RESUMEN

Water buffalo (Bubalus bubalis) farming is increasing in many regions of the world due to the species' ability to thrive in environments where bovine cattle would struggle. Despite water buffaloes being known for their resistance to diseases, there is a lack of data about the diversity of the microbiome of the species. In this study, we examined the virome diversity in palatine tonsils collected from animals from the island of Marajó, northern Pará state, Brazil, which harbors the largest bubaline flock in the country. Tonsil fragments from 60 clinically healthy bubalines were randomly selected from a sample of 293 animals. The samples were purified, extracted, and randomly amplified with phi29 DNA polymerase. After amplification, the products were purified and sequenced. Circular DNA viruses were predominant in the tonsils' virome. Sequences of genome segments representative of members of the genera Alphapolyomavirus (including a previously unreported bubaline polyomavirus genome) and Gemycircularvirus were identified, along with other not yet classified circular virus genomes. As the animals were clinically healthy at the time of sampling, such viruses likely constitute part of the normal tonsillar virome of water buffaloes inhabiting the Ilha do Marajó biome.

Asunto(s)

Búfalos , Tonsila Palatina , Filogenia , Poliomavirus , Animales , Búfalos/virología , Tonsila Palatina/virología , Brasil , Poliomavirus/genética , Poliomavirus/aislamiento & purificación , Poliomavirus/clasificación , Viroma , ADN Viral/genética , Genoma Viral

RESUMEN

Herpesviruses are significant pathogens of ruminants. In water buffaloes (Bubalus bubalis), however, herpesviruses have not been thoroughly studied. Although bubaline alphaherpesvirus 1 (BuAHV1) and bovine alphaherpesvirus 1 (BoAHV1) have already been recovered from water buffaloes, to date, no reports on the occurrence of bovine alphaherpesvirus 5 (BoAHV5) in these animals have been published. Therefore, the aim of this study was to search for BuAHV1, BoAHV1, and BoAHV5 in palatine tonsils of apparently healthy water buffaloes from the Pará state, Northern Brazil. Tissue samples of tonsils (n = 293) were screened by a nested PCR (nPCR) targeting a region of UL44 (gC coding gene), followed by sequencing, to detect and differentiate between the viral types. Viral genome segments were detected in 18 out of 293 (6.1%) of the palatine tonsil samples. Two animals carried genomes of BoAHV1 only, eleven animals carried BoAHV5 genomes only, and four animals carried BuAHV1 only. Another animal had both BoAHV1 and BoAHV5 genomes in its tonsils. No infectious virus could be recovered from any of the samples. The BuAHV1 sequences identified here were more closely related to BuAHV1 genomes identified in India. Phylogenetic analyses suggested a closer relationship between the recovered BoAHV5 and BuAHV1 genomes. Therefore, evidence is provided here to confirm that not only BoAHV1 and BuAHV1, but also BoAHV5, can infect water buffaloes. This report highlights (i) the first detection of BoAHV5 in water buffaloes and (ii) the occurrence of coinfections with BoAHV1 and BoAHV5 in that species. Such findings and the similarity of BoAHV5 to Indian herpesvirus genomes suggest that the origin of type 5 may be linked to recombinations between bovine and bubaline herpesviruses within bubalines, since the scenario for generation of recombinants in buffaloes is potentially present.

Asunto(s)

Alphaherpesvirinae , Búfalos , Infecciones por Herpesviridae , Tonsila Palatina , Animales , Bovinos , Alphaherpesvirinae/genética , Alphaherpesvirinae/aislamiento & purificación , Alphaherpesvirinae/clasificación , Brasil , ADN Viral/genética , Genoma Viral , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/virología , Tonsila Palatina/virología , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN

