Detection of ebv gene expression in reed‐sternberg cells of Hodgkin's disease

Abstract

EBV DNA has been detected by Southern blot hybridization in 20-25% of Hodgkin's disease tumor specimens and localized to the Reed-Sternberg cells by in situ hybridization. In the present investigation we used a 3H-labelled EBER I anti-sense RNA for in situ hybridization of archival formalin-fixed paraffin-embedded Hodgkin's disease specimens previously shown by Southern Blot hybridization to be EBV-positive. In 6 of 8 specimens neoplastic cells showed an intense signal in virtually all of the tumor cells. The background lymphocytes, eosinophils, plasma cells and histiocytes did not demonstrate significant hybridization. In each case hybridization tended to spare the nucleoli. Hybridization was detected in specimens with histologies of mixed cellularity and nodular sclerosis. The intensity of signal relative to background is better than that in previous studies utilizing 35S-labelled large internal repeat probes for EBV in Reed-Sternberg cells. The exposure time is one week in contrast to the 4-5 weeks reported by others for detection of EBV in Hodgkin's disease. Both increased relative intensity and shorter exposure requirements may be attributed to the very high number of EBER transcripts in the target cells. The demonstration that EBER I is expressed is consistent with a role for EBV in growth regulation of Reed-Sternberg cells and suggests that the virus is not merely a silent passenger in Hodgkin's disease.

Infection of human endothelial cells with Epstein-Barr virus.

“…Reed-Sternberg and Hodgkin's cells are known to carry EBV genomes in ,'~50% of Hodgkin's lymphoma specimens (10,37). Furthermore, some follicular dendritic cells were successfully infected with EBV in vitro (38).…”

“…The procedure was performed essentially as described (10). Modification of the technique included a 10% formalin fixation to permit the use of culture chamber slides (Nunc, Inc., Naperville, IL) from which the upper structure had been removed.…”

Assessment of the prognostic impact of the Epstein–Barr virus-encoded latent membrane protein-1 expression in Hodgkin’s disease

“…Subsequently, direct evidence of EBV in tumour tissue was established by Southern blot (Weiss et al, 1987) and polymerase chain reaction (PCR) analysis (Shibata et al, 1991), although neither method was able to localize EBV to specific cell types. Later, the development of in situ hybridization for the detection of the highly abundant EBV early RNAs (EBERs) (Glickman et al, 1988;Wu et al, 1990) and immunohistochemistry for the EBVencoded latent membrane protein-1 (LMP1) (Pallesen et al, 1991;Murray et al, 1992) enabled the localization of EBV to the malignant HRS cells. Although EBV can also be detected in rare nonmalignant 'bystander' lymphocytes within HD tissues, it is the detection of EBV within HRS cells that is necessary to establish a HD case as 'EBV-associated'.…”

Further investigation of the role of HLA-DPB1 in adult Hodgkin’s disease (HD) suggests an influence on susceptibility to different HD subtypes

“…Studies suggesting that the peak of HD cases in early adulthood HD resembles that of paralytic poliomyelitis in the pre-vaccine era (Gutensohn and Cole, 1977) support an aetiological pathway in which HD arises as a rare outcome of a common infection (Gutensohn and Cole, 1981). There is now extensive evidence to suggest that the infectious agent, at least in some cases, is Epstein-Barr virus (EBV) (Weiss et al, 1989;Wu et al, 1990;Khan et al, 1992;Jarrett, 1993;Oudejans et al, 1997). However, the ubiquitous distribution of EBV in the normal population, without accompanying disease (Miller, 1990), suggests that other factors, possibly of host origin, contribute to the aetiology of HD.…”