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Journal of Virology, December 2003, p. 13267-13274, Vol. 77, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.24.13267-13274.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Lytic Viral Replication as a Contributor to the Detection of Epstein-Barr Virus in Breast Cancer

J. Huang,1 H. Chen,2 L. Hutt-Fletcher ,3,{dagger} R. F. Ambinder,2 and S. D. Hayward1,2*

Department of Pharmacology and Molecular Sciences,1 The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland,2 School of Biological Sciences, University of Missouri-Kansas City, Kansas City, Missouri3

Received 12 May 2003/ Accepted 2 September 2003

Epstein-Barr virus (EBV) has an accepted association with the epithelial malignancy nasopharyngeal carcinoma and has also been reported in other more controversial carcinoma settings. Evaluation of EBV association with epithelial carcinomas such as breast cancer would benefit from a better understanding of the outcome of EBV infection of these cells. Cell-free preparations of a green fluorescent protein-expressing virus, BX1, were used to infect breast cancer cell lines, which were then examined for EBV gene expression and viral genome copy number. Reverse transcription-PCR analyses revealed that the cells supported a mixture of latency II and lytic EBV gene expression. Lytic Zta and BMRF1 protein expression was detected by immunohistochemistry, and DNA PCR analyses estimated an EBV copy number of 300 to 600 genomes per infected cell. Evidence for lytic EBV expression was also found in breast tissue, where reverse transcription-PCR analyses detected lytic Zta transcripts in 7 of 10 breast carcinoma tissues and 4 of 10 normal tissues from the same patients. Scattered cells immunoreactive for Zta protein were also detectable in breast carcinoma. Quantitative real-time PCR analysis of EBV-positive breast carcinoma tissues suggested that less than 0.1% of the cells contained viral genomes. We suggest that sporadic lytic EBV infection may contribute to PCR-based detection of EBV in traditionally nonvirally associated epithelial malignancies.

* Corresponding author. Mailing address: The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, School of Medicine, Bunting-Blaustein Building CRB308, 1650 Orleans St., Baltimore, MD 21231. Phone: (410) 955-2548. Fax: (410) 502-6802. E-mail: dhayward{at}jhmi.edu.

{dagger} Present address: Department of Microbiology and Immunology, Louisiana State University Health Science Center, Shreveport, La

Journal of Virology, December 2003, p. 13267-13274, Vol. 77, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.24.13267-13274.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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