RESUMEN

BACKGROUND: In developing countries, Epstein-Barr virus (EBV) infection is mostly asymptomatic in early childhood. EBV persistence may lead to different malignancies, such as B cell derived lymphomas. In Argentina, most children are seropositive at three years and an increased association between EBV and lymphoma was proved in children under 10 years old by our group. OBJECTIVE: Our aim was to characterize EBV infection at the site of entry and reactivation of viral infection -the tonsils- in order to better understand the mechanism of viral persistence in pediatric patients. METHODS: A cohort of 54 patients was described. We assessed specific antibodies profiles in sera; viral proteins presence by IHC on FFPE samples and EBV type from fresh tissue. RESULTS: EBV type 1 was prevalent, mostly in the youngest patients. Asymptomatic primary infected patients presented higher viral loads and Latency 0/I or II patterns, whereas the Latency III pattern was observed mostly in healthy carriers. There were no differences between groups in the expression of viral lytic antigens. This study discloses new features in patients undergoing primary infection from a developing population. Low viral inoculum and restricted viral antigen expression may be responsible for the lack of symptoms in children from our country.

Asunto(s)

Infecciones por Virus de Epstein-Barr/virología , Adolescente , Anticuerpos Antivirales/sangre , Antígenos Virales/metabolismo , Argentina , Niño , Preescolar , Países en Desarrollo , Femenino , Herpesvirus Humano 4/inmunología , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 4/metabolismo , Humanos , Lactante , Masculino , Tonsila Palatina/virología , Carga Viral , Proteínas Virales/metabolismo , Latencia del Virus

RESUMEN

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.

Asunto(s)

Tonsila Faríngea/virología , Infecciones por Adenovirus Humanos/virología , Tonsila Palatina/virología , Replicación Viral , Tonsila Faríngea/patología , Infecciones por Adenovirus Humanos/diagnóstico , Adenovirus Humanos/clasificación , Adenovirus Humanos/aislamiento & purificación , Adenovirus Humanos/fisiología , Adolescente , Niño , Preescolar , Estudios Transversales , ADN Viral/aislamiento & purificación , Femenino , Humanos , Hipertrofia , Lactante , Masculino , Tonsila Palatina/patología , Tonsilitis/virología

RESUMEN

INTRODUCTION: Epstein-Barr Virus (EBV) infection prevails in underdeveloped and developing countries. The tonsils seem to be candidate replication sites for EBV and some studies have exposed a close association among viral infections and chronic tonsillitis. The objective of this study was identifying the EBV prevalence in Mexican patients who had undergone tonsillectomy because of chronic tonsillitis. METHODOLOGY: Frozen tissues and medical records were obtained from 50 Mexican patients. DNA was extracted and subjected to PCR to amplify the EBER-2 region of EBV. Next, the patients were classified according to general and clinical characteristics searching a relation with the EBV-DNA positivity. RESULTS: EBV genome was detected in 46% (23/50) of the analysed tonsil tissues. Trends were found regarding the relationship of viral presence with lower values in terms of age (6.1 ± 2.8 vs 7.6 ± 3.7) , a greater degree of hypertrophy (3.5 ± 0.4 vs 3.0 ± 0.6) and an increase in the number of episodes of tonsillitis (11 ± 7.4 vs 9 ± 6.5). CONCLUSIONS: The prevalence found of EBV-DNA positivity in tonsillar tissues from patients diagnosed with chronic tonsillitis , supports the fact that palatine tonsils can be occupied by EBV and highlights the importance of conducting future studies focused on understanding the role of the EBV infection in chronic inflammatory processes in the population involved in this study.

