Acometimento pulmonar e articular secundário à síndrome de Lemierre: relato de caso e revisão de literatura / Pulmonary and joint involvement secondary to Lemierre syndrome: case report and literature review

A síndrome de Lemierre caracteriza-se por uma rara entidade que gera tromboflebite da veia jugular interna e embolismo séptico em história da infecção recente da orofaringe, além de sinais radiológicos e isolamento de patógenos anaeróbicos, principalmente Fusobacterium necrophorum. Relatamos o caso de uma paciente do sexo feminino, 13 anos de idade, com histórico de carcinoma de nasofaringe associado ao vírus Epstein-Barr (estadiamento T4N2M0), submetida a procedimentos cirúrgicos e quimiorradioterapia. Iniciou com queixa de mialgia intensa, diplopia, lesões infectadas em membros e choque séptico. Por meio de exames de ultrassonografia cervical com Doppler colorido e tomografia computadorizada de pescoço com contraste endovenoso, foram identificados trombos intraluminais na veia jugular interna, além de trombos sépticos pulmonares, por meio da tomografia computadorizada de tórax. Posteriormente, ainda evoluiu com artrite piogênica coxofemoral esquerda. Foi isolada, por hemocultura, a bactéria Klebsiella pneumoniae Carpemenase, e o tratamento se deu pela associação entre vancomicina, amicacina, meropenem, metronidazol e anfotericina B. Conclui-se que, após o diagnóstico de SL e, embora com múltiplas complicações e diagnóstico tardio, a paciente encontra-se bem e assintomática, além do relato comprovar a dificuldade diagnóstica e de seu tratamento

Lemierre's syndrome is a rare condition that leads to thrombophlebitis of the internal jugular vein and septic embolism following recent oropharyngeal infection, being characterized by radiological signs and isolation of anaerobic pathogens, especially Fusobacterium necrophorum. We report the case of a 13-year-old female patient with history of nasopharyngeal carcinoma associated with Epstein-Barr virus (T4N2M0 staging), who underwent surgical procedures and chemoradiotherapy. Her initial complaint was severe myalgia, diplopia, infected limb injuries, and septic shock. Cervical color Doppler ultrasound and computed tomography scan of the neck with intravenous contrast showed intraluminal thrombi in the internal jugular vein, and chest computed tomography showed pulmonary septic thrombi. Subsequently, she progressed with left coxofemoral pyogenic arthritis. The bacterium Klebsiella pneumoniae Carpemenase was isolated in blood culture, and the patient was treated with the association of vancomycin, amikacin, meropenem, metronidazole, and amphotericin B. It is concluded that, despite the multiple complications and late diagnosis, the patient is well and asymptomatic after the diagnosis of Lemierre's syndrome; in addition, the report proves the difficulty of diagnosis and treatme

Leucemia agresiva de células natural killer en un paciente pediátrico / Agressive NK-cell leukemia in a pediatric patient

Resumen Introducción: Las neoplasias de células natural killer (NK) son poco frecuentes y representan <5% de todas las neoplasias linfoides. Comprometen diferentes entidades clínicas, como la leucemia de células NK, que es una neoplasia hematológica altamente agresiva con un pronóstico precario, que se presenta en hombres jóvenes y se observa con mayor frecuencia en ascendencia asiática. El virus de Epstein-Barr (VEB) parece estar relacionado con la patogenia de esta leucemia. Caso clínico: Se presenta el caso de un paciente de sexo masculino de 1 año y 7 meses de edad, quien inició su padecimiento con síndrome anémico, febril, infiltrativo e hiperleucocitosis. En el aspirado de médula ósea se detectaron blastos de morfología L2 (96%), inmunofenotipo CD56 (80.87%) y desoxinucleotidil transferasa terminal (84.11%). En la biopsia de médula ósea se identificó CD2+ membranoso, CD3+ citoplásmico y CD56+ membranoso; la serología para VEB fue positiva. El paciente recibió dos esquemas diferentes de quimioterapia basados en metotrexato, ifosfamida, etopósido, dexametasona y L-asparaginasa, y se documentó remisión parcial. Actualmente, se encuentra vivo con la enfermedad. Conclusiones: La leucemia de células NK es rara en adultos jóvenes, pero aún más en pacientes en edad pediátrica. Además, es de muy difícil tratamiento, ya que solo se cuenta con algunos reportes de casos, la sobrevida es de semanas a meses y las oportunidades de tratamiento se limitan. Recientemente, se ha evidenciado la utilidad del trasplante de médula ósea alogénico o células de cordón umbilical, y se ha logrado una sobrevida a 2 años. Las posibilidades terapéuticas en estos pacientes se encuentran en estudio. Se espera lograr en un futuro cercano la remisión completa y sobrevida a 5 años.

