RESUMO
The role of human adenovirus (HAdV) infection in different acute diseases, such as febrile exudative tonsillitis, conjunctivitis, and pharyngoconjunctival fever is well established. However, the relationships, if any, of HAdV persistence and reactivation in the development of the chronic adenotonsillar disease is not fully understood. The present paper reports a 3-year cross-sectional hospital-based study aimed at detecting and quantifying HAdV DNA and mRNA of the HAdV hexon gene in adenoid and palatine tonsil tissues and nasopharyngeal secretions (NPS) from patients with adenotonsillar hypertrophy or recurrent adenotonsillitis. HAdV C, B, and E were detectable in nearly 50% of the patients, with no association with the severity of airway obstruction, nor with the presence of recurrent tonsillitis, sleep apnea or otitis media with effusion (OME). Despite the higher rates of respiratory viral coinfections in patients with HAdV, the presence of other viruses, including DNA and RNA viruses, had no association with HAdV replication or shedding in secretions. Higher HAdV loads in adenoids showed a significant positive correlation with the presence of sleep apnea and the absence of OME. Although this study indicates that a significant proportion (~85%) of individuals with chronic adenotonsillar diseases have persistent nonproductive HAdV infection, including those by HAdV C, B, and E, epithelial and subepithelial cells in tonsils seem to be critical for HAdV C production and shedding in NPS in some patients, since viral antigen was detected in these regions by immunohistochemistry in four patients, all of which were also positive for HAdV mRNA detection.
Assuntos
Tonsila Faríngea/virologia , Infecções por Adenovirus Humanos/virologia , Tonsila Palatina/virologia , Replicação Viral , Tonsila Faríngea/patologia , Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Adenovírus Humanos/fisiologia , Adolescente , Criança , Pré-Escolar , Estudos Transversais , DNA Viral/isolamento & purificação , Feminino , Humanos , Hipertrofia , Lactente , Masculino , Tonsila Palatina/patologia , Tonsilite/virologiaRESUMO
OBJECTIVES: To evaluate the presence of viruses and bacteria in middle ear and adenoids of patients with and without otitis media with effusion (OME). METHODS: Adenoid samples and middle ear washes (MEW) were obtained from children with OME associated with adenoid hypertrophy undergoing adenoidectomy and tympanostomy, and compared to those obtained from patients undergoing cochlear implant surgery, as a control group. Specific DNA or RNA of 9 respiratory viruses (rhinovirus, influenza virus, picornavirus, syncytial respiratory virus, metapneumovirus, coronavirus, enterovirus, adenovirus and bocavirus) and 5 bacteria (S. pneumoniae, H. influenzae, M. catarrhalis, P. aeruginosa and S. aureus) were extracted and quantified by real-time PCR. RESULTS: 37 OME and 14 cochlear implant children were included in the study. At the adenoid, virus and bacteria were similarly detected in both OME and control patients. At the middle ear washes, however, a higher prevalence of bacteria was observed in patients with OME (p = 0.01). S. pneumoniae (p = 0.01) and M. catarrhalis (p = 0.022) were the bacteria responsible for this difference. Although total virus detection was not statistically different from controls at the middle ear washes (p = 0.065), adenovirus was detected in higher proportions in adenoid samples of OME patients than controls (p = 0.019). CONCLUSIONS: Despite both OME and control patients presented similar rates of viruses and bacteria at the adenoid, children with OME presented higher prevalence of S. pneumonia, M. catarrhalis in middle ear and adenovirus in adenoids when compared to controls. These findings could suggest that these pathogens could contribute to the fluid persistence in the middle ear.
Assuntos
Tonsila Faríngea/microbiologia , Tonsila Faríngea/virologia , Orelha Média/microbiologia , Orelha Média/virologia , Otite Média com Derrame/microbiologia , Otite Média com Derrame/virologia , Tonsila Faríngea/patologia , Bactérias/isolamento & purificação , Criança , Pré-Escolar , Implantes Cocleares , Orelha Média/patologia , Feminino , Humanos , Hipertrofia , Masculino , Otite Média com Derrame/patologia , Vírus/isolamento & purificaçãoRESUMO
Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR) in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (pâ=â0.05), and in the particular case of human enteroviruses, they were detected more frequently (pâ=â0.05) in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.
