Detection of the Epstein–Barr virus, Human Bocavirus and novel KI and KU polyomaviruses in adenotonsillar tissues

doi:10.1016/j.ijporl.2015.01.007

International Journal of Pediatric Otorhinolaryngology

Volume 79, Issue 3, March 2015, Pages 423-427

https://doi.org/10.1016/j.ijporl.2015.01.007 Get rights and content

Abstract

Objectives

We aimed to investigate the seasonal disturbations and the rates of detection of Epstein–Barr virus (EBV), Human Bocavirus (HBoV), and polyomaviruses KI and WU (KIPyV and WUPyV) in adenoid and tonsil tissues during the absence of acute infection symptoms.

Study design

Cross-sectional prospective study.

Setting

Tertiary hospital.

Methods

DNA expressions of EBV, HBoV, polyomaviruses KIPyV and WUPyV were investigated in children with chronic tonsillar and adenoidal diseases using real-time polymerase chain reaction. The patients were grouped as follows: adenoid, recurrent tonsillitis and hypertrophic tonsillitis group. The relationships of the expressions of these viruses with age, gender, recurrent infection and airway obstruction were also analyzed. Seasonal variations in rates of detection of these viruses in adenoid and tonsil tissues were also investigated.

Results

Considering adenoid specimens, HBoV was found to be the most frequent virus with the rate of 43.1%. In specimens of recurrent tonsillitis and hypertrophic tonsils, EBV was the most frequently encountered virus (53.8%, and 32.0%, respectively). In children with hypertrophic adenoids, while HBoV was detected to be positive throughout the year, EBV was present throughout the year in children with recurrent tonsillitis.

Conclusions

The detection of HBoV and EBV throughout the year in samples of children with asymptomatic chronic adenotonsillar diseases may indicate that these viruses may remain persistently in lymphoepithelial tissues of upper respiratory tract. Virus persistence may have a pathogenetic potential for development of lymphoid hypertrophy and a chronic stimulatory effect for inflammation.

Introduction

The palatine tonsils (tonsils) and pharyngeal tonsils (adenoids) are the most important structures in the Waldeyer's ring. Recurrent and chronic diseases of tonsils and adenoids are highly frequent in children and lead to chronic activation of the immune response, resulting in hypertrophy of the adenoidal and tonsillar lymphoid tissues [1]. These reactions can adversely affect a child, especially when there is hypertrophy and/or recurrent infection. This hypertrophy may lead to several other diseases and symptoms such as hyponasality, snoring, obstructive sleep apnea syndrome, acute otitis media, otitis media with effusion, middle ear atelectasis, cholesteatoma formation, slow feeding, acute sinusitis, abnormal facial development and behavioral problems when hypertrophy causes airway obstruction [2], [3], [4]. For these reasons, tonsillectomy with or without adenoidectomy is the most frequent surgical procedure performed by otorhinolaryngologists [5].

Yet, the etiologies of chronic adenotonsillar diseases are largely unknown. Generally, bacteria are considered to be the most likely causative agents in adenotonsillar diseases [6], [1].

However, many viruses are detected in tonsillar and adenoid tissues recently [1]. Despite these findings, systematic studies of viruses in adenoids and tonsils have been rare, especially in recurrent and/or hypertrophic adenotonsillar diseases.

In the present study, DNA expressions of Human Bocavirus (HBoV), Epstein–Barr virus (EBV) and polyomaviruses KI and WU (KIPyV and WUPyV) were investigated in children with tonsillar and adenoidal diseases using real-time polymerase chain reaction (RT-PCR). The relationships of expressions of these viruses with other clinical characteristics of children, such as patient age, gender, recurrent infection and airway obstruction were also analyzed. Seasonal variations in rates of detection of these viruses in adenoid and tonsil tissues during the absence of acute infection symptoms were also investigated. The results of this study provide a new insight about the roles of these viruses in the pathogenesis of recurrent tonsillitis and tonsillar/adenoidal hypertrophy.

