Association of Epstein-Barr virus genome with mixed cellularity and cellular phase nodular sclerosis Hodgkin's disease subtypes

Recent studies have suggested that Epstein-Barr virus (EBV) may play a role in the etiology of Hodgkin's disease (HD). In a previous study, we used a latent membrane protein 1 (LMP1)-specific antibodies to examine archival material from 74 British children with HD and found 50% of cases to be positive. It is known that there are geographic and ethnic variations in the incidence of HD. We have investigated LMP1 status in formalin-fixed, paraffin wax-embedded lymph nodes with HD involvement from 53 children and 48 adults from Kenya using immunohistochemical staining. We also developed sensitive and specific in vitro gene amplification protocols for examining the EBV strain type in such material using several combinations of primers derived from the EBNA 2 and EBNA 3 coding regions. LMP1 positivity was present in 100% of the pediatric cases (two lymphocyte-predominant, 25 nodular sclerosis, 16 mixed cellularity, 5 lymphocyte depletion, and 5 unclassified) and in 66% of the adult cases (two of three lymphocyte-predominant, 26 of 39 nodular, sclerosis, two of two mixed cellularity, and two of four lymphocyte depletion). Tests to type the EBV strain were undertaken in 25 EBV-positive pediatric cases. A combination of type-specific polymerase chain reactions for EBNA 2 and EBNA 3C genes indicated that seven patients had type 1, eight had type 2, and 10 had dual infections with both types. Five cases with dual infections were further investigated using a sensitive in situ hybridization for the EBV- encoded, small nuclear nonpolyadenylated RNAs (EBERs). EBER transcripts were detected in Reed-Sternberg and Hodgkin cells and in occasional infiltrating lymphocytes. These observations indicate that in Kenya EBV is consistently associated with pediatric cases of HD, and that biopsies from a number of such cases appear to carry both type 1 and type 2 viral sequences.

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Epstein-Barr virus DNA is abundant and monoclonal in the Reed-Sternberg cells of Hodgkin's disease: association with mixed cellularity subtype and Hispanic American ethnicity

Blood ◽

10.1182/blood.v83.6.1595.bloodjournal8361595 ◽

1994 ◽

Vol 83 (6) ◽

pp. 1595-1602 ◽

Cited By ~ 1

Author(s):

ML Gulley ◽

PA Eagan ◽

L Quintanilla-Martinez ◽

AL Picado ◽

BN Smir ◽

...

Keyword(s):

Hodgkin's Disease ◽

Odds Ratio ◽

Epstein Barr Virus ◽

Hodgkin’S Disease ◽

Viral Dna ◽

Mixed Cellularity ◽

Hispanic Ethnicity ◽

Barr Virus ◽

Ebv Dna ◽

Epstein Barr

One hundred twenty-five cases of Hodgkin's disease from the United States (79), Mexico City (31), and Costa Rica (15) were analyzed for the presence of Epstein-Barr virus (EBV) by in situ hybridization to EBER1 transcripts. EBV was more frequently detected in the Reed- Sternberg (RS) cells of mixed cellularity Hodgkin's disease (37 of 48 [77%]) compared with the nodular sclerosis subtype (19 of 71 [27%], P < .001). The presence of EBV was also associated with Hispanic ethnicity (P < .001). In a multivariate analysis, patient age, gender, and geographic location were less predictive of EBV positivity than were mixed cellularity histology (odds ratio = 8.3) and Hispanic ethnicity (odds ratio = 4.3). Southern blot analysis of EBV terminal repeat fragments using the Xho1a probe showed that the viral DNA was monoclonal in 17 of 17 cases having EBER1-positive RS cells. By comparison, EBV DNA was not detected by Southern analysis in 20 cases lacking EBER1 in RS cells, even when occasional background lymphocytes expressed EBER1. Because clonal viral DNA was so readily detected in EBER1-positive cases, the EBV genome is probably amplified at least 50- fold in the infected RS cells. Monoclonality of EBV DNA implies that the RS cells were infected before malignant transformation.

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Presence of Epstein-Barr virus and strain type assignment in Argentine childhood Hodgkin's disease

Blood ◽

10.1182/blood.v86.10.3922.bloodjournal86103922 ◽

1995 ◽

Vol 86 (10) ◽

pp. 3922-3929 ◽

Cited By ~ 29

Author(s):

MV Preciado ◽

E De Matteo ◽

B Diez ◽

J Menarguez ◽

S Grinstein

Keyword(s):