Assuntos
Tonsila Faríngea/virologia , Tonsila Palatina/virologia , Tonsilite/virologia , Viroses/virologia , Tonsila Faríngea/patologia , Adenovírus Humanos/genética , Adolescente , Criança , Pré-Escolar , Doença Crônica , DNA Viral/genética , Feminino , Bocavirus Humano/genética , Humanos , Lactente , Masculino , Tonsila Palatina/patologia , Vírus de RNA/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Tonsilite/epidemiologia , Tonsilite/genética , Tonsilite/patologia , Viroses/epidemiologia , Viroses/genética , Viroses/patologiaRESUMO
OBJECTIVE: Human herpesvirus 8 (HHV-8) and Epstein-Barr virus (EBV) are human pathogens associated to a number of neoplasms, including tumors of the Waldeyer's ring. Both viruses have been previously detected by in situ methods in tonsils and adenoids from children. HHV-8 was found in 6.8% of the cases and EBV in about one third of the cases. As they belong to the same gamma-herpesvirus subfamily and share some biological characteristics, it is of medical interest to further explore their possible relationship in the Waldeyer's ring, an issue not yet addressed in the specialized literature. The purpose of the present study is to compare the presence of EBV by in situ hybridization (ISH) in tonsils and adenoids from children up to 14 years of age in cases previously shown to be positive and negative for HHV-8. METHODS: Paraffin wax-embedded sections consisting of 38 tonsils and two adenoids from 40 patients were analyzed. HHV-8 was detected by ISH, using the T1-1 probe for the viral mRNA. EBV was also detected by ISH, using the EBER probe. Both probes and the detection systems were provided by Novocastra. RESULTS: HHV-8 was detected in 19 tonsils and one adenoid. The other 19 tonsils and one adenoid taken from the HHV-8-negative group were selected by pairing age and gender of patients with the HHV-8-positive group. In both groups EBV was detected in 13 cases and was negative in other 7. CONCLUSION: Although both viruses are related in many aspects, some biological and epidemiological features differ. This is reflected in the present results, as EBV is similarly detected in the groups negative and positive for HHV-8, favoring different mechanisms of spread.
Assuntos
Tonsila Faríngea/virologia , Herpesvirus Humano 4/isolamento & purificação , Herpesvirus Humano 8/isolamento & purificação , Tonsila Palatina/virologia , Tonsila Faríngea/química , Adolescente , Criança , Pré-Escolar , DNA Viral/análise , Feminino , Humanos , Hibridização In Situ , Lactente , Masculino , Tonsila Palatina/química , Inclusão em ParafinaRESUMO
OBJECTIVE: Human herpesvirus 8 (HHV8) has been associated with multicentric Castleman's disease, Kaposi's sarcoma and effusion non-Hodgkin's lymphoma. Epidemiological studies have shown seropositivity in variable proportions of populations. It seems to be sexually transmitted among adults and through oral contact among children. The virus has been demonstrated in desquamating oral epithelial cells, but there is no report on its presence in the Waldeyer's ring. The purpose of the present study is to detect HHV8 in tonsils and adenoids from children up to 20 years of age in which these organs had been surgically removed due to hypertrophy, using immunohistochemistry and in situ hybridization. METHODS: Paraffin wax-embedded sections consisting of 181 tonsils and 162 adenoids from 293 patients were analyzed. HHV8 was detected by immunohistochemistry (IHC) using the anti-LNA1 antibody (Novocastra) and the LSAB+ detection system (Dako). For the in situ hybridization (ISH), the T1-1 probe for the viral mRNA and the detection system used were provided by Novocastra. RESULTS: In 20 cases (6.83%), HHV8 was detected in cells morphologically characterized as lymphoid. In three of them epithelial cells were also positive. In 19 cases, the virus was detected in tonsils and in just 1 case in an adenoid. In all 20 cases detection was possible by ISH, whereas in only 2 of them there was a concomitant positivity by IHC. CONCLUSION: Our data support the oral route of contamination by HHV8 in children, in whom tonsils and adenoids may harbor the virus. It is found especially in tonsils and only rarely in adenoids. In these organs, ISH is the method of choice to detect this virus, probably due to the small amount of viral proteins.