Section snippets

Patients and specimen collections

This cross-sectional study was carried out from October 2012 to August 2014. A total of 51 children (31 boys and 20 girls) aged 2 to 15 (mean 6.18 ± 2.83) years were enrolled in the study. All children had adenoidal hypertrophy and tonsillar hypertrophy/or recurrent tonsillitis with indications for adenotonsillectomy. Exclusion criteria were: acute respiratory infection at the time of surgery, antibiotic treatment within 4 weeks prior to surgery, immunological disorders, and chronic medical

Viral detection

A total of 51 adenoid samples and 51 tonsil samples were tested to determine the sensitivity of the RT-PCR assays. The demographic characteristics of the patients were shown in Table 1. Considering adenoid specimens, EBV was found positive in 41.2%, Bocavirus in 43.1%, WUPyV in 7.8% and KIPyV was positive in none. While the incidence rates were statistically similar for EBV and HBoV in adenoid samples (p = 0.841), WUPyV was found to be positive significantly less frequently than EBV and Bocavirus

Discussion

In various studies, EBV has been identified as one of the most frequently met viral agents which cause recurrent tonsillitis [7], [8], [9], [10]. Tonsils are also suggested to be the onset and replication site for the infections [8]. In our study, we found out that EBV was positive most frequently in tonsillar samples of children with recurrent tonsillitis (53.8%) using real-time PCR. This was followed by samples of children with adenoid hypertrophy (41.2%) and tonsillar hypertrophy (32.0%).

Conflict of interest statement

The authors have no conflicts of interest to disclose and have no financial disclosures to make.

Acknowledgment

The authors thank ADÜ-BAP (Adnan Menderes Üniversitesi Bilimse Araştırma Projeleri) for financial support.

References (22)

C.Y. Chien et al.
The clinical significance of adenoid–choanae area ratio in children with adenoid hypertrophy
Int. J. Pediatr. Otorhinolaryngol.
(2005)
T.K. Erdag et al.
Pathologic evaluation of routine tonsillectomy and adenoidectomy specimens in the pediatric population: is it really necessary?
Int. J. Pediatr. Otorhinolaryngol.
(2005)
L.H. Endo et al.
Detection of Epstein–Barr virus in tonsillar tissue of children and the relationship with recurrent tonsillitis
Int. J. Pediatr. Otorhinolaryngol.
(2001)
E.P. Dias et al.
Detection of Epstein–Barr virus in recurrent tonsillitis
Braz. J. Otorhinolaryngol.
(2009)
T. Dalianis et al.
Human polyomaviruses in disease and cancer
Virology
(2013)
L. Ren et al.
WU and KI polyomavirus present in the respiratory tract of children, but not in immunocompetent adults
J. Clin. Virol.
(2008)
M. Bergallo et al.
Real time PCR TaqMan assays for detection of polyomaviruses KIV and WUV in clinical samples
J. Virol. Methods
(2009)
M. Babakir-Mina et al.
Identification of the novel KI and WU polyomaviruses in human tonsils
J. Clin. Virol.
(2009)
X.C. Xue et al.
Prevalence of human papillomavirus and Epstein–Barr virus DNA in Chinese children with tonsillar and/or adenoidal hypertrophy
J. Med. Virol.
(2014)
J.M. Bernstein
Waldeyer's ring and otitis media: the nasopharyngeal tonsil and otitis media
Int. J. Pediatr. Otorhinolaryngol.
(1999)

G. Papaioannou et al.

Age-dependent changes in the size of adenotonsillar tissue in childhood: implications for sleep-disordered breathing

J. Pediatr.

(2013)

Cited by (32)

