Epstein–Barr virus in Reed–Sternberg G-like cells in non Hodgkin's lymphomas

In the course of our study on Hodgkin's disease (HD), ten cases of non-Hodgkin's lymphomas (NHL) containing Hodgkin and Reed-Sternberg-like (MRS) cells were encountered. Many of these cases had initially been diagnosed as HD, but on careful review of the histology, with the aid of immunophenotyping studies, they were reclassified as NHL. The presence of Epstein–Barr virus (EBV) in these HRS-like cells was investigated using a combination of EBER in situ hybridization (ISH) and immunostaining for the detection of EBV-encoded latent membrane protein (LMP). HRS-like cells in four cases (two lymphoplasmacytoid lymphomas, one Richter's transformation of lymphoplasmacytoid lymphoma, and one immunoblastic lymphoma of T-cell type) were found to be EBV-positive. In two of these cases, a second biopsy taken up to 10 years later also contained EBV in the HRS-like cells. In three of the four cases, HRS-like cells expressed the activation antigen CD30, but the expression of B- or T-cell antigens was variable. All cases of T-cell-rich B-cell lymphomas were negative for EBV. In conclusion, EBV may play a role in the development of HRS-like cells i some cases of NHL. The relationship of HRS-like cells to HRS cells of HD is discussed.     

Incidence of latent infection of Epstein–Barr virus in lung cancers—an analysis of EBER1 expression in lung cancers by in situ hybridization

To evaluate the presence of Epstein–Barr virus (EBV) in lung cancers of Japanese patients, 81 lung cancers were examined using a highly sensitive in situ hybridization (ISH) method, employing an antisense oligonucleotide probe for EBV-encoded small nuclear RNA-1 (EBER). EBER1 expression was demonstrated in one poorly differentiated squamous cell carcinoma associated with marked lymphoid stroma (PDSCC-LS), two well differentiated adenocarcinomas, and two moderately differentiated squamous cell carcinomas, but was not detectable in other lung cancers, including small cell carcinomas. Unlike lymphoepithelioma-like undifferentiated carcinoma (LELC) of the lung, the PDSCC-LS consisted of poorly differentiated cells with distinct cell borders and nuclei with a coarse chromatin pattern and some prominent nucleoli. Most of the cancer cells expressed intense EBER1 signals. Although small to moderate numbers of cells positive for EBER1 were present in two adenocarcinomas and two squamous cell carcinomas, EBER1 signals varied in intensity and number in these four cases. Although polymerase chain reaction (PCR) and Southern blot hybridization with a ³²P-labelled probe internal to the primers were conducted to detect the EBV genome in 24 lung cancers, including five EBER1-positive cases, the genome was found to be positive in the five cases with EBER1-positive staining, including the PDSCC-LS, two adenocarcinomas and two squamous cell carcinomas, but not in the other cases. This study indicates that the morphological features of EBV-associated lung cancers are not restricted to the typical LELC type.     

非特指性外周T细胞淋巴瘤中EBER、 LMP1的表达及与预后的关系

目的 探讨非特指性外周T细胞淋巴瘤(peripheral T-cell lymphoma,not otherwise specified,PTCL-NOS)中EBER、LMP1的表达及与患者预后的关系.方法 采用原位分子杂交(in situ hybridization,ISH)技术和免疫组化法分别检测81例PTCL-NOS及59例对照组[48例血管免疫母细胞性T细胞淋巴瘤(angioimmunoblastic T-cell lymphoma,AITL)和11例结外NK/T细胞淋巴瘤(extranodal NK/T cell lymphoma,ENK/TCL)]中EBER和LMP1的表达,并分析EBER表达与PTCL-NOS患者临床病理特征及预后的关系.结果 (1)81例PTCL-NOS中,EBER阳性率为43.2％ (35/81);35例EBER阳性的PTCL-NOS病例中免疫组化得分1分+2分者共29例,占EBER阳性病例的82.9％(29/35),3分+4分者共6例,占17.1％ (6/35).EBER表达与PTCL-NOS患者年龄、性别、乳酸脱氢酶(lactate dehydrogenase,LDH)水平及临床分期均无明显相关性(P＞0.05).(2)81例PTCL-NOS组织中,LMP1蛋白阳性率为22.2％(18/81).LMP1蛋白表达与EBER表达具有一致性,但EBER阳性率明显高于LMP1 (P ＜0.05).(3)33例PTCL-NOS获得临床随访资料,随访时间1～63个月,中位生存期为23个月,总生存率为33.3％(11/33).Kaplan-Meier生存曲线分析显示,EBER阳性组的生存率明显低于EBER阴性组(P＜0.05).结论 EB病毒(EBV)感染可能是PTCL-NOS发生、发展中重要但非根本性的因素.EBER-ISH检测EBV感染具有较高的敏感性和特异性.EBV感染对PTCL-NOS患者的预后判断具有重要意义.     