Asunto(s)

ADN Viral/análisis , Infecciones por Virus de Epstein-Barr/epidemiología , Infecciones por Virus de Epstein-Barr/virología , Herpesvirus Humano 4/aislamiento & purificación , Tonsila Palatina/virología , Tonsilitis/epidemiología , Tonsilitis/virología , Adolescente , Niño , Preescolar , Enfermedad Crónica , ADN Viral/genética , Femenino , Herpesvirus Humano 4/genética , Humanos , Lactante , Recién Nacido , Masculino , México/epidemiología , Reacción en Cadena de la Polimerasa , Prevalencia

RESUMEN

Epstein Barr virus (EBV) gains access to the host through tonsillar crypts. Our aim was to characterize microenvironment composition around EBV+ cells in tonsils from pediatric carriers, to disclose its role on viral pathogenesis. LMP1 expression, assessed by immunohistochemistry (IHC), was used to discriminate EBV + and - zones in 41 tonsil biopsies. Three regions were defined: Subepithelial (SE), interfollicular (IF) and germinal center (GC). CD8, GrB, CD68, IL10, Foxp3, PD1, CD56 and CD4 markers were evaluated by IHC; positive cells/100 total cells were counted. CD8+, GrB+, CD68+ and IL10+ cells were prevalent in EBV+ zones at the SE region (p < 0.0001, p = 0.03, p = 0.002 and p = 0.002 respectively, Wilcoxon test). CD4+ and CD68+ cell count were higher in EBV + GC (p = 0.01 and p = 0.0002 respectively, Wilcoxon test). Increment of CD8, GrB and CD68 at the SE region could indicate a specific response that may be due to local homing at viral entry, which could be counterbalanced by IL10, an immunosuppressive cytokine. Additionally, it could be hypothesized that CD4 augment at the GC may be involved in the EBV-induced B-cell growth control at this region, in which macrophages could also participate.

Asunto(s)

Infecciones por Virus de Epstein-Barr/inmunología , Herpesvirus Humano 4/inmunología , Inmunidad Celular , Tonsila Palatina/patología , Tonsila Palatina/virología , Adolescente , Biomarcadores/análisis , Niño , Preescolar , Estudios de Cohortes , Femenino , Humanos , Masculino , Proteínas de la Matriz Viral/análisis

RESUMEN

INTRODUCTION: Porcine cytomegalovirus (PCMV) causes rhinitis in both young and older pigs. The present study describes the detection and characterization of shedding profiles of PCMV in nine farrow-to-finish Brazilian swine herds. METHODOLOGY: Tonsil swabs from sows, nursery and grow-finish pigs of nine farrow-to-finish commercial herds (n = 756) were evaluated for the presence of PCMV by PCR. RESULTS: The virus was detected in all herds. Positive samples were concentrated in piglets of ages varying from 40 to 60 days (nursery phase), while none of the sows were positive for PCMV detection. CONCLUSIONS: These findings corroborate the literature regarding PCMV worldwide distribution, and introduce the first report of PCMV shedding profile in Brazilian pig farms.

Asunto(s)

Infecciones por Citomegalovirus/veterinaria , Citomegalovirus/aislamiento & purificación , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/virología , Esparcimiento de Virus , Animales , Brasil/epidemiología , Infecciones por Citomegalovirus/epidemiología , Infecciones por Citomegalovirus/virología , Tonsila Palatina/virología , Porcinos

RESUMEN

Epstein-Barr virus (EBV) is a B lymphotropic human herpesvirus. Two models, germinal center (GC) and direct infection, describe how EBV infects B-cells. Since in Argentina primary infection is mostly subclinical at young ages, children represent an interesting population where to analyze EBV infection, especially considering that most studies are usually performed in adults. Tonsil biopsies from pediatric carriers were studied to describe infection characteristics. EBV+ lymphocytes at the interfollicular region were mainly observed. Latency III pattern in subepithelial (SubEp) lymphocytes was observed at young ages, probably indicating a recent infection. In older patients EBV was mostly detected in epithelial cells, suggesting that they could have been infected some time ago. This finding was sustained by tonsillar viral load, which was higher in cases with LMP1+SubEp cells vs. LMP1+nonSubEp cells (p = 0.0237, Mann-Whiney test). Latency III was prevalent and related to the GC, while latency II was associated with non-GC (p = 0.0159, χ2 test). EBERs+/IgD+ cells were statistically prevalent over EBERs+/CD27+ cells (p = 0.0021, χ2 test). These findings indicated that both EBV infection models are not mutually exclusive and provide some basis for further understanding of EBV infection dynamics. Moreover, we provide a more accurate explanation of EBV infection in pediatric asymptomatic carriers from a developing country.

