, Napsin A and Bcl-2 but were negative for CK7, CK14, CK20, EMA, TTF-1, chromogranin A, synaptophysin and CD56. The proliferative index with MIB-1 was around 60%, and the p53 positive cells around 20%. The diagnosis of primary LELC of the lung was confirmed based on cytopathologic, histopathologic, immunohistochemical and EBER results, and a detailed systemic examination to exclude possible extrapulmonary (nasopharyngeal) origin. We report the cytopathological features of LELC of the lung and demonstrate here for the first time the positivity of the EBER with RNA-ISH method in smears with emphasis on differential diagnostic considerations.     

原发肾上腺NK/T细胞淋巴瘤皮肤侵犯一例

患者女,64岁,诊断肾上腺NK/T细胞淋巴瘤1年,四肢暗红色浸润性斑块2个月。患者1年前因剧烈腹痛就诊于杭州市中医院,腹部CT发现双侧肾上腺肿块,行左肾上腺切除术,结合免疫组化、病理,诊断为非霍奇金淋巴瘤,NK/T细胞淋巴瘤。10个月前于浙江大学医学院附属邵逸夫医院就诊,腹部CT示右侧肾上腺巨大团块。血常规、凝血功能、胸部CT检查未见明显异常     

Expression of Epstein–Barr-virus-encoded small nuclear RNA in nasopharyngeal carcinomas of Aegean Turkish patients

Nasopharyngeal carcinomas (NPC) are epithelial neoplasms which show a distinct geographical distribution and have a characteristic histology. These tumors have multifactorial etiology, including virological, environmental, and genetic components. The aim of the present study is to assess the relation between Epstein–Barr-virus (EBV) and subtypes of NPC in Aegean Turkish patients. In the present study, nasopharyngeal biopsies of 84 cases diagnosed as nasopharyngeal carcinoma, between 1998 and 2004, were reevaluated. In situ hybridization with the fluorescein-conjugated EBV-encoded small nuclear RNA (EBER) oligonucleotide probe was performed on paraffin-embedded tissue sections using an automated slide stainer system. Of 84 patients, 55 were men and 29 were women with ages ranging between 7 and 77 years (median 50, mean 46.73). Seventy-three of 84 cases were EBER positive. All of 62 cases (100.0%) with undifferentiated carcinoma, 8 of 16 (50.0%) with differentiated nonkeratinizing carcinoma, and three of six (50.0%) with keratinizing squamous cell carcinoma were EBV positive. EBER positivity was statistically significantly higher in undifferentiated carcinomas, compared to the other morphological subtypes (p = 0.000). Our results showed that all morphological subtypes of NPC are highly associated with EBV latent infection in our region, and a higher prevalence was found for the undifferentiated subtype.     

Epstein–Barr virus (EBV) and Hodgkin's disease in a multi-ethnic population in Malaysia

The Epstein–Barr virus (EBV) has been implicated as a contributing factor in the development of Hodgkin's disease. Western cases of Hodgkin's disease have shown the presence of EBV in Hodgkin and Reed–Sternberg cells in approximately 50%. We studied a total of 100 consecutive cases of Hodgkin's disease from Malaysia, with the aim to elucidate its association with EBV in a multi-ethnic Asian population. Of 34 patients (34%) less than 15 years of age (childhood), 25 had classical Hodgkin's disease (eight nodular sclerosis, 16 mixed cellularity, one lymphocyte depleted) and nine had lymphocyte predominance Hodgkin's disease. Of the 66 from patients aged 15 years and above, 33 had nodular sclerosis, 24 mixed cellularity, two lymphocyte depleted, one unclassifiable and six lymphocyte predominance Hodgkin's disease. The ethnic distribution of classical Hodgkin's disease was: Malay 23, Chinese 32 and Indian 30 (Malay:Chinese:Indian = 1:1.4:1.3), and the ethnic distribution in the 15 cases of lymphocyte predominance Hodgkin's disease was: Malay four, Chinese 10 and Indian one. Taking into account the ethnic distribution of the general population and of hospital admissions, there appears to be a significant predilection of classical Hodgkin's disease cases in ethnic Indian compared to non-Indian patients (chi-squared test, 0.025 > P > 0.01). Eighty-one cases were tested for the presence of EBV by in situ hybridization for EBV encoded RNA, and 57 cases by immunostaining for EBV latent membrane protein 1. In the younger age group, all except one of the 15 cases (nine mixed cellularity, six nodular sclerosis) showed the presence of EBV (93%). In the older age group, EBV was detected (52%) in the following proportion: 6/27 nodular sclerosis, 19/22 mixed cellularity, 1/2 lymphocyte depleted, 1/1 unclassifiable. None of the 14 cases of lymphocyte predominance Hodgkin's disease showed the presence of EBV in the Hodgkin and Reed–Sternberg cells. The findings suggest a strong association of EBV with Hodgkin's disease in Malaysians (41/67, 61%), in particular childhood cases (93%). In adults, the association with EBV is significantly higher in the mixed cellularity subtype (86%) compared with the nodular sclerosis subtype (22%).     

头颈部淋巴结转移性癌中EBER1的原位杂交检测

本研究对包括鼻咽癌７例，涎腺恶性淋巴上皮病４例和口腔各部位不同分化的鳞状细胞癌５例在内的１６例肿瘤的原发灶和其淋巴结转移灶中的ＥＢＥＲ１进行了原位杂交检测，旨在揭示头颈部转移性癌中ＥＢＶ阳性表达在判断其原发肿瘤来源中的意义。结果表明：在头颈部淋巴结转移性癌中发现非角化型的癌并伴有ＥＢＥＲ１阳性者，可提示其原发灶可能来自于鼻咽癌和涎腺恶性淋巴上皮病。     

Skp2在鼻型NK/T细胞淋巴瘤中的表达及临床意义

 目的 探讨Skp2与鼻型NK/T细胞淋巴瘤的预后及其他临床特点之间的关系。方法 收集95例鼻型NK/T细胞淋巴瘤患者临床病理学资料,用免疫组化方法检测肿瘤石蜡包埋标本的EB病毒编码RNA (EBV encoded RNA,EBER)、Skp2及Ki-67的表达情况,将Skp2阳性细胞比率超过50%的病例计入高表达组,低于50%的计入低表达组,分析患者的预后、EBER表达状态与Skp2、Ki 67表达率之间的关系。结果 在Skp2高表达组,中位生存期为20.54个月(95%CI:18.49-22.59),1年总体生存率为76%;在Skp2低表达组,中位生存期为17.54个月(95%CI:20.31 22.24),1年总体生存率为71%;在Skp2阴性组,中位生存期为个21.71个月(95%CI:11.15-16.18),1年总体生存率为91%。生存分析显示,Skp2高表达组与Skp2阴性组间差异具有统计学意义(P=0.035)。结论 Skp2的高表达可能是鼻腔NK/T细胞淋巴瘤预后的不良因素。