Investigação da presença e da influência do Epstein-Barr vírus na severidade da papilomatose laríngea / Investigation of the presence and influence of Epstein-Barr virus on the severity of laryngeal papilomatosis

A papilomatose laríngea é uma neoplasia benigna causada pelo papilomavírus humano (HPV), sendo os tipos 6 e 11 os mais comuns, e que ocorre em dois grupos etários, juvenil e adulto. A possível coinfecção viral tem sido sugerida em lesões de cabeça e pescoço; nesse sentido, o Epstein Barr vírus (EBV), que também apresenta tropismo por células epiteliais vem sendo estudado neste grupo de lesões. Os objetivos deste estudo foram genotipar os HPVs, investigar a presença de EBV-DNA por PCR e EBV-RNA por hibridização in situ. Além disso, associar a presença de EBV com a imunoexpressão de CD21, os resultados obtidos com a escala laringoscópica de Derkay et al. (1998) e com os dados clinicopatológicos. Oitenta casos de papilomatose laríngea, juvenil (n=36) e adulta (n=44), foram retrospectivamente analisados e subdivididos em grupos de menor e maior severidade, baseando-se na escala de Derkay. Todas as amostras foram HPV posivitas, com 49 casos HPV 6, 26 casos HPV 11, 4 casos HPV 6 e 11, e 1 caso HPV 16. A presença de EBV-DNA foi detectada em 9 amostras, entretanto EBV-RNA não foi não foi identificado em nenhuma amostra. Assim como a presença do EBV-DNA, a imunoexpressão de CD21 não se associou estatisticamente com quaisquer variáveis. A presença de HPV 6 foi mais comum em PLA e, o HPV 11 foi mais comum (p=0,02) e maior em casos de maior severidade (p=0,04), no grupo juvenil. A presença do EBV provavelmente não desempenha papel importante na progressão/severidade desta patologia(AU)

Laryngeal papillomatosis is a benign neoplasm caused by the human papillomavirus (HPV), been types 6 and 11 the most commonly related, and is divided into two groups: juvenile and adult. Viral coinfection has been suggested in head and neck lesions; in this sense, Epstein Barr virus (EBV), which also presents tropism for epithelial cells, has been studied in this group of lesions. The aims of this study are to perform HPV genotyping, investigate EBVDNA presence by PCR and EBV-RNA by in situ hybridization; and associate EBV presence with CD21 immunoexpression. Finally, the results were associated with Derkay laryngoscopic score. Eighty cases of laryngeal papillomatosis, juvenile (n = 36) and adult (n = 44) were retrospectively subdivided into low-risk and high-risk of severity based on the Derkay index. All samples were HPV-positive, with 49 cases of HPV 6, 26 cases of HPV 11, 4 cases of HPV 6 and 11, and 1 case of HPV 16. The presence of EBV-DNA was detected in 9 samples, however EBV-RNA was not identified in any sample. As the presence of EBV-DNA, the CD21 immunoexpression was not statistically associated with any variables. The presence of HPV 6 was more common in ALP, HPV 11 was more common (p = 0.02) and higher in cases of higher severity (p = 0.04) in juvenile group. The presence of EBV probably does not play an important role in the progression/severity of this pathology(AU)

Infección por virus de Epstein-Barr y hepatitis aguda colestásica / Infection with Epstein-Barr virus and acute cholestatic hepatitis

El Virus de Epstein-Barr (VEB) es un herpes virus cuyo medio de transmisión es a través de secreciones de una persona portadora del virus, siendo el hombre el único huésped. La primo infección por lo general es asintomática o puede manifestarse como mononucleosis infecciosa con la triada clásica de fiebre, faringitis y adenopatías. Esta cursa con elevación leve y autolimitada de transaminasas, por lo que solo un 5% de los casos se ha asociado con hepatitis aguda colestásica. Presentamos a un paciente con una infección por virus de Epstein-Barr y hepatitis aguda colestásica con historia de aparición de una masa cervical lateral derecha. Al examen físico evidencia ictericia a nivel de escleras, mucosas y ambos miembros superiores. Niveles de bilirrubina en sangre elevados. Paciente con ultrasonido hepático y vías biliares normal, colangiopancreatografía retrograda endoscópica normal por lo que se procede a realizar pruebas serológicas para VEB siendo esta positiva. Se da tratamiento con ganciclovir, mejorando pruebas de función hepática y disminuyendo ictericia, teniendo así una evolución favorable del paciente...(AU)

