title: Quantitative multi-target RNA profiling in Epstein-Barr virus infected tumor cells.
abstract: Epstein-Barr virus (EBV) is etiologically linked to multiple acute, chronic and malignant diseases. Detection of EBV-RNA transcripts in tissues or biofluids besides EBV-DNA can help in diagnosing EBV related syndromes. Sensitive EBV transcription profiling yields new insights on its pathogenic role and may be useful for monitoring virus targeted therapy. Here we describe a multi-gene quantitative RT-PCR profiling method that simultaneously detects a broad spectrum (n=16) of crucial latent and lytic EBV transcripts. These transcripts include (but are not restricted to), EBNA1, EBNA2, LMP1, LMP2, BARTs, EBER1, BARF1 and ZEBRA, Rta, BGLF4 (PK), BXLF1 (TK) and BFRF3 (VCAp18) all of which have been implicated in EBV-driven oncogenesis and viral replication. With this method we determine the amount of RNA copies per infected (tumor) cell in bulk populations of various origin. While we confirm the expected RNA profiles within classic EBV latency programs, this sensitive quantitative approach revealed the presence of rare cells undergoing lytic replication. Inducing lytic replication in EBV tumor cells supports apoptosis and is considered as therapeutic approach to treat EBV-driven malignancies. This sensitive multi-primed quantitative RT-PCR approach can provide broader understanding of transcriptional activity in latent and lytic EBV infection and is suitable for monitoring virus-specific therapy responses in patients with EBV associated cancers.
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands.
authors.forename: A E
authors.initials: AE
authors.lastname: Greijer
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands.
authors.forename: O
authors.initials: O
authors.lastname: Ramayanti
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands.
authors.forename: S A W M
authors.initials: SA
authors.lastname: Verkuijlen
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands.
authors.forename: Z
authors.initials: Z
authors.lastname: Novalić
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: Juwana
authors.affiliation: Department of Pathology, VU University Medical Center and Cancer Center Amsterdam, The Netherlands. Electronic address: j.middeldorp@vumc.nl.
authors.forename: J M
authors.initials: JM
authors.lastname: Middeldorp
mesh_terms: D015415:Biomarkers; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000502:physiology; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics; D017735:Virus Latency / Q000235:genetics; D014779:Virus Replication
chemical_list: D015415:Biomarkers; D019309:Epstein-Barr Virus Nuclear Antigens; C099537:Epstein-Barr virus encoded RNA 1; D012333:RNA, Messenger; D012367:RNA, Viral; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C417029:EBV-encoded nuclear antigen 1
keywords: Epstein-Barr virus; Quantitative RT-PCR; RNA expression profile