Asunto(s)

Linfocitos B/virología , Células Epiteliales/virología , Infecciones por Virus de Epstein-Barr/virología , Antígenos Nucleares del Virus de Epstein-Barr/metabolismo , Herpesvirus Humano 4/fisiología , Tonsila Palatina/citología , Adolescente , Adulto , Argentina , Niño , Preescolar , Países en Desarrollo , Antígenos Nucleares del Virus de Epstein-Barr/genética , Femenino , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/inmunología , Humanos , Masculino , Modelos Teóricos , Tonsila Palatina/virología , Estudios Retrospectivos , Carga Viral , Latencia del Virus

RESUMEN

The role of tonsils in oral immunity has been described. However, the pathogenesis of HIV infection in these organs is still unclear. The aim of this study is to perform histological and immunohistochemical analysis of the palatine and lingual tonsils of autopsied individuals with or without HIV infection. Twenty-six autopsied individuals with HIV infection (HI) (n=13) and without HIV infection (CO) (n=13) were selected. Palatine and lingual tonsil fragments were collected for histological and immunohistochemical analysis. We found in the HI group a higher frequency of hyaline degeneration in both palatine and lingual tonsils; smaller follicle areas, and a higher percentage of collagen in comparison with the CO group. In the HI group, there was higher density of blood vessels in palatine tonsils than in the CO group. In the HI group, there were significant positive correlations between palatine and lingual tonsils and the area of lymphoid follicles, and between the percentage of blood vessels and collagen in palatine tonsils. In addition, there was a significant negative correlation between the percentage of collagen and lymphoid follicle area in both palatine and lingual tonsils in the HI group. These findings suggest that the immune functions of these tonsils are prejudiced by fibrosis. Therapies to reduce the neoformation of collagen are required to improve immune function of organs against pathogens.

Asunto(s)

Infecciones por VIH/inmunología , Ganglios Linfáticos/inmunología , Tonsila Palatina/inmunología , Lengua/patología , Adulto , Femenino , Fibrosis , Infecciones por VIH/patología , Humanos , Mucosa Intestinal/inmunología , Mucosa Intestinal/virología , Ganglios Linfáticos/virología , Masculino , Persona de Mediana Edad , Tonsila Palatina/virología , Lengua/inmunología

RESUMEN

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) can both establish latency in the trigeminal ganglion. Non-neural sites of latency have been described for BoHV-1 but not for BoHV-5. The aim of this study was to determine whether peripheral blood leukocytes and tonsils are targets for BoHV-5 infection and to establish whether all stages of that virus's infectious cycle can occur in those cell types. Comparisons with BoHV-1 infection of these tissues were also made in order to better understand the pathogenesis of both viruses. BoHV-1 and BoHV-5 were isolated from tonsils of acutely-infected calves. BoHV-5 was also isolated from a tonsil homogenate after dexamethasone-induced reactivation. During latency, infectious virus was recovered from a tonsil explant of one BoHV-5-infected calf. The genomes of BoHV-5 and BoHV-1 were detected in tonsils from acutely-infected calves although were not detected in tonsils from latently-infected calves or from calves treated with dexamethasone. Virus DNA was intermittently detected in leukocytes. The study has shown that BoHV-5 can establish latency in bovine tonsils and peripheral white blood cells, and that it can be reactivated from latently-infected tonsils, which might contribute to viral transmission. The titres of BoHV-1 and BoHV-5 in tonsils were similar, suggesting that replication at this site is a common feature for both viruses.