Quantitative analysis of respiratory viral distribution in forensic autopsy cases
2022, Forensic Science International
Citation Excerpt :
According to previous studies, respiratory viruses were detected in the surgically removed tonsils of children with chronic adenotonsillar diseases, who did not exhibit infectious symptoms. Those studies indicated that certain respiratory viruses could persist in tonsils after the acute phase of infection [37,38]. Thus, viruses might be detectable in the spleen and the tonsils after an acute infection since they are both lymphatic organs.
Respiratory viruses can cause fatal systemic infections; therefore, post-mortem diagnosis is essential in forensic autopsy cases. However, little is known regarding the distribution of respiratory viruses in the body. In this study, we investigated the anatomical distribution of respiratory viruses in 48 forensic autopsy cases suspected of viral infections at our institute. Fast Track Diagnostics (FTD) Respiratory Pathogens 21 was used as a screening test for 20 respiratory viruses in nasopharyngeal swabs. In cases with positive results for virus detection by the screening test, the detected viruses were quantified in body fluid and organ specimens by virus-specific real-time reverse transcription polymerase chain reaction (RT-PCR) and digital PCR. Viruses were detected in 33 cases, with the viral distribution and load differing among the cases. Since various respiratory viruses were detected from the nasopharyngeal swab and its viral load was higher than those of other body fluid specimens, the nasopharyngeal swab was suggested as a useful specimen for the post-mortem detection of respiratory viruses. Viruses were detected in almost all specimens including the serum in six cases. Considering the viral distribution in the body, pathological findings, and ante-mortem symptoms, these cases were presumed to be systemically infected, having died in the acute infection phase. In conclusion, the anatomical distribution of respiratory viruses can help indicate ante-mortem systemic conditions and the cause of death.
Surgery for adenotonsillar hypertrophy and otitis media in children is less demanded in quarantine times
2022, International Journal of Pediatric Otorhinolaryngology
Citation Excerpt :
Seasonality of viral respiratory infections in general is a universally acknowledged phenomenon, and within the regions of the temperate climate zone, such as Greece, the peak occurs in winter [19]. Nevertheless, aside from the acute manifestations of an URTI, viruses remain indefinitely in the adenotonsillar tissues [12]. Hence, a combined proliferative effect of respiratory viruses on lymphoepithelial tissues could be postulated: chronic antigenic stimulation during the carrier state, upon which superimposed exacerbations in the form of new infections accumulate.
In the COVID-19 era, extreme measures of social distancing have contained the spread of common viral respiratory infections, which are involved in the pathogenesis of Adenotonsillar Hypertrophy (ATH), and Chronic Otitis Media with Effusion (COME), the two commonest chronic ENT diseases of childhood. This study examines the lockdown effect on the number of procedures performed for these two conditions.
The consecutive charts of 650 and 195 children being operated respectively for ATH and COME during the quarantine (05/2020-02/2021) and unrestrained (05/2019-02/2020) periods were retrospectively reviewed. Surgical treatment of ankyloglossia, performed in 103 patients during the same periods was employed as a control procedure.
Adenotonsillectomies and tympanostomies significantly decreased in the lockdown phase by 52% (P < 0.001) and 74% (P < 0.001), respectively, whereas control procedure counts increased by 25%. In terms of seasonal variation, ATH-related surgeries were significantly reduced during the winter season of the pandemic by 73% (P < 0.001), in comparison with the corresponding months of the unrestrained period. School-aged children received significantly fewer operations for ATH (−59%) than preschoolers (−42%), as a result of the lockdown (P = 0.044).
When the child's exposure to respiratory pathogens is minimal, as in the case of lockdown, a noticeable decline occurs in the incidence of ATH and COME indicated for surgical treatment. Chronic low-grade inflammation, boosted by repetitive viral infections seems to underlie both conditions. Timely, effective isolation measures might reverse the disease process and keep the child away from the Operating Room.
Human bocaviruses and paediatric infections
2019, The Lancet Child and Adolescent Health
Citation Excerpt :
In one study, whole parvovirus particles were detected in cerebral spinal fluid by electron microscopy.18 HBoV1 DNA has been detected in tissue from hypertrophic tonsils or adenoids in children.23,71,72 The pathogenic role of the viruses in these tissues remains uncertain.