Abstract Background: Natural killer (NK) cell neoplasms are rare and represent <5% of all lymphoid neoplasms. They involve different clinical entities, of which one is NK cell leukemia, a highly aggressive hematologic neoplasm with poor prognosis that presents in young men and is more frequently seen in Asian descent. Epstein-Barr virus (EBV) seems to be related to the pathogenesis. Case report: A male patient of 1 year and 7 months of age, who began his condition with anemic, febrile, infiltrative syndrome and hyperleukocytosis is described. Bone marrow aspirate showed L2 morphology blasts (96%), CD56 (80.87%) and terminal deoxynucleotidyl transferase (84.11%) immunophenotype. Bone marrow biopsy showed membranous CD2+, cytoplasmic CD3+ and membranous CD56+; serology positive to EBV. The patient received two different chemotherapy schemes based on methotrexate, ifosfamide, etoposide, dexamethasone and L-asparaginase, which resulted in partial remission. Currently, the patient lives with the disease. Conclusions: NK cells leukemia is rare in young adults, but even more in pediatric patients, for which it is very difficult to treat because only a few cases have been reported in the literature, the survival varies from weeks to months and the chances of treatment are limited. Recently, the usefulness of allogeneic bone marrow transplantation or umbilical cord cells has been demonstrated, achieving a 2-year survival. The therapeutic possibilities in these patients are under study. In the near future, we hope to achieve the complete remission of the disease and a 5-year survival.

Análise comparativa do polimorfismo genético da glutationa transferase, do Helicobacter pylori e do vírus Epstein-Barr entre a área do tumor e as margens de ressecção proximal e distal do câncer gástrico / Comparative analysis of glutathione transferase genetic polymorphism, Helicobacter pylori and Epstein-Barr virus between the tumor area and the proximal and distal resection margins of gastric cancer

RESUMO Objetivo: comparar o polimorfismo dos genes Glutationa S-transferase teta 1 (GSTT1) e Glutationa S-transferase mu 1 (GSTM1) da área do tumor com as margens proximal e distal de espécimes de estômago ressecados de pacientes com câncer gástrico, e investigar a presença do DNA do vírus Epstein-Barr (EBV) e Helicobacter pylori. Métodos: coletamos prospectivamente amostras teciduais da área do tumor e das margens de ressecção proximal e distal dos estômagos de dez pacientes com adenocarcinoma gástrico submetidos à gastrectomia com linfadenectomia D2 e submetemos esses espécimes à extração de DNA. Comparamos a área do tumor com as margens proximal e distal dos estômagos ressecados para o polimorfismo dos genes GSTT1 e GSTM1 e investigamos a presença de DNA do EBV e H. pylori. Utilizamos o exon 5 do gene p53 como controle interno da reação de PCR multiplex. Resultados: em um paciente, detectamos genótipos GSTT1 e GSTM1 nulos na área do tumor, em contraste com a presença de ambos os genes nas margens proximal e distal. Encontramos DNA do EBV e H. pylori na área do tumor e também nas margens proximal e distal. Em outro paciente, a margem proximal foi negativa para GSTT1 e o DNA do EBV foi negativo na margem distal. Em três pacientes, o EBV-DNA foi negativo apenas na margem distal. Conclusão: este é o primeiro relato em que diferentes genótipos, infecção por EBV-DNA e H. pylori foram observados no mesmo paciente, indicando provável deleção desses genes em resposta à progressão tumoral e heterogeneidade intratumoral.