Asunto(s)

Infecciones por Herpesviridae/veterinaria , Herpesvirus Bovino 5/fisiología , Leucocitos/virología , Meningoencefalitis/veterinaria , Tonsila Palatina/virología , Latencia del Virus/fisiología , Animales , Bovinos , Enfermedades de los Bovinos/virología , Encefalitis Viral/veterinaria , Encefalitis Viral/virología , Infecciones por Herpesviridae/virología , Herpesvirus Bovino 1/fisiología , Herpesvirus Bovino 5/aislamiento & purificación , Ganglios Linfáticos/patología , Ganglios Linfáticos/virología , Meningoencefalitis/virología , Tonsila Palatina/patología

RESUMEN

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 (p = 0.05), and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05) 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.

Asunto(s)

Tonsila Faríngea/virología , Tonsila Palatina/virología , Tonsilitis/virología , Virosis/virología , Tonsila Faríngea/patología , Adenovirus Humanos/genética , Adolescente , Niño , Preescolar , Enfermedad Crónica , ADN Viral/genética , Femenino , Bocavirus Humano/genética , Humanos , Lactante , Masculino , Tonsila Palatina/patología , Virus ARN/genética , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Tonsilitis/epidemiología , Tonsilitis/genética , Tonsilitis/patología , Virosis/epidemiología , Virosis/genética , Virosis/patología

RESUMEN

This work studied the replication sites of hepatitis A virus (HAV) in cynomolgus monkeys (Macaca fascicularis) after intravenous inoculation. The cynomolgus monkeys were inoculated with the Brazilian hepatitis A virus strain (HAF-203). Monkeys were euthanized on days 15, 30, 45 and 60 postinoculation (pi). Liver samples, submandibular salivary gland, mesenteric lymph node and tonsils were removed for virological and pathological evaluation. Immunofluorescence analyses on liver and salivary gland sections using confocal laser scanning microscopy revealed the presence of HAV antigen (HAV Ag). The presence of HAV genome was monitored by real-time PCR. The HAV RNA was detected at 7 days postinoculation (dpi), concomitantly in serum, saliva and faeces. The highest HAV viral load was observed in faeces at 15 dpi (10(5) copies/ml), followed by serum viral load of 10(4) copies/ml at 20 dpi and saliva viral load of 10(3 )copies/ml at 7 dpi. The animals showed first histological and biochemical signs of hepatitis at 15 dpi. The HAV antigen (Ag) was present from day 7 until day 60 pi in the liver and salivary glands. The HAV replicative intermediate was also detected in the liver (4.5 x 10(4) copies/mg), salivary glands (1.9 x 10(3) copies/mg), tonsils (4.2 x 10(1) copies/mg) and lymph nodes (3.4 x 10(1) copies/mg). Our data demonstrated that the salivary gland as an extrahepatic site of early HAV replication could create a potential risk of saliva transmitted infection. In addition, the cynomolgus monkey was confirmed as a suitable model to study the pathogenesis of HAV human infection.

Asunto(s)

Virus de la Hepatitis A/patogenicidad , Hepatitis A/diagnóstico , Replicación Viral , Alanina Transaminasa/sangre , Animales , Modelos Animales de Enfermedad , Heces/virología , Técnica del Anticuerpo Fluorescente , Hepatitis A/patología , Hepatitis A/transmisión , Anticuerpos de Hepatitis A/sangre , Antígenos de Hepatitis A/aislamiento & purificación , Virus de la Hepatitis A/genética , Virus de la Hepatitis A/inmunología , Inyecciones Intravenosas , Hígado/enzimología , Hígado/virología , Ganglios Linfáticos/virología , Macaca fascicularis , Masculino , Microscopía Confocal , Tonsila Palatina/virología , ARN Viral/sangre , ARN Viral/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Saliva/virología , Glándulas Salivales/virología , Factores de Tiempo , Carga Viral

RESUMEN

To obtain information about the porcine circovirus type 2 (PCV2) infection status of pigs in Cuba and the probable association of PCV2 with other porcine viruses, tissue samples collected from ill pigs were evaluated using polymerase chain reaction (PCR). The PCR analysis showed that 67.7% of the samples (23/34) from seven swine herds of six different geographic regions were detected to be positive for PCV2. Ten of the 23 PCV2 positive samples (43.5%) shown a concurrent infection with porcine parvovirus (PPV) and 17 of 23 PCV2 positive samples (73.9%) exhibited a concomitant infection with classical swine fever virus (CSFV). This study is the first report of PCV2 infecting pigs with different clinical conditions in Cuban swine herds and provides evidence of PCV2 co-infection with PPV and CSFV in the field.

