Abstract
Epstein-Barr virus (EBV) was the first human virus associated directly with human malignancies. During EBV infection of various host cells the double-stranded linear EBV DNA carried by the virions undergoes circularization. Since there are variable numbers of terminal repetitions (TRs) at the ends of the linear EBV genome, the resulting circular episomes enclose a variable number of TRs. Thus, in cells carrying viral episomes, the sizes of the terminal restriction enzyme fragments of EBV is affected by the number of TRs (Raab-Traub and Flynn Cell 47:883-889, 1986). Southern blot analysis revealed that in monoclonal proliferations, arising from a single cell, there was only a single band representing the joined EBV termini, whereas multiple terminal restriction enzyme fragments that differ in size were characteristic for oligoclonal or polyclonal proliferations. Using suitable probes, one can distinguish the episomal form from the linear EBV genomes that are formed during lytic EBV replication or during integration into the host genome. TR analysis is a useful tool for the determination of EBV clonality in different clinical samples and in cell lines carrying EBV genomes. A single terminal restriction enzyme fragment may indicate EBV infection at an early phase of clonal cell proliferation, whereas polyclonal EBV genomes may derive from multiple infections of proliferating cells.
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References
Raab-Traub N, Flynn K (1986) The structure of the termini of the Epstein-Barr virus as a marker of clonal cellular proliferation. Cell 47(6):883–889
Zimmermann J, Hammerschmidt W (1995) Structure and role of the terminal repeats of Epstein-Barr virus in processing and packaging of virion DNA. J Virol 69(5):3147–3155
Moody CA, Scott RS, Su T, Sixbey JW (2003) Length of Epstein-Barr virus termini as a determinant of epithelial cell clonal emergence. J Virol 77(15):8555–8561
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98(3):503–517
Feederle R, Shannon-Lowe C, Baldwin G, Delecluse HJ (2005) Defective infectious particles and rare packaged genomes produced by cells carrying terminal-repeat-negative Epstein-Barr virus. J Virol 79(12):7641–7647. doi:10.1128/JVI.79.12.7641-7647.2005
Chiu SH, Wu MC, Wu CC, Chen YC, Lin SF, Hsu JT et al (2014) Epstein-Barr virus BALF3 has nuclease activity and mediates mature virion production during the lytic cycle. J Virol 88(9):4962–4975. doi:10.1128/JVI.00063-14
Gulley ML, Raphael M, Lutz CT, Ross DW, Raab-Traub N (1992) Epstein-Barr virus integration in human lymphomas and lymphoid cell lines. Cancer 70(1):185–191
Fan H, Gulley ML (2001) Molecular methods for detecting Epstein-Barr virus (part ii): structural analysis of Epstein-Barr virus DNA as a marker of clonality. Methods Mol Med 49:313–319. doi:10.1385/1-59259-081-0:313
Lewin N, Minarovits J, Weber G, Ehlin-Henriksson B, Wen T, Mellstedt H et al (1991) Clonality and methylation status of the Epstein-Barr virus (EBV) genomes in in vivo-infected EBV-carrying chronic lymphocytic leukemia (CLL) cell lines. Int J Cancer 48(1):62–66
Gulley ML, Eagan PA, Quintanilla-Martinez L, Picado AL, Smir BN, Childs C et al (1994) 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 83(6):1595–1602
Gulley ML, Pulitzer DR, Eagan PA, Schneider BG (1996) Epstein-Barr virus infection is an early event in gastric carcinogenesis and is independent of bcl-2 expression and p53 accumulation. Hum Pathol 27(1):20–27
van de Rijn M, Cleary ML, Variakojis D, Warnke RA, Chang PP, Kamel OW (1996) Epstein-Barr virus clonality in lymphomas occurring in patients with rheumatoid arthritis. Arthritis Rheum 39(4):638–642
Lin CT, Chen W, Hsu MM, Dee AN. Clonal versus polyclonal Epstein-Barr virus infection in nasopharyngeal carcinoma cell lines. Laboratory investigation; a journal of technical methods and pathology. 1997;76(6):793-8.
Ryan JL, Kaufmann WK, Raabtraub N, Oglesbee SE, Carey LA, Gulley ML (2006) Clonal evolution of lymphoblastoid cell lines. Lab Invest 86(11):1193–200. doi:10.1038/labinvest.3700472
Arai A, Yamaguchi T, Komatsu H, Imadome K, Kurata M, Nagata K et al (2014) Infectious mononucleosis accompanied by clonal proliferation of EBV-infected cells and infection of CD8-positive cells. Int J Hematol 99(5):671–675. doi:10.1007/s12185-014-1548-4
Brown NA, Liu CR, Wang YF, Garcia CR (1988) B-cell lymphoproliferation and lymphomagenesis are associated with clonotypic intracellular terminal regions of the Epstein-Barr virus. J Virol 62(3):962–969
Raab-Traub N, Rajadurai P, Flynn K, Lanier AP (1991) Epstein-Barr virus infection in carcinoma of the salivary gland. J Virol 65(12):7032–7036
Pathmanathan R, Prasad U, Chandrika G, Sadler R, Flynn K, Raab-Traub N (1995) Undifferentiated, nonkeratinizing, and squamous cell carcinoma of the nasopharynx. Variants of Epstein-Barr virus-infected neoplasia. Am J Pathol 146(6):1355–1367
Pathmanathan R, Prasad U, Sadler R, Flynn K, Raab-Traub N (1995) Clonal proliferations of cells infected with Epstein-Barr virus in preinvasive lesions related to nasopharyngeal carcinoma. N Engl J Med 333(11):693–698. doi:10.1056/NEJM199509143331103
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Bánáti, F., Koroknai, A., Szenthe, K. (2017). Terminal Repeat Analysis of EBV Genomes. In: Minarovits, J., Niller, H. (eds) Epstein Barr Virus. Methods in Molecular Biology, vol 1532. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6655-4_12
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DOI: https://doi.org/10.1007/978-1-4939-6655-4_12
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