Analysis of the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) gene and promoter in Hodgkin's disease isolates: selection against EBV variants with mutations in the LMP-1 promoter ATF-1/CREB-1 binding site.
OBJECTIVE
To study the distribution of Epstein-Barr virus (EBV) variants containing mutations in the latent membrane protein 1 (LMP-1) oncogene and promoter in EBV associated Hodgkin's disease and infectious mononucleosis compared with previous findings in asymptomatic EBV carriers.


METHODS
Sequence analysis of the EBV LMP-1 promoter and gene in isolates from Danish patients with Hodgkin's disease (n = 61) and infectious mononucleosis (n = 10).


RESULTS
Viruses (previously designated group D) that contain two mutations in the activating transcription factor/cAMP response element (ATF/CRE) in the LMP-1 promoter, which are known to decrease promoter activity greatly, were significantly less frequent in Hodgkin's disease than in both infectious mononucleosis (p = 0.0081) and asymptomatic EBV carriers (p = 0.0084). In some cases, the LMP-1 gene contained mutations in a recently identified cytotoxic T cell (CTL) epitope. Most viral isolates contained mutations shown to increase nuclear factor kappa B (NF-kappa B) activation and had one of two newly identified C-terminal activation regions 3 (CTAR-3) deleted. The exon 1 Xho-I restriction site in the LMP-1 gene could be lost through a range of different mutations.


CONCLUSIONS
These findings indicate selection pressure against EBV strains with weak LMP-1 promoter activity in Hodgkin's disease and thus provide further strong circumstantial evidence for the pathogenic role of EBV (and LMP-1) in this disease. Mutation of the CTL epitope suggests immune selection of EBV strains. Many EBV isolates contain functionally important mutations in the LMP-1 gene. Loss of the Xho-I restriction site should not be used as a marker of specific LMP-1 variants.
Sandvej
K
K
Laboratory of Immunopathology, Kommunehospitalet, Aarhus, Denmark. Kristian.Sandvej@aas.auh.dk
Andresen
B S
BS
Zhou
X G
XG
Gregersen
N
N
Hamilton-Dutoit
S
S
D051697:Activating Transcription Factor 1; D001483:Base Sequence; D050882:CREB-Binding Protein; D004268:DNA-Binding Proteins; D018984:Epitopes, T-Lymphocyte / Q000235:genetics; D005819:Genetic Markers; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006689:Hodgkin Disease / Q000276:immunology / Q000821:virology*; D006801:Humans; D007244:Infectious Mononucleosis / Q000276:immunology / Q000821:virology*; D008969:Molecular Sequence Data; D009154:Mutation; D016328:NF-kappa B / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013602:T-Lymphocytes, Cytotoxic / Q000648:ultrastructure; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014763:Viral Matrix Proteins / Q000235:genetics*
D051697:Activating Transcription Factor 1; D004268:DNA-Binding Proteins; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D018984:Epitopes, T-Lymphocyte; D005819:Genetic Markers; D016328:NF-kappa B; D009687:Nuclear Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014763:Viral Matrix Proteins; D050882:CREB-Binding Protein; C494363:CREBBP protein, human