Asunto(s)

Infecciones por Circoviridae/veterinaria , Enfermedades de los Porcinos/virología , Animales , Infecciones por Circoviridae/diagnóstico , Infecciones por Circoviridae/patología , Circovirus/genética , Peste Porcina Clásica/patología , Peste Porcina Clásica/virología , Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Cuba , ADN Viral/genética , ADN Viral/aislamiento & purificación , Riñón/patología , Riñón/virología , Ganglios Linfáticos/patología , Ganglios Linfáticos/virología , Tonsila Palatina/patología , Tonsila Palatina/virología , Reacción en Cadena de la Polimerasa , Bazo/patología , Bazo/virología , Porcinos

RESUMEN

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.

Asunto(s)

Tonsila Faríngea/virología , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 8/aislamiento & purificación , Tonsila Palatina/virología , Tonsila Faríngea/química , Adolescente , Niño , Preescolar , ADN Viral/análisis , Femenino , Humanos , Hibridación in Situ , Lactante , Masculino , Tonsila Palatina/química , Adhesión en Parafina

RESUMEN

Human papilloma virus (HPV) is related to respiratory mucosal diseases, such as recurrent respiratory papillomatosis, as well as to upper-respiratory-tract malignancies. There are few reports concerning the prevalence of HPV in the upper respiratory tract of non-affected individuals. We examined the prevalence of HPV in the tonsils of children of the general population scheduled for tonsillectomy. Samples were taken from the tonsils of 100 children undergoing tonsillectomy and were then tested for HPV with the polymerase chain reaction (PCR) technique, utilizing the generic primers MY09 and MY 11. The study excluded children known to have HPV and HIV-related diseases. Parents and legal guardians completed a standardized socio-demographic questionnaire. The questionnaire revealed that 84 percent of the mothers had at least one risk factor for genital HPV. None of the tonsil samples were positive for HPV. Apparently HPV does not commonly colonize the tonsils of children undergoing routine tonsillectomy.

Asunto(s)

Adolescente , Niño , Preescolar , Femenino , Humanos , Masculino , Papillomaviridae/aislamiento & purificación , Tonsilectomía , Tonsila Palatina/virología , ADN Viral/análisis , Reacción en Cadena de la Polimerasa , Papillomaviridae/genética , Factores de Riesgo , Factores Socioeconómicos , Encuestas y Cuestionarios , Tonsila Palatina/cirugía

RESUMEN

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.

Asunto(s)

Tonsila Faríngea/virología , Herpesvirus Humano 8/aislamiento & purificación , Inmunohistoquímica/métodos , Hibridación in Situ/métodos , Tonsila Palatina/virología , Tonsila Faríngea/cirugía , Adolescente , Adulto , Niño , Preescolar , Femenino , Infecciones por Herpesviridae/virología , Humanos , Hipertrofia/cirugía , Lactante , Masculino , Tonsila Palatina/cirugía , Sarcoma de Kaposi/virología

RESUMEN

Human papilloma virus (HPV) is related to respiratory mucosal diseases, such as recurrent respiratory papillomatosis, as well as to upper-respiratory-tract malignancies. There are few reports concerning the prevalence of HPV in the upper respiratory tract of non-affected individuals. We examined the prevalence of HPV in the tonsils of children of the general population scheduled for tonsillectomy. Samples were taken from the tonsils of 100 children undergoing tonsillectomy and were then tested for HPV with the polymerase chain reaction (PCR) technique, utilizing the generic primers MY09 and MY 11. The study excluded children known to have HPV and HIV-related diseases. Parents and legal guardians completed a standardized socio-demographic questionnaire. The questionnaire revealed that 84% of the mothers had at least one risk factor for genital HPV. None of the tonsil samples were positive for HPV. Apparently HPV does not commonly colonize the tonsils of children undergoing routine tonsillectomy.