Vascular lesion in a patient of chronic active Epstein-Barr virus infection with hypersensitivity to mosquito bites: vasculitis induced by mosquito bite with the infiltration of nonneoplastic Epstein-Barr virus-positive cells and subsequent development of natural killer/T-cell lymphoma with angiodestruction

This report describes a vasculitis and subsequently developing angiodestructive lymphoma in an 11-year-old Japanese-Filipino girl exhibiting mosquito allergy with the background of chronic active Epstein-Barr virus (EBV) infection. She developed necrotic skin ulcer at the site of mosquito bite, and histopathological examination revealed EBV-positive mononuclear cell infiltration throughout the wall of small-sized muscular artery. These EBV-positive lymphoid cells were oligoclonal in Southern blot analysis for EBV terminal repeats. Effectiveness of steroid therapy also supports the nonneoplastic nature. Approximately 1 year later, she developed progressive large skin ulcer without mosquito bites. Microscopically, the angiocentric or angiodestructive pattern of EBV-positive atypical cells supported the diagnosis of extranodal natural killer/T-cell lymphoma. Southern blot analysis revealed the monoclonal neoplastic nature of EBV-positive cells. In contrast to the primary mosquito bite lesion, natural killer/T-cell lymphoma cells exhibited the higher expression of EBV latent membrane protein 1 mRNA and the apparent protein expression detected by immunohistochemistry.     

Evidence of Epstein-Barr virus infection in ulcerative colitis

BACKGROUND AND AIM: The aetiology of ulcerative colitis is still controversial, however, recent studies have emphasised the possible role of infectious agents or ingested substances and their breakdown products, which might activate immune-mediated mechanisms eventually leading to tissue damage. Aim of this investigation was to ascertain the occurrence and the potential role of Epstein-Barr virus infection in large bowel mucosa of ulcerative colitis patients. PATIENTS AND METHODS: Twenty-three biopsies and six total colectomies from 17 patients were analysed for the expression of Epstein-Barr virus proteins and RNAs. Polymerase chain reaction experiments were also carried out to detect Epstein-Barr virus DNA. For comparison, ten biopsies from patients with Crohn's disease, ten biopsies from patients with different types of colitis, seven biopsies and five surgical margins of normal colonic mucosa from the small and large bowels were studied (controls). RESULTS: Six biopsies and four colectomies from seven ulcerative colitis patients showed scattered lymphocytes expressing nuclear EBER 1-2 and harbouring polymerase chain reaction-amplifiable Epstein-Barr virus-DNA. In some cases, linear viral DNA (typical of lytic Epstein-Barr virus infection) was also found. Epithelial cells were invariably negative in all cases. All control tissues from non-ulcerative colitis patients were also invariably non-reactive. CONCLUSION: Evidence of Epstein-Barr virus infection in the mucosal inflammatory cells of ulcerative colitis patients suggests a possible role of this virus in the chronicity of ulcerative colitis.     

Survivin在中线T细胞淋巴瘤中的表达及其与EB病毒感染的关系

为了探讨凋亡抑制基因存活蛋白（survivin）在中线T细胞淋巴瘤（midline T-cell lymphoma,MTL）中的表达及其与EB病毒（Epstin-Barr virus,EBV）感染的关系,应用免疫组织化学方法检测石蜡切片中存活蛋白和EBV潜伏膜蛋白（latent membrane protein,LMP-1）的表达,采用原位分子杂交技术检测EBV编码的RNA（EBV-encoded RNA,EBER1/2）.结果表明：41例MTL中26例存活蛋白阳性表达（63.42%）,而在10例反应性增生淋巴组织中均未检测出存活蛋白表达;存活蛋白在低度恶性MTL中的阳性表达率为12.50%,在中-高度恶性MTL中的阳性表达率为75.76%,两组之间的差异具有显著性的意义（x^2=8.55,P＜0.01）;EBER 1/2阳性率为70.73%,LMP-1阳性率为41.46%,EBV阳性组与EBV阴性组之间存活蛋白的表达差异无显著性意义（P＞0.05）.结论：MTL组织中存活蛋白表达上调,存活蛋白表达阳性率与MTL的恶性程度有关,存活蛋白基因可能通过肿瘤细胞凋亡的抑制参与MTL的发生发展,但未发现存活蛋白表达与EBV感染之间有明显相关性.     

