Human papillomavirus and Epstein Barr virus in oral hairy leukoplakia among HIV positive Venezuelan patients

Oral hairy leukoplakia (OHL) is commonly found in individuals infected with HIV, and represents the most frequent oral manifestation. The purpose of this study was to detect the presence of Human Papillomavirus (HPV) and Epstein Barr Virus (EBV) in OHL of HIV+ Venezuelan patients. We evaluated 21 HIV+ adult patients with clinically present OHL lesions: 11 under antiretroviral therapy, 10 without therapy, and 10 oral mucosal samples as controls. Nested-PCR was used to detect EBV and HPV infection. The INNO-LiPA HPV Genotyping v2 was applied to determine the HPV genotype. The EBV genome was found in 16/21 (76%) of the HIV+ patients with OHL. No difference was observed in EBV+ and EBV- patients related to antiretroviral therapy viral load and CD4+ T cell count. HPV-DNA was observed in 7/21 HIV positive cases (33%). The HPV genotypes detected were: 6, 11, 31, 33, 52, and 56/74. The most frequently HPV found was genotype 6 in 7/7, while two cases were HPV-11 and two HPV-52. Of the positive cases, 5/7 (71%) presented co-infection with more than one HPV genotype and 4/7 (57%) had HPV coinfection with high and low risk types. No case was EBV or HPV positive in the control group. In this study, a higher EBV prevalence was observed in OHL-HIV+ patients, confirming the etiologic role in this entity. A considerable number of cases were positive for HPV infection, and many patients presented coinfection with more than one HPV genotype as well as the presence of high oncogenic risk HPV in OHL.

El proposito del presente estudio fue detectar la presencia de virus papiloma humano (VPH) y Epstein Barr (VEB) en Leucoplasia Vellosa Oral (LVO) de pacientes VIH positivos. Se evaluaron 21 pacientes adultos VIH positivos con lesiones clinicas presentes de LVO y 10 casos controles de mucosa sana. Para el diagnostico molecular de VPH y EBV se utilizo Nested PCR. La determinacion de los genotipos se realizo mediante el kit HPV INNO-LiPA genotyping v2. La presencia de genoma de VEB se demostro en un alto porcentaje (76%) en 16/21 de los pacientes VIH positivos con LVO. No se observo relacion entre los pacientes VEB+ y VEBcon el uso de terapia antirretroviral, la carga viral y el contaje de celulas T CD4+. Se demostro la presencia de ADN-VPH en 7/21 (8%) de los casos VIH positivos. Los genotipos de VPH detectados fueron 6, 11, 31, 33, 52, 56/74. El genotipo 6 fue el mas frecuentemente observado en 7/7, dos casos fueron VPH 11 y dos VPH 52. De los casos positivos 5/7 (71%) presentaron coinfeccion con mas de un genotipo de VPH y en 4/7 (57%) se evidencio coinfeccion con tipos de alto y bajo riesgo oncogenico. En el presente estudio se observo una alta prevalencia de VEB en pacientes VIH positivos con LVO, confirmando el papel etiologico en esta entidad. Un considerable numero de casos fueron positivos para VPH. Se observo la presencia de coinfeccion con mas de un tipo viral, asi como la presencia de VPH de alto riesgo.

Polymerase chain reaction genotyping of Epstein-Barr virus in scraping samples of the tongue lateral border in HIV-1 seropositive patients

The Epstein-Barr virus (EBV) is the etiological agent of oral hairy leukoplakia (OHL), an oral lesion with important diagnostic and prognostic value in acquired immunodeficiency disease syndrome. The two EBV genotypes, EBV-1 and EBV-2, can be distinguished by divergent gene sequences encoding the EBNA-2, 3A, 3B, and 3C proteins. The purpose of this study was to identify the EBV genotype prevalent in 53 samples of scrapings from the lateral border of the tongue of HIV-1 seropositive patients, with and without OHL, and to correlate the genotypes with presence of clinical or subclinical OHL with the clinic data collected. EBV-1 and EBV-2 were identified through PCR and Nested-PCR based on sequence differences of the EBNA-2 gene. EBV-1 was identified in the 31 samples (15 without OHL, 7 with clinical OHL and 9 with subclinical OHL), EBV-2 in 12 samples (10 without OHL, 1 with clinical and 1 subclinical OHL), and a mixed infection in 10 samples (2 without OHL, 3 with clinical and 5 with subclinical OHL). The presence of EBV-1 was higher in women, but a significant statistical result relating one the EBV genotypes to the development of OHL was not found. We conclude that the oral epithelium in HIV-1 seropositive patients can be infected by EBV-1, EBV-2 or by a mixed viral population.

