This Article Order Full text via Infotrieve Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Oudejans, J. J. Articles by Meijer, C. J. Search for Related Content PubMed PubMed Citation Articles by Oudejans, J. J. Articles by Meijer, C. J.

American Journal of Pathology, Vol 147, 923-933, Copyright © 1995 by American Society for Investigative Pathology

REGULAR ARTICLES

Detection of heterogeneous Epstein-Barr virus gene expression patterns within individual post-transplantation lymphoproliferative disorders

JJ Oudejans, M Jiwa, AJ van den Brule, FA Grasser, A Horstman, W Vos, PM Kluin, P van der Valk, JM Walboomers and CJ Meijer
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.

Using RT-PCR analysis of Epstein-Barr virus (EBV) latent gene transcription in EBV-harboring cell lines (JY and RAJI) and in post- transplantation lymphoproliferative disorders (PT-LPDs), we detected transcription of all tested latent genes (EBNA1, EBNA2, LMP1, LMP2A, and BARF0) in all cases, suggesting the presence of similar EBV expression patterns in both PT-LPDs and cell lines. In addition, the detection of immediate early (ZEBRA) and early gene (BHRF1) transcripts in cell lines and PT-LPDs indicates that activation of the virus lytic cycle occurs. To investigate EBV expression patterns at the single-cell level, a combination of immunohistochemistry and RNA in situ hybridization (including double-staining procedures) was used. In the JY and RAJI cell lines, the latency type 3 expression pattern was detected in 80 to 90% of the cells as shown by the co-expression of EBNA2 and LMP1. In contrast, in the three PT-LPDs that could be analyzed by double staining, cells expressing both EBNA2 and LMP1 were rarely detected. A mixture of at least three different cell populations were identified: (1) cells exclusively expressing EBER1/2 and EBNA1 (latency type 1); (2) cells expressing EBER1/2, EBNA1, and LMP1 (latency type 2); and (3) cells expressing EBER1/2, EBNA1, and EBNA2 in the absence of LMP1. Activation of the lytic cycle was observed in a small minority of cells, as demonstrated by detection of ZEBRA and EA-D in all cases and GP350/220 in two cases. Thus, in contrast to EBV- transformed cell lines, the observed EBV gene expression pattern in PT- LPDs reflects a mixture of multiple EBV-harboring subpopulations expressing different subsets of EBV-encoded proteins. These data indicate that the operational definitions of EBV latencies in vitro cannot easily be applied to PT-LPDs but that a continuum of different latency expression patterns can be detected at the single cell level in these lymphomas with, in a small minority of cells, progression to the virus lytic cycle.

This article has been cited by other articles:

				L. L. Kis, M. Takahara, N. Nagy, G. Klein, and E. Klein IL-10 can induce the expression of EBV-encoded latent membrane protein-1 (LMP-1) in the absence of EBNA-2 in B lymphocytes and in Burkitt lymphoma- and NK lymphoma-derived cell lines Blood, April 1, 2006; 107(7): 2928 - 2935. [Abstract] [Full Text] [PDF]

				P. S. Beisser, D. Verzijl, Y. K. Gruijthuijsen, E. Beuken, M. J. Smit, R. Leurs, C. A. Bruggeman, and C. Vink The Epstein-Barr Virus BILF1 Gene Encodes a G Protein-Coupled Receptor That Inhibits Phosphorylation of RNA-Dependent Protein Kinase J. Virol., January 1, 2005; 79(1): 441 - 449. [Abstract] [Full Text] [PDF]

				A. zur Hausen, J. van Beek, E. Bloemena, F. J. ten Kate, C. J. L. M. Meijer, and A. J. C. van den Brule No role for Epstein-Barr virus in Dutch hepatocellular carcinoma: a study at the DNA, RNA and protein levels J. Gen. Virol., July 1, 2003; 84(7): 1863 - 1869. [Abstract] [Full Text] [PDF]

				R. Loomis, R. Carbone, M. Reiss, and J. Lacy Bcl-2 Antisense (G3139, Genasense) Enhances the in Vitro and in Vivo Response of Epstein-Barr Virus-associated Lymphoproliferative Disease to Rituximab Clin. Cancer Res., May 1, 2003; 9(5): 1931 - 1939. [Abstract] [Full Text] [PDF]

				J. Kurth, M.-L. Hansmann, K. Rajewsky, and R. Kuppers Epstein-Barr virus-infected B cells expanding in germinal centers of infectious mononucleosis patients do not participate in the germinal center reaction PNAS, April 15, 2003; 100(8): 4730 - 4735. [Abstract] [Full Text] [PDF]

				P. Porcu, C. F. Eisenbeis, R. P. Pelletier, E. A. Davies, R. A. Baiocchi, S. Roychowdhury, S. Vourganti, G. J. Nuovo, W. L. Marsh, A. K. Ferketich, et al. Successful treatment of posttransplantation lymphoproliferative disorder (PTLD) following renal allografting is associated with sustained CD8+ T-cell restoration Blood, September 18, 2002; 100(7): 2341 - 2348. [Abstract] [Full Text] [PDF]

				I. Johannessen, S. M. Perera, A. Gallagher, P. A. Hopwood, J. A. Thomas, and D. H. Crawford Expansion in scid mice of Epstein-Barr virus-associated post-transplantation lymphoproliferative disease biopsy material J. Gen. Virol., January 1, 2002; 83(1): 173 - 178. [Abstract] [Full Text] [PDF]

				A A T P Brink, C J L M Meijer, J M Nicholls, J M Middeldorp, and A J C van den Brule Activity of the EBNA1 promoter associated with lytic replication (Fp) in Epstein-Barr virus associated disorders Mol. Pathol., April 1, 2001; 54(2): 98 - 102. [Abstract] [Full Text]

				S. J. C. Stevens, E. A. M. Verschuuren, I. Pronk, W. van der Bij, M. C. Harmsen, T. H. The, C. J. L. M. Meijer, A. J. C. van den Brule, and J. M. Middeldorp Frequent monitoring of Epstein-Barr virus DNA load in unfractionated whole blood is essential for early detection of posttransplant lymphoproliferative disease in high-risk patients Blood, March 1, 2001; 97(5): 1165 - 1171. [Abstract] [Full Text] [PDF]

				A. z. Hausen, A. A. T. P. Brink, M. E. Craanen, J. M. Middeldorp, C. J. L. M. Meijer, and A. J. C. van den Brule Unique Transcription Pattern of Epstein-Barr Virus (EBV) in EBV-carrying Gastric Adenocarcinomas: Expression of the Transforming BARF1 Gene Cancer Res., May 1, 2000; 60(10): 2745 - 2748. [Abstract] [Full Text]

				P. Hopwood and D. H Crawford The role of EBV in post-transplant malignancies: a review J. Clin. Pathol., April 1, 2000; 53(4): 248 - 254. [Full Text] [PDF]

				Q. Tao, L. J. Swinnen, J. Yang, G. Srivastava, K. D. Robertson, and R. F. Ambinder Methylation Status of the Epstein-Barr Virus Major Latent Promoter C in Iatrogenic B Cell Lymphoproliferative Disease : Application of PCR-Based Analysis Am. J. Pathol., August 1, 1999; 155(2): 619 - 625. [Abstract] [Full Text] [PDF]