Analysis of Epstein–Barr virus (EBV) type and variant in spontaneous lymphoblastoid cells and Hu-SCID mouse tumours

Epstein–Barr virus (EBV) type and strain variations were examined using both lymphoblastoid cell lines (LCLs), spontaneously derivedin vitrofrom peripheral blood mononuclear cells (PBMC) of 15 HIV-1-seropositive individuals, and SCID mouse tumours induced by inoculation of PBMC from 11 healthy human donors (Hu-SCID tumours). Polymerase chain reaction (PCR) analysis disclosed that all but one of the 26 EBV+ samples harboured EBV nuclear antigen (EBNA) 2 and 3C type A virus. On the other hand, single strand conformation polymorphism (SSCP) analysis using Epstein–Barr encoded RNA (EBER) specific primers detected an AG876-like (type B) band pattern in 21 of the 26 EBV+ samples. Three Hu-SCID tumours scored as B95.8-like (type A), and two showed neither a type A nor a type B SSCP migration pattern. Sequence analysis of the amplified EBER fragments confirmed the PCR-SSCP findings; moreover, additional mutations were present not only in the two EBV+ samples with anomalous SSCP pattern, but also in two other samples with a standard SSCP profile. Thus, EBER analysis did not correlate with EBNA typing, and appeared to be unsuitable for EBV type assessment. Latent membrane protein (LMP) analysis disclosed, on the whole, seven size variants: as expected, the differences were due to the variable numbers of a 33-bp repeat in the amplified fragment, as assessed by direct sequencing. The broader variability detected by LMP analysis should prove more useful than typing for assessing the presence of single and/or mixed variants resulting from EBV reactivation and/or reinfection.     

炎性假瘤样滤泡树突细胞肉瘤的临床病理学特征

目的探讨炎性假瘤样滤泡树突细胞肉瘤(IPT-like FDCS)的临床病理学特征及其鉴别诊断。方法运用组织病理学、免疫组织化学染色,回顾性分析3例IPT-like FDCS的特点,并结合相关文献进行讨论。结果肿瘤由圆形、卵圆形及梭形细胞组成,呈编织状、漩涡状或席纹状排列,与周围组织分界清。肿瘤细胞呈合体状,染色质细颗粒状,可见单个小核仁,细胞轻度异型,其间见多量淋巴细胞、浆细胞混杂。免疫组化结果:肿瘤细胞不同程度地表达CD21、CD35、CD23滤泡树突细胞标记物,伴随IgG4浆细胞局灶性高表达。EBER原位杂交显示瘤细胞散在阳性。结论IPT-like FDCS是一种好发于女性的少见低度恶性肿瘤,与EB病毒感染有关,最常见的发病部位是脾脏和肝脏,对其认识不足时很容易误诊或漏诊。     

Analysis of Epstein-Barr virus in hodgkin's disease: Experience of a single university hospital in Korea

Hodgkin's disease is known to be associated with Epstein-Barr virus (EBV) infection in Western countries, and viral nucleic acids and proteins have been identified within Reed-Sternberg (RS) cells, which are the histopathologic hallmark of the disease process. Twenty-five cases of Hodgkin's disease from a single university hospital in Korea were studied for evidence of EBV by in situ hybridization for EBV DNA and RNA and immunohistochemistry for an EBV latent protein. EBV nucleic acids were studied by a rapid (60 minutes) in situ hybridization procedure, which utilized biotinylated DNA probes specific for the following nucleic acid sequences: (1) EBV EBER1 RNA (an abundant RNA sequence expressed during latent EBV infection), (2) EBV Notl repeats (a tandemly repeated DNA sequence, which has been established to identify amplified EBV genome in lytic EBV infection), and (3) BAM HI W (a DNA sequence reiterated 11 times within the viral genome). In addition, immunohistochemistry for EBV latent membrane protein, a protein that is capable of inducing cellular transformation in cell culture, was also performed. EBV was identified within the neoplastic RS cells by at least one method in 19/25 cases (76%). The mixed cellularity subtype was the most common subtype associated with EBV infection (11/13–85%). In situ hybridization for EBV EBER1 RNA was the most sensitive method for EBV detection and was present in 17/25 cases. A significant proportion of Korean Hodgkin's disease cases is associated with EBV infection. © 1994 Wiley-Liss, Inc.