ABSTRACT Objective: to compare the polymorphism of the Glutathione S-transferase theta 1 (GSTT1) and Glutathione S-transferase mu 1 (GSTM1) genes from the tumor area with the proximal and distal margins of stomach specimens resected from patients with gastric cancer, and to investigate the presence of Epstein-Barr virus (EBV) DNA and Helicobacter pylori. Methods: we prospectively collected tissue specimens from the tumor area and from the proximal and distal resection margins of the stomachs of ten patients with gastric adenocarcinoma who underwent gastrectomy with D2 lymphadenectomy, and submitted these specimens to DNA extraction. We compared the tumor area with the proximal and distal margins of the resected stomachs for polymorphism of GSTT1 and GSTM1 genes and investigated the presence of EBV-DNA and H. pylori. We used the p53 exon 5 gene as an internal control of the multiplex PCR reaction. Results: in one patient, we detected null GSTT1 and GSTM1 genotypes in the tumor area, in contrast to the presence of both genes in the proximal and distal margins. We found EBV-DNA and H. pylori in the tumor area and also in the proximal and distal margins. In another patient, the proximal margin was negative for GSTT1, and EBV-DNA was negative in the distal margin. In three patients, EBV-DNA was negative only in the distal margin. Conclusion: this is the first report where different genotypes, EBV-DNA and H. pylori infection were observed in the same patient, indicating a probable deletion of these genes in response to tumor progression and intratumoral heterogeneity.

Epstein-Barr virus load in transplant patients: Early detection of post-transplant lymphoproliferative disorders / Carga de virus Epstein-Barr en pacientes trasplantados: detección temprana de desórdenes linfoproliferativos postrasplante

High levels of circulating EBV load are used as a marker of post-transplant lymphoproliferative disorders (PTLD). There is no consensus regarding the threshold level indicative of an increase in peripheral EBV DNA. The aim of the study was to clinically validate a developed EBV quantification assay for early PTLD detection. Transversal study: paired peripheral blood mononuclear cells (PBMC), plasma and oropharyngeal lymphoid tissue (OLT) from children undergoing a solid organ transplant with (n = 58) and without (n = 47) PTLD. Retrospective follow-up: 71 paired PBMC and plasma from recipients with (n = 6) and without (n = 6) PTLD history. EBV load was determined by real-time PCR. The diagnostic ability to detect all PTLD (categories 1-4), advanced PTLD (categories 2-4) or neoplastic PTLD (categories 3 and 4) was estimated by analyzing the test performance at different cut-off values or with a load variation greater than 0.5 log units. The higher diagnostic performance for identifying all, advanced or neoplastic PTLD, was achieved with cut-off values of 1.08; 1.60 and 2.47 log EBV gEq/10(5) PBMC or 2.30; 2.60; 4.47 log gEq/10(5) OLT cells, respectively. EBV DNA detection in plasma showed high specificity but low (all categories) or high (advanced/neoplastic categories) sensitivity for PTLD identification. Diagnostic performance was greater when: (1) a load variation in PBMC or plasma was identified; (2) combining the measure of EBV load in PBMC and plasma. The best diagnostic ability to identify early PTLD stages was achieved by monitoring EBV load in PBMC and plasma simultaneously; an algorithm was proposed.

La carga alta del virus Epstein-Barr se utiliza como un marcador de desórdenes linfoproliferativos postrasplante (post-transplant lymphoproliferative disorders [PTLD]). El objetivo de este estudio fue validar clínicamente un ensayo de cuantificación del virus Epstein-Barr para la detección temprana de PTLD. Se efectuó un estudio transversal en el que se analizaron muestras pareadas de células mononucleares periféricas (CMP), de plasma y de tejido linfoide orofaríngeo de niños con trasplante de órgano sólido, con PTLD (n = 58) y sin PTLD (n = 47). En el seguimiento retrospectivo se incluyeron 71 muestras pareadas de CMP y de plasma de trasplantados, con PTLD (n = 6) y sin PTLD (n = 6). La carga viral se determinó por PCR en tiempo real. Se estimó la capacidad diagnóstica para detectar PTLD (categorías: todas vs. avanzadas vs. neoplásicas) analizando diferentes valores de corte o una variación de carga mayor de 0,5 logaritmos. El mayor desempeño diagnóstico para identificar todos los PTLD, los avanzados y los neoplásicos, se obtuvo con valores de corte de 1,08; 1,60 y 2,47 log copias/10(5) en CMP y de 2,30; 2,60 y 4,48 log copias/10(5) en células de tejido linfoide orofaríngeo, respectivamente. La detección del ADN del virus Epstein-Barr en el plasma mostró una especificidad alta, pero una sensibilidad baja (todas las categorías) o alta (categorías avanzadas o neoplásicas) para identificar PTLD. Se observó el desempeño diagnóstico más alto en las siguientes condiciones: 1) al identificar una variación de carga en CMP o en plasma; 2) combinando la medición de la carga viral en CMP y en plasma. La mejor capacidad diagnóstica para identificar las etapas tempranas de los PTLD se logró mediante el seguimiento simultáneo de la carga viral en CMP y en plasma; se propone un algoritmo.

