Abstract
Our study aimed at investigating the distribution, persistence and interchange of viral strains among peripheral blood mononuclear cells (PBMC), plasma and saliva of primary Epstein-Barr virus (EBV) infection subjects. Twelve infectious mononucleosis (IM) patients and eight asymptomatic individuals (AS) with primary EBV infection were followed longitudinally at several time points for one year from the time of diagnosis, when blood and saliva samples were collected and separated into PBMC, plasma and saliva, representing circulating B cell, plasma and epithelial cell compartments, respectively. To survey the viral strains, genotyping assays for the natural polymorphisms in two latent EBV genes, EBNA2 and LMP1, were performed and consisted of real-time PCR on EBNA2 to distinguish type 1 and 2 viruses, fluorescent-based 30-bp typing assay on LMP1 to distinguish deletion and wild type LMP1, and fluorescent-based heteroduplex tracking assays on both EBNA2 and LMP1 to distinguish defined polymorphic variants. No discernible differences were observed between IM patients and AS. Multiple viral strains were acquired early at the start of infection. Stable persistence of dominant EBV strains in the same tissue compartment was observed throughout the longitudinal samples. LMP1-defined strains, China 1, China 2 and Mediterranean+, were the most common strains observed. EBNA2-defined groups 1 and 3e predominated the PBMC and saliva compartments. Concordance of EBNA2 and LMP1 strains between PBMC and saliva suggested ready interchange of viruses between circulating B cell and epithelial cell pools, whilst discordance of viral strains observed between plasma and PBMC/saliva indicated presence of viral pools in other undetermined tissue compartments. Taken together, the results indicated that the distribution, persistence and interchange of viral strains among the tissue compartments are more complex than those proposed by the current model of EBV life cycle.
Highlights
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus infecting more than 90% of the adult human populations
Typing assays and Heteroduplex tracking assay (HTA) were performed on 20 cases, which included 12 childhoodinfectious mononucleosis (IM) and 8 asymptomatic (AS) cases of primary EBV infection
Since there was no observable difference between the strain profiles of IM and asymptomatic individuals (AS) subjects, these 20 cases can be considered as one cohort of EBV primary infection for the purpose of this study
Summary
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus infecting more than 90% of the adult human populations. Two major types of EBV, namely type 1 and type 2 ( known as types A and B), are distinguished by sequence polymorphisms in Epstein-Barr nuclear antigen (EBNA)-2, -3A, -3B and -3C genes [1,2] They occur worldwide but with different geographical distribution. Further analysis of EBNA2 and Epstein-Barr encoded RNA (EBER) gene sequences of EBV genomes contained in clinical samples of different malignant and benign EBV-associated diseases identified common variant strains of EBV1, namely, groups 1, 2, 3a, 3b and 3e [6]. Another highly polymorphic EBV gene, namely LMP1, is frequently used in molecular epidemiological studies of EBV infection. A total of seven LMP1 strains, including the prototype B95–8 strain, were defined
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