Hodgkin's Disease ◽

Epstein Barr Virus ◽

Polymerase Chain Reaction Amplification ◽

Hodgkin’S Disease ◽

Nuclear Antigen ◽

Mixed Cellularity ◽

Barr Virus ◽

Type Assignment ◽

Epstein Barr ◽

Age Range

Epstein-Barr virus (EBV) has been implicated in the etiology of a large number of malignancies. Most recently several studies have linked EBV to Hodgkin's disease. In this report, formalin-fixed, paraffin-embedded tissues were collected retrospectively from 41 children with Hodgkin's disease treated at our hospital. Lymph node biopsies were examined for the presence of two virus-encoded latent proteins: latent membrane protein (LMP) and Epstein-Barr nuclear antigen-2 (EBNA-2), in Reed- Sternberg (RS) and Hodgkin (H) cells, by peroxidase immunolabeling. Nonisotopic Epstein-Barr encoded RNAs (EBERs) in situ hybridization was also performed and positive labeling in malignant cells was detected. Twenty specimens were EBER+/LMP+, 2 were EBER+/LMP-, and 19 were EBER- /LMP-. However, none of the 41 cases expressed EBNA-2. Twenty-two of 41 (54%) cases were EBV positive including 2 of 6 with lymphocyte predominance, 19 of 25 with mixed cellularity, 0 of 9 with nodular sclerosis, and 1 of 1 with lymphocyte depletion. In the age range of 2 to 6 years, 14 of 17 (82%) samples were EBV-positive, whereas only 8 of 24 (33%) samples from the age range of 7 to 15 years contained EBV. (P = .004), a two-tailed Fisher's test). In 17 samples, polymerase chain reaction amplification was performed using strain specific primers for exon sequences of the EBNA-3C gene of EBV. From 12 positive samples, 8 contained EBV-A and 4 EBV-B. These results support the hypothesis that EBV contributes to the pathogenesis of pediatric Hodgkin's disease, particularly in mixed cellularity Hodgkin's disease and in the younger group.

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Rearranged Epstein-Barr Virus Genome in Hodgkin's Disease and Angioimmunoblastic Lymphadenopathy: Swiss Results

American Journal Of Pathology ◽

10.1016/s0002-9440(10)63662-0 ◽

2003 ◽

Vol 163 (1) ◽

pp. 369-370 ◽

Cited By ~ 2

Author(s):

Hans Knecht ◽

Bernhard F. Odermatt

Keyword(s):

Hodgkin's Disease ◽

Epstein Barr Virus ◽

Hodgkin’S Disease ◽

Virus Genome ◽

Angioimmunoblastic Lymphadenopathy ◽

Barr Virus ◽

Epstein Barr

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Epstein-Barr virus DNA is abundant and monoclonal in the Reed-Sternberg cells of Hodgkin's disease: association with mixed cellularity subtype and Hispanic American ethnicity

Blood ◽

10.1182/blood.v83.6.1595.1595 ◽

1994 ◽

Vol 83 (6) ◽

pp. 1595-1602 ◽

Cited By ~ 103

Author(s):

ML Gulley ◽

PA Eagan ◽

L Quintanilla-Martinez ◽

AL Picado ◽

BN Smir ◽

...

Keyword(s):

Hodgkin's Disease ◽

Odds Ratio ◽

Epstein Barr Virus ◽

Hodgkin’S Disease ◽

Viral Dna ◽

Mixed Cellularity ◽

Hispanic Ethnicity ◽

Barr Virus ◽

Ebv Dna ◽

Epstein Barr

Abstract One hundred twenty-five cases of Hodgkin's disease from the United States (79), Mexico City (31), and Costa Rica (15) were analyzed for the presence of Epstein-Barr virus (EBV) by in situ hybridization to EBER1 transcripts. EBV was more frequently detected in the Reed- Sternberg (RS) cells of mixed cellularity Hodgkin's disease (37 of 48 [77%]) compared with the nodular sclerosis subtype (19 of 71 [27%], P < .001). The presence of EBV was also associated with Hispanic ethnicity (P < .001). In a multivariate analysis, patient age, gender, and geographic location were less predictive of EBV positivity than were mixed cellularity histology (odds ratio = 8.3) and Hispanic ethnicity (odds ratio = 4.3). Southern blot analysis of EBV terminal repeat fragments using the Xho1a probe showed that the viral DNA was monoclonal in 17 of 17 cases having EBER1-positive RS cells. By comparison, EBV DNA was not detected by Southern analysis in 20 cases lacking EBER1 in RS cells, even when occasional background lymphocytes expressed EBER1. Because clonal viral DNA was so readily detected in EBER1-positive cases, the EBV genome is probably amplified at least 50- fold in the infected RS cells. Monoclonality of EBV DNA implies that the RS cells were infected before malignant transformation.

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Epstein-Barr virus and childhood Hodgkin's disease in Honduras and the United States

Blood ◽

10.1182/blood.v81.2.462.462 ◽

1993 ◽

Vol 81 (2) ◽

pp. 462-467 ◽

Cited By ~ 144

Author(s):

RF Ambinder ◽

PJ Browning ◽

I Lorenzana ◽

BG Leventhal ◽

H Cosenza ◽

...

Keyword(s):

United States ◽

Hodgkin's Disease ◽

Epstein Barr Virus ◽

Hodgkin’S Disease ◽

The United States ◽

Mixed Cellularity ◽

Detection Techniques ◽

Barr Virus ◽

Epstein Barr ◽

Initial Peak

Abstract In industrialized populations, Hodgkin's disease (HD) has an initial peak in young adulthood, whereas in economically developing populations the initial peak occurs in childhood. This pattern resembles that of infection with poliovirus and suggests an infectious cofactor in the etiology. Serologic studies have linked Epstein-Barr virus (EBV) to young adult and adult HD, and viral nucleic acids and antigens have been detected in a subset of Hodgkin's tumor specimens. To investigate the association of childhood HD with EBV we studied tumor specimens from 11 children treated in Honduras and 25 children treated in the United States using in situ hybridization and antigen detection techniques. Among the patients from Honduras, tumor specimens from all cases were EBV positive. Among the patients from the United States, tumor specimens from six of seven patients with mixed cellularity histology, 2 of 15 with nodular sclerosis histology, and neither of two patients with lymphocyte-predominant histologies were EBV positive. These findings support the hypothesis that EBV contributes to the pathogenesis of HD in children, particularly in mixed cellularity HD, and raises the possibility that there are important geographic, racial, or ethnic factors in the EBV association with HD.

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