Prevalence of oral hairy leukoplakia and epithelial infection by Epstein-Barr virus in pregnant women and diabetes mellitus patients: cytopathologic and molecular study

Oral hairy leukoplakia (OHL) is generally reported in patients with severe immunosuppression, except for a few cases in individuals with moderate degree of immunodeficiency. It is a white lesion that appears mainly in the lateral border of the tongue, caused by Epstein-Barr virus (EBV). The nuclear changes caused by EBV (Cowdry A inclusion, ground glass and nuclear beading), observed in cytopathology, are specific and enough for the definitive diagnosis of OHL, independent of the identification of the virus. Here we investigated the prevalence of OHL and the presence of EBV-DNA in the lateral borders of the tongue from 90 pregnant women, 90 diabetes mellitus (DM) patients, 30 healthy individuals (negative group) and 30 HIV+ with OHL (positive group). Smears were analyzed by cytopathology and polymerase chain reaction (PCR). A case of subclinical OHL and candidiasis was identificated in a DM patient by cytopathologic analysis. PCR results demonstrated EBV-DNA in 65 percent of the pregnant women, in 35 percent of DM patients, and in 20 percent of the healthy individuals. We concluded that DM patients can develop OHL with a low prevalence. Furthermore, the prevalence of the EBV in lateral border of the tongue is larger in pregnant women than in healthy individuals.

Prevalence of oral hairy leukoplakia in 120 pediatric patients infected with HIV-1

A leucoplasia pilosa oral (OHL) é uma infecção oportunista causada pelo Vírus Epstein-Barr (EBV) encontrada em pacientes infectados pelo HIV. É uma lesão assintomática que tem um importante valor prognóstico na AIDS. Diferentemente de pacientes adultos, a OHL tem sido descrita na literatura como tendo uma prevalência muito pequena em pacientes pediátricos. Logo, o objetivo deste estudo foi investigar a prevalência da OHL em pacientes pediátricos positivos para o HIV através do uso da citopatologia. A amostra consistiu-se de 120 pacientes, que foram submetidos a exame oral e coleta de material de ambos os lados da língua. O critério diagnóstico foi baseado na identificação de alterações nucleares. A OHL clínica foi identificada em dois (1,67%) pacientes. A citopatologia revelou vinte casos (16,7%) de OHL subclínica. Nossos resultados mostram que a prevalência de OHL em pacientes pediátricos infectados pelo HIV deve ser maior que a relatada na literatura.

Epstein-Barr virus in oral hairy leukoplakia scrapes: identification by PCR

A leucoplasia pilosa (LP) é uma lesão associada ao comprometimento do sistema imune e seu diagnóstico é determinado pela demonstração da presença do vírus Epstein-Barr (EBV) no tecido lesionado. O objetivo deste trabalho foi desenvolver uma metodologia simples para auxiliar no diagnóstico da LP, utilizando-se a técnica da PCR como uma alternativa para evidenciar o EBV em esfregaços da lesão. Amostras de DNA foram obtidas por meio de raspado de borda lateral de língua, de 38 pacientes adultos, sendo estes: 29 pacientes HIV positivos (4 com evidência clínica de leucoplasia pilosa, 4 que haviam apresentado LP previamente, mas não no momento da coleta, e 21 sem evidência clínica de leucoplasia pilosa) e 9 voluntários sadios para o grupo controle. O DNA foi extraído do material obtido por raspagem e amplificado pela PCR utilizando-se iniciadores específicos para o EBV. Dos 29 casos de pacientes portadores do vírus HIV, 22 (75,86%) foram positivos para o EBV, sendo: 2 de pacientes com evidência clínica de LP, 4 de pacientes que haviam apresentado LP previamente e 16 de pacientes sem sinais clínicos de LP. No grupo controle, as amostras de 5 (55,56%) indivíduos clinicamente sadios mostraram amplificação para o EBV. Concluímos que o uso da PCR em esfregaços sugere uma alta sensibilidade e baixa especificidade no diagnóstico da leucoplasia pilosa.