Epstein-Barr Virus (EBV) is a herpes virus, whose means of transmission is through secretions of a person carrying the virus, the man being the only host. The cousin infection is usually asymptomatic or may manifest as infectious mononucleosis with the classical triad of fever, pharyngitis and lymphadenopathy. This is a mild and self-limiting elevation of transaminases, which means that only 5% of the cases have been associated with acute cholestasis hepatitis. We present a patient with an Epstein-Barr virus infection and acute cholestasis hepatitis with a history of the appearance of a right lateral cervical mass. Physical examination shows jaundice at the level of sclera, mucosa and both upper limbs. Elevated blood bilirubin levels. Patient with hepatic ultrasound and normal bile ducts, normal endoscopic retrograde cholangiopancreatography, so serological tests for EBV are performed and this is positive. Ganciclovir is given, improving liver function tests and decreasing jaundice, thus having a favorable evolution of the patient...(AU)

Detection and Typing of HSV-1, HSV-2, CMV and EBV by Quadruplex PCR

The development of a multiplex polymerase chain reaction (PCR) method for rapid and accurate detection and typing of herpes simplex virus type 1 (HSV-1), and type-2 (HSV-2), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) is very important for clinical diagnosis to allow the deliver of therapy as early as possible. Large scale amplifications by multiplex PCR of viral DNA can lower the cost and time for viral diagnosis. In this study, therefore sensitive quadruplex PCR was achieved by optimizing parameters such as primers, and 1.5 mM magnesium and 200 uM dNTPs concentrations. The concentrations of HSV-1, HSV-2, CMV and EBV primers were 0.5, 0.3, 0.25 and 0.25 pmoles, respectively. Optimal annealing temperature was 54 degrees C. Employing these conditions, we could detect 10 copies of reconstructed template plasmid DNA, which were cloned to vectors containing target sequences of viral DNA. PCR products of 271 bp for HSV-1, 231 bp for HSV-2, 368 bp for CMV, and 326 bp for EBV were separated on 5.0% polyacrylamide gel electrophoresis and confirmed by direct sequencing. The present study showed that the quadruplex PCR assay described herein has potential application in clinical diagnosis, when rapid, accurate detection and typing of viruses HSV-1, HSV-2, CMV or EBV are necessary.

Epstein-Barr virus in nasopharyngeal carcinoma in Indian patients.

Formalin fixed paraffin embedded tissue blocks from 40 cases of nasopharyngeal carcinoma were investigated for the presence of Epstein-Barr virus (EBV). Heminested polymerase chain reaction was employed to amplify U 2 region of EBV DNA coding for EBNA 2 gene. This is the first study of its kind carried out in India. EBV was detected in 28 out of 40 (70%) nasopharyngeal carcinomas. Type A EBV was identified in 20 cases (71.4%), type B in 7 cases (25%) and coinfection in one case (3.5%). Either type A, or type B EBV, was identified in 22 out of 23 cases (95.7%) of undifferentiated nasopharyngeal carcinoma and 6 out of 15 cases (40%) of non keratinized squamous carcinoma. The higher incidence of EBV in non keratinized squamous carcinoma of nasopharynx assumes importance in view of the contrary reports in other studies. 26 cases were from Western India, of which 15 (57.6%) showed presence of type A EBV and 2 (7.6%) type B EBV. From Eastern Indian particularly Assam, 4 our of 5 cases showed positivity for type B EBV and 1 a coinfection. Whereas, from Bihar 3 out of 4 cases showed presence of type A EBV. From Middle East (Arabs) type A EBV was detected in 3 out of 5 cases of nasopharyngeal carcinomas. Geographically, type A EBV was far more prevalent in Western India, while in Eastern India particularly Assam, all five vases were positive for type B EBV. Thus, a significant variation in the type of EBV infection was observed in nasopharyngeal carcinoma in different ethnic populations in India.

Detection of Epstein-Barr virus DNA and HHV-6 DNA in tissue biopsies from patients with nasopharyngeal carcinoma by polymerase chain reaction.

A total of 34 tissue biopsies were collected from nasopharyngeal carcinoma (NPC) patients and 5 controls with non-NPC. Extracted DNA from tissue biopsies were analyzed for presence of specific gene sequences to EBV type A and type B, and HHV-6 by polymerase chain reaction (PCR). The different sequences of EBV type A and B were parts from the highly divergent forms of the EBV nuclear antigen 2 (EBNA 2). The PCR amplified products for EBNA 2A and EBNA 2B were 115 and 119 base pairs respectively whereas that of HHV-6 DNA was 776 base pairs. The results demonstrated that EBV DNA was detected in 32 of 34 cases (94.1%): 28 (82.3%) with type A, 2 (5.9%) with type B, and 2 (5.9%) with both types. EBV DNA of type A could be detected 1 (20%) of 5 controls. HHV-6 DNA was in 5 of 34 samples (14.7%) whereas HHV-6 DNA was not detectable in biopsy tissues from controls. The results show that in the NPC patient group, A type of EBV is predominant. Detection of HHV-6 DNA in patients group only might be resulted from reactivation of a latent infection or association with EBV-induction of NPC.