Assuntos
Tonsila Faríngea/virologia , Herpesvirus Humano 8/isolamento & purificação , Imuno-Histoquímica/métodos , Hibridização In Situ/métodos , Tonsila Palatina/virologia , Tonsila Faríngea/cirurgia , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Infecções por Herpesviridae/virologia , Humanos , Hipertrofia/cirurgia , Lactente , Masculino , Tonsila Palatina/cirurgia , Sarcoma de Kaposi/virologiaRESUMO
OBJECTIVE: Herpes simplex virus (HSV) has been described as cause of acute tonsillitis. It has also been found in nasopharyngeal florid lymphoid infiltrate, mostly composed of CD4+, CD56+ T-cells, simulating lymphoma. In spite of its widespread prevalence in latent form, to the best of our knowledge no study is available on in situ detection of HSV in chronically hyperplastic nasopharyngeal lymphoid tissue. The purpose of the present study was to search for the presence of HSV 1 and 2 in 21 adenoids and 15 tonsils from children (2-12 years of age) in which these organs had been surgically removed due to hypertrophy. METHODS: Paraffin wax-embedded sections from the 36 cases were submitted to the in situ hybridization technique, using the biotinilated probe to Herpes simplex virus 1 and 2 (Pan Path, Amsterdam) and the Rembrandt Universal DISH & HRP Detection Kit (Pan Path, Amsterdam). Positive control consisted of a previously tested Herpes infected lung. RESULTS: In none of the 36 cases studied were positive nuclei detected in adenoid and tonsils, either in lymphoid, in stroma or in epithelial cells, as those seen in the positive control. CONCLUSION: HSV does not seem to be implied in tonsil or adenoid chronic lymphoid hyperplasia. These organs do not seem to harbor the virus latently, or the amount of virus is too low to be detected without amplification methods.
Assuntos
Tonsila Faríngea/imunologia , Tonsila Faríngea/virologia , Antígenos CD4/imunologia , Antígeno CD56/imunologia , Herpes Simples/complicações , Herpes Simples/imunologia , Herpesvirus Humano 1/imunologia , Herpesvirus Humano 2/imunologia , Hibridização In Situ/métodos , Tonsilite/imunologia , Tonsilite/virologia , Tonsila Faríngea/patologia , Criança , Pré-Escolar , Feminino , Humanos , Hiperplasia/imunologia , Hiperplasia/patologia , Tecido Linfoide/imunologia , Tecido Linfoide/patologia , Masculino , Nasofaringe/imunologia , Nasofaringe/patologia , Tonsilite/patologiaRESUMO
Epstein-Barr virus (EBV) has been closely associated with undifferentiated nasopharyngeal carcinoma (NPC) and T/NK nasal non Hodgkin lymphoma. Nevertheless, the presence of EBV in non neoplastic lymphoid tissue of the nasopharynx has been rarely investigated. In a previous study by our group, using in situ hybridization to detect EBV in adenoids of children (2-13 years old) resected because of nasal obstruction due to hypertrophy, we found EBV genome in 72% of the cases. It was now intended to study the frequency of EBV expression in adenoids from children that underwent surgical removal, belonging to a lower age group (1-2 years old). It was also intended to establish which lymphoid subsets are involved in this infection. Adenoidal paraffin sections from 21 patients aged 1-2 years old (mean 1.6 years), 15 males and six females were submitted to double labeling: in situ hybridization with EBER 1/2 probes to detect EBV and immunohistochemistry to determine the lymphocyte typing of EBV-positive cells (CD20 for B-lymphocytes, CD3 for T-lymphocytes and CD56 and CD57 for NK-cells). Among 21 patients, seven showed positive lymphoid cells for EBV (33%). In almost all cases, EBV-positive cells were also CD20-positive. Some EBV-positive cells showed no labeling with any of the lymphoid markers, but in no instance they were positive for CD3, CD56 or CD57. This study confirms the preferential infection of B-lymphocytes by EBV, which in some instances can down regulate the expression of CD20.