Establishment & characterization of lymphoblastoid cell lines from patients with multiple primary neoplasms in the upper aero-digestive tract & healthy individuals.

Background & objectives: A major drawback for genetic studies as well as long-term genotype-phenotype correlation studies in cancer is lack of representative human cell lines providing a continuous source of basic biomolecules and a system to carry out various experimental investigations. This can be overcome to some extent by establishing lymphoblastoid cell lines (LCLs) by infecting peripheral blood lymphocytes with Epstein Barr virus (EBV) which is known to immortalize human resting B cells in vitro giving rise to actively proliferating B-lymphoblastoid cell lines. The present study involves preparation and characterization of LCLs generated from patients with multiple primary neoplasms (MPN) of upper aero-digestive tract (UADT). Methods: Thirty seven LCLs were established from UADT MPN patients and healthy age, sex and habit matched controls using EBV crude stock. Characterization was done with respect to expression of CD-19 (Pan B-cell marker), CD3 (T cell specific marker), CD56 (NK-cell specific marker), cell morphology, ploidy analysis, genotype and gene expression comparison with the parent lymphocytes. Results: LCLs showed rosette morphology with doubling time of approximately 24 h. Ploidy analysis showed diploid DNA content which was maintained for at least 30 population doublings. When compared with parent lymphocytes there appeared no change at genetic and gene expression level. Interpretation & conclusions: Our results show that lymphoblastoid cell lines are a good surrogate of isolated lymphocytes bearing their close resemblance at genetic and phenotypic level to parent lymphocytes and are a valuable resource for understanding genotype-phenotype interactions.

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.

Frequent CD7 Antigen Loss in Aggressive Natural Killer-Cell Leukemia: A Useful Diagnostic Marker / 대한진단검사의학회지

BACKGROUND: Aggressive natural killer-cell leukemia (ANKL) is a rare neoplasm characterized by systemic proliferation of NK cells. However, the differential diagnosis of NK lymphoproliferative disorders is difficult because of the absence of a distinct diagnostic hallmark. Therefore, to identify diagnostic markers for ANKL, we analyzed the clinical data and laboratory findings obtained for 20 patients with ANKL. METHODS: From January 2000 to July 2007, 20 patients were diagnosed with ANKL on the basis of bone marrow studies. We retrospectively analyzed the clinical features and laboratory findings, including the complete blood count, Epstein-Barr virus status, immunophenotype, and the cytogenetic results. RESULTS: The subjects included 6 women and 14 men (median age, 44 yr; range, 2-70 yr). Cytogenetic studies were performed in 18 patients, and karyotypic abnormalities were observed in 9 patients (50%). None of the cytogenetic abnormalities were constantly observed in all the patients. However, 6q abnormalities were observed in 4 patients (4/18, 22%). The immunophenotype of the leukemic NK-cells was cytoplasmic CD3+, surface CD3-, CD16/56+, CD2+, and CD5-. Notably, the CD7 antigen was absent in 10 patients (50%). When the CD7 loss was combined with cytogenetic abnormalities, clonal markers could be identified in 75% of the ANKL cases. CONCLUSIONS: The CD7 antigen loss was frequently observed in our series of ANKL patients. In conjunction with the cytogenetic findings, this characteristic immunophenotypic finding can serve as a reliable marker for the timely diagnosis of ANKL. Therefore, immunophenotypic analysis of CD7 expression should be included in the diagnosis of NK cell neoplasms.

Epstein-Barr Virus-Positivity in Tumor has no Correlation with the Clinical Outcomes of Patients with Angioimmunoblastic T-cell Lymphoma