Asunto(s)

Tonsila Palatina/virología , Papillomaviridae/aislamiento & purificación , Tonsilectomía , Adolescente , Niño , Preescolar , ADN Viral/análisis , Femenino , Humanos , Masculino , Tonsila Palatina/cirugía , Papillomaviridae/genética , Reacción en Cadena de la Polimerasa , Factores de Riesgo , Factores Socioeconómicos , Encuestas y Cuestionarios

RESUMEN

BACKGROUND AND OBJECTIVES: Simian virus 40 (SV40) is an oncogenic DNA virus implicated in some human malignancies, including lymphomas. In the present masked case-control study, we investigated the prevalence of SV40 sequences and the expression of the viral oncoprotein, large tumor antigen (T-ag), in lymphomas and control specimens from patients negative for the human immunodeficiency virus in Costa Rica. DESIGN AND METHODS: Coded specimens were anlyzed by polymerase chain reaction for SV40 and Epstein-Barr virus (EBV). SV40 sequences were confirmed by Southern blot and DNA sequence analysis. Immunohistochemistry was used to detect the expression of SV40 T-ag in coded samples and to immunophenotype the lymphomas. RESULTS: When samples were decoded, SV40 DNA sequences were detected significantly more often in lymphomas than in control samples (30/125, 24% vs. 0/91, 0%; p=0.001). SV40 DNA was detected in 26% and 10% of non-Hodgkin's and Hodgkin's lymphomas, respectively. EBV DNA was detected in 10% of lymphomas and 33% of control specimens. None of the lymphomas was positive for both SV40 and EBV. Expression of SV40 T-ag was detected in 64% of B-cell lymphomas that contained T-ag DNA sequences and in none of the samples negative for viral DNA. Not all cells in a positive tumor expressed T-ag and the reactions were relatively low intensity. A germinal center B-cell-like profile was frequently associated with SV40-positive lymphomas. Of note, 20% of patients with SV40-related lymphomas were born in the 1970s and 1980s. INTERPRETATION AND CONCLUSIONS: These results indicate that SV40 is significantly associated with some B-cell neoplasms in Costa Rica today.

Asunto(s)

Antígenos Transformadores de Poliomavirus/análisis , Linfoma/virología , Vacuna Antipolio de Virus Inactivados/efectos adversos , Infecciones por Polyomavirus/epidemiología , Virus 40 de los Simios/patogenicidad , Infecciones Tumorales por Virus/epidemiología , Adulto , Estudios de Casos y Controles , Comorbilidad , Costa Rica/epidemiología , ADN Viral/análisis , Contaminación de Medicamentos , Infecciones por Virus de Epstein-Barr/epidemiología , Femenino , Centro Germinal/virología , Seronegatividad para VIH , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 4/patogenicidad , Enfermedad de Hodgkin/epidemiología , Enfermedad de Hodgkin/virología , Humanos , Neoplasias Hepáticas/epidemiología , Neoplasias Hepáticas/virología , Ganglios Linfáticos/virología , Linfoma/epidemiología , Linfoma de Células B/epidemiología , Linfoma de Células B/virología , Linfoma no Hodgkin/epidemiología , Linfoma no Hodgkin/virología , Masculino , Persona de Mediana Edad , Tonsila Palatina/virología , Infecciones por Polyomavirus/virología , Seudolinfoma/epidemiología , Seudolinfoma/virología , Virus 40 de los Simios/aislamiento & purificación , Neoplasias Gástricas/epidemiología , Neoplasias Gástricas/virología , Infecciones Tumorales por Virus/virología