Human bocavirus 1 (HBoV1), belonging to the Parvoviridae family, was discovered in 2005, in nasopharyngeal samples from children with respiratory tract infections. Three additional bocaviruses, HBoV2–4, were discovered in 2009–10. These viruses have mainly been found in faecal samples and their role in human diseases is still uncertain. HBoV1 causes a wide spectrum of respiratory diseases in children, including common cold, acute otitis media, pneumonia, bronchiolitis, and asthma exacerbations. HBoV1 DNA can persist in airway secretions for months after an acute infection. Consequently, acute HBoV1 infection cannot be diagnosed with standard DNA PCR; quantitative PCR and serology are better diagnostic approaches. Because of their high clinical specificity, diagnostic developments such as HBoV1 mRNA and antigen detection have shown promising results. This Review summarises the knowledge on human bocaviruses, with a special focus on HBoV1.
Could human bocavirus be a causative agent of parotitis in children?
2019, Enfermedades Infecciosas y Microbiologia Clinica
Oncogenic DNA viruses found in salivary gland tumors
2017, Oral Oncology
Previous investigations studying the association of DNA viruses with salivary gland tumors (SGTs) have led to conflicting results. The aim of this study was to determine the prevalence of different DNA viruses by using a highly sensitive assay in a multi-center series of over 100 fresh frozen salivary gland samples.
DNA was isolated from 84 SGTs (80 parotid tumors and 4 submandibular gland tumors) and 28 normal salivary tissue samples from 85 patients in Northeast Italy. Using a highly sensitive type-specific multiplex genotyping assay, we analyzed the samples for the presence of DNA from 62 different viruses including 47 papillomaviruses, 10 polyomaviruses, and 5 herpesviruses.
We observed a high prevalence of beta human papillomavirus DNA in malignant tumors. In contrast, polyomavirus DNA was present in benign, malignant, and non-tumor control samples. Most striking was the significant distribution of herpesvirus DNA in the SGT samples, in particular the high prevalence of Epstein-Barr type 1 and type 2 DNA in Warthin’s tumor samples.
Our data provides evidence for the presence of DNA viruses in SGTs. Mechanistic studies are needed to further attribute tumor formation to these viruses.
Human bocavirus 1 infection of CACO-2 cell line cultures
2017, Virology
Citation Excerpt :
This might be an outcome of different and possibly overlapping mechanisms of virus-cell interactions, including mutations in the progeny genome that have a profound effect in the viral protein functions, namely the virus entry and genome replication. Overall, these results can provide insights into the observations that HBoV1 can be detected in human tissues other than the respiratory epithelium (Gunel et al., 2015; Norja et al., 2012; Schildgen et al., 2013). In Caco-2 cell cultures infected with HBoV1 and supplemented with DEAE-dextran (1 µg/ml) at 33 °C and 37 °C, the presence of HBoV1 was observed in the supernatants and the cells attached to the substrate from 1 to 6 dpi, in contrast to cultures maintained with standard EMEM (Table 1 and Fig. 3A and B).
Human bocavirus 1 (HBoV1) is a parvovirus associated with pneumonia in infants. It has been detected in different tissues, including colorectal tumors. In this study, we investigated whether Caco-2 cell line, derived from human colon cancer, can be utilized as a model for HBoV1 replication. We demonstrate HBoV1 replication in Caco-2 cultures supplemented with DEAE-dextran after inoculation with respiratory material from infected patients presenting with acute respiratory infection. A viral cycle of rapid development is displayed. However, in spite of HBoV1 DNA 4-fold increment in the supernatants and monolayers by day 1, evidencing that the system allows the virus genome replication after the entry occurred, infectious progeny particles were not produced. These results are consistent with an infection that is limited to a single growth cycle, which can be associated to mutations in the NS1 and VP1/VP2 regions of HBoV1 genome. Further research will contribute to fully elucidate these observations.

View all citing articles on Scopus

View full text

Detection of the Epstein–Barr virus, Human Bocavirus and novel KI and KU polyomaviruses in adenotonsillar tissues

Abstract

Objectives

Study design

Setting

Methods

Results

Conclusions

Introduction

Section snippets

Patients and specimen collections

Viral detection

Discussion

Conflict of interest statement

Acknowledgment

Int. J. Pediatr. Otorhinolaryngol.

Int. J. Pediatr. Otorhinolaryngol.

Int. J. Pediatr. Otorhinolaryngol.

Braz. J. Otorhinolaryngol.

Virology

J. Clin. Virol.

J. Virol. Methods

J. Clin. Virol.

Prevalence of human papillomavirus and Epstein–Barr virus DNA in Chinese children with tonsillar and/or adenoidal hypertrophy

J. Med. Virol.

Waldeyer's ring and otitis media: the nasopharyngeal tonsil and otitis media

Int. J. Pediatr. Otorhinolaryngol.

Age-dependent changes in the size of adenotonsillar tissue in childhood: implications for sleep-disordered breathing

J. Pediatr.