BACKGROUND/AIMS: Epstein-Barr virus (EBV) is involved in the pathogenesis of angioimmunoblastic T-cell lymphoma (AILT), but its precise role and prognostic impact are not clear. This study aimed to evaluate the incidence of EBV-postitivity in the tumor and bone marrow (BM) samples from AILT patients, and their correlations with the clinical variables and patient survival. METHODS: Seventy AILT cases were identified over a period of 8 years. Twenty seven cases were investigated for their EBV tumor status, and 10 BM samples of these patients were investigated for their EBV status with using in situ hybridization (ISH). EBV PCR was performed for the BM mononuclear cells in 8 cases. RESULTS: Among the 27 tumor specimens, ten (37%) were EBV-positive. Only CD20-negativity in tumor correlated with the EBV-positivity (p=0.035). In 13 (48%) patients, gross tumor involvement was recognized by hematoxylin-eosin staining at the time of diagnosis. Among the 10 patients who had additional BM slides available, there were 3 with BM involvement, and none showed EBV positive results on ISH. EBV PCR of the BM mononuclear cells revealed one-positive case among 8 patients. This patient was negative for both BM involvement and EBV ISH. The median overall survival of the 25 treated patients was 48.9 months (95% CI: 18.6~79.2 months). Neither overall survival nor progression-free survival was related with EBV-positivity of the tumor. CONCLUSIONS: EBV-positivity of tumor had no impact on the prognosis of AILT patients.

[Gastric adenocarcinoma with lymphoid stroma: about nine cases]

The purpose of this work is to study the pathological features and the outcome of lymphoid stroma gastric carcinoma and its relation with Epstein-Bar Virus [EBV] and the importance of the EBV serology in detection of early reccurence. Between January 1990 and december 2004, 155 patients underwent gastric resection for gastric carcinoma. Nine of them had lymphoid stroma gastric carcinoma. Over all survival as well as disease free survival were determined. Detection of reccurence was based on clinical exam and on findings yielded by endoscopy, radiological exams and EBV serology. A comparison of survival rate of patients beteween current gastric carcinoma and those with lymphoid stroma gatsric carcinoma was done using the Log Rank test. the patients [7 men and 2 women] had a mean age of 61 years [41 to 82]. The tumor was in the antrum in 4 cases, in the body of the stomach in 3 cases and in the cardia in 2 cases. The mean tumor size was 8 cm with deep invasion of the stomach wall in 5 cases and ganglion invasion in 6 cases out of 9. EBV was detected in all patients by in situ hybridation. Post operative morbidity and mortality were nil. Global survival and disease free survival were, respectively, 53.3% and 65.6% in lymphoid stroma gatsric carcinoma, and 43.4% and 63.5% for current gatsric carcinoma. Three patients died of a reccurent disease after 6, 30 and 33 months. The remaing 6 patients didn't have signs of reccurence. lymphoid stroma gastric carcinoma is a bulky, local disease. In our patients, it was always associated with EBV. Over all survival was better than survival of current gastric carcinoma, although there was no significance. EBV serology can be an effective screening procedure of reccurence

Carcinoma with Predominant Lymphoid Stroma in Hepatobiliary System: Report of 2 Cases / 대한간학회지

Lymphoepithelioma-like carcinoma (LELC) is an undifferentiated carcinoma with predominant lymphocytic infiltration, which is associated with Epstein-Barr virus (EBV) in variable proportions. We report two cases of carcinoma with predominant lymphoid stroma in hepatobiliary system. The first case was a lymphoepithelioma-like undifferentiated carcinoma with focal differentiation of cholangiocarcinoma (cytokeratin 19+) and hepatocellular carcinoma in light microscopy. The infiltration of CD8+ T lymphocytes was observed in the tumor and the surrounding hepatic parenchyme. In this tumor, EBV was detected and LMP1 was positive immunohistochemically. The second case showed the mixed features of lymphoepithelioma-like carcinoma and cholangiocarcinoma with predominant lymphoid stroma. In this case, EBV was detected. LELC of hepatobiliary system is an entity distinguished from conventional carcinoma with lymphoid stroma, and its association with EBV warrants further research.

Environmental factors related to gastric cancer associated with Epstein-Barr virus in Colombia.

Epstein-Barr virus (EBV)-encoded small RNA can be detected in about 1-17 % of gastric carcinomas. To elucidate lifestyles and other factors related to such an EBV-associated gastric carcinoma (EBV-GC), we conducted a case-control study in Cali, Colombia. The study subjects were 368 patients with gastric carcinoma newly diagnosed during the period between September 2000 and June 2003, including 42 EBV-GC cases. We obtained information on lifestyles, dietary habits, and occupational exposure by a questionnaire. The frequency of EBV-GC was related to birth order of patients (P for trend =0.025). More precisely, EBV-GC was much less frequent among the patients who were the eldest child in a family (P=0.007). Those findings were contrary to what was reported by the study conducted in Japan, where EBV-GC was more frequently observed among eldest brothers/sisters. A possible explanation for the apparently conflicting results is that EBV-GC risk is related to the age at first EBV infection but its relationship is not monotonic. In addition to the relationship with birth order, the present study showed that high salt intake and metal dust exposure may be related to EBV-GC as reported by the Japanese study although these associations observed in the present study were not statistically significant. No significant association was observed in other factors, including dietary habits. Further studies seem warranted to elucidate the difference between Japan and Colombia with respect to the environmental factors related to EBV-GC cases.