RESUMEN

Recurrent tonsillitis has been the subject of much investigation. Events considered to predispose to or cause recurrent tonsillitis (RT) include the misuse of antibiotic therapy in acute bouts, alterations in the microflora, structural changes in crypt epithelium and certain viral infections. Epstein-Barr Virus (EBV) infection usually occurs in early childhood and can persist in palatine tonsil lymphocytes to induce tonsillitis at a later date. We have examined the presence of EBV in palatine tonsils in order to assess the relationship between this virus and recurrent acute tonsillitis. Tonsils were obtained from 85 patients, 2--14 years old (mean 5.6 years old) who underwent tonsils and adenoid (T&A) removal because of recurrent tonsillitis (RT) or T&A hypertrophy (TH). Tissues specimens were processed for non-isotopic in situ hybridization (ISH) using EBER 1/2 oligonucleotides (EBER RNA). The indications for surgery were RT in 42 patients and TH in 43 patients. In 25 out of 85 cases (29.4%) a positive EBER RNA reaction (15 RT and 33 TH) was found. The chi(2)-test showed no statistically significant difference in frequency of positive results between RT and TH group. We conclude that tonsils of children can be colonized by EBV and that the virus may be implicated in RT and TH.

Asunto(s)

Infecciones por Virus de Epstein-Barr/diagnóstico , Herpesvirus Humano 4/aislamiento & purificación , Tonsila Palatina/virología , Tonsilitis/patología , Tonsilitis/virología , Adolescente , Niño , Preescolar , Técnicas de Cultivo , Femenino , Humanos , Hibridación in Situ , Masculino , Reacción en Cadena de la Polimerasa , Recurrencia , Sensibilidad y Especificidad , Tonsilectomía , Tonsilitis/cirugía

RESUMEN

Burkitt's lymphoma (BL) and Hodgkin's disease (HD) occurring in developing regions are frequently associated with Epstein-Barr virus (EBV) infection and have a high incidence in childhood. Recent genotyping studies indicate that the tumour cells of both neoplasms represent B cells that contain somatically mutated immunoglobulin heavy chain genes. This implies that the precursors of these neoplasms have participated in the germinal centre (GC) reaction. We therefore presumed that normal lymphoid tissues from children living in developing regions would harbour an increased number of EBV-infected cells within the GC, when compared with children living in industrialized nations. To test this hypothesis, hyperplastic tonsils from 40 children living in Bahia (Brazil) and 40 from German children were analysed for the presence of EBV-encoded small nuclear RNA (EBER) and EBV-encoded proteins by in situ hybridization and immunohistology, respectively. Although the overall EBV infection rate was similar in both groups (50 per cent of Bahian vs. 45 per cent of German cases), a significantly higher number of EBER-positive lymphoid cells were found in the GCs of 8/20 EBV-positive tonsils from Brazil (9-89 cells/GC; mean: 14.5 cells/GC per case), while only 3/18 tonsils from Germany displayed a few EBER positive cells (1-9 cells/GC; mean: 0.5 cell/GC per case) in this compartment (p < 0.007). In addition, the EBV-infected GC cells in Bahian samples resembled centroblasts, exhibited mitotic activity, and in two cases showed expression of EBV-encoded latent membrane protein (LMP)-1, findings not present in German samples. These data show that latently EBV-infected cells participate more frequently in GC reactions in developing regions than in industrialized countries and may abnormally express the oncogenic protein LMP-1. This could in part explain the higher incidence in this region of EBV association with lymphomas related to GC cells or their progeny, such as BL and HD.

Asunto(s)

Infecciones por Virus de Epstein-Barr/patología , Centro Germinal/virología , Linfoma/virología , Tonsila Palatina/virología , Adolescente , Brasil , División Celular , Niño , Preescolar , Países en Desarrollo , Centro Germinal/patología , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 4/fisiología , Humanos , Técnicas para Inmunoenzimas , Hibridación in Situ , Tonsila Palatina/patología , ARN Viral/análisis , Latencia del Virus