Epstein-Barr virus associated lymphoepithelial carcinoma of the parotid gland; a clinicopathological report of three cases.

Lymphoepithelial carcinoma is a relatively uncommon malignant tumor of the salivary gland demonstrating malignant epithelial cells with dense lymphoid stroma. The authors report three cases of lymphoepithelial carcinoma associated with Epstein-Barr virus of the right parotid gland with clinically presenting as painless, gradual enlargement of the preauricular mass. The histopathologic examination of the parotid gland is characterized by malignant epithelial cells with dense lymphoid stroma. Immunohistochemical stains show positive reactivity to cytokeratin and p53 in malignant epithelial cells. In situ hybridization of the Epstein-Barr virus-encoded Ribonucleic acid shows positivity in malignant epithelial cells. Clinical and pathologic features with relevant literatures are discussed. These are the first reported cases of primary parotid lymphoepithelial carcinoma associated with Epstein-Barr virus infection in Thailand and Southeast Asia.

Epstein-Barr virus-associated extranodal non-Hodgkin's lymphoma of the sinonasal tract and nasopharynx in Thailand.

Epstein-Barr virus (EBV) infection is highly associated with specific subtypes of malignant lymphoma. In our previous report on nodal malignant lymphoma in Thailand, we found that 64% of classical Hodgkin's lymphoma (cHL), 51% of non-Hodgkin's lymphoma, T-cell (NHL-T), and 13% of non-Hodgkin's lymphoma, B-cell (NHL-B) were EBV-related. In the present research, we conducted a retrospective study of primary extranodal non-Hodgkin's lymphoma of the sinonasal tract (e-NHL-ST) and primary extranodal non-Hodgkin's lymphoma of the nasopharynx (e-NHL-NP) in Southern Thailand, between 1997 and 2004. EBV-encoded RNA (EBER) expression by in situ hybridization was performed in all cases and a T-cell receptor (TCR)-g gene rearrangement study was performed in NHL-T cases. There were 18 cases of e-NHL-ST and 42 cases of e-NHL-NP detected by histologic and immunohistochemistry examinations. The percentages of e-NHL-ST and e-NHL-NP as compared to nodal malignant lymphoma were 3.7% and 6.8%, respectively. Sixteen cases (88.9%) of e-NHL-ST and 7 cases (16.7%) of e-NHL-NP were NHL-T, and the remainder were NHL-B. All of the NHL-T cases in both sites were EBER-positive. Two (5.4%) of the NHL-B cases in the nasopharynx showed EBER positive. Monoclonal bands of the TCR-gamma gene were detected in 71.4% of the extranodal NK/T-cell lymphomas, nasal type, patients; 50.0% of peripheral T-cell lymphoma, unspecified, patients; and one case of angioimmunoblastic T-cell lymphoma. This study indicates a very strong association of NHL-T in the sinonasal tract or nasopharynx with EBV infection, the link apparently being weaker in NHL-B patients. The study also indicates that most cases of extranodal NK/T-cell lymphoma, nasal type, are not the germline configuration of the TCR genes.

Prevalence of epstein barr virus in malignant lymphomas in Iraq

Malignant lymphomas are among the common tumors that are associated with and may complicate Epstein Barr Virus [EBV] infection. Although the strongest association is with the endemic Burkitt lymphomas [BL], the trend of association with sporadic lymhpomas reveals a consistently increasing prevalence of the virus in Hodgkin's disease [HD] in recent years compared to the non-Hodgkin type [NHL] which may point to a possible role for the virus in the predisposition and etio-pathogenesis of the disease. evaluate the association of EBV with Hodgkin's and non-Hodgkin lymphomas in relation to age, sex and HD subtype retrospectively using archival tissue biopsy sections. EBV was detected by In Situ Hybridization [ISH] using EBERs RNA probes in paraffin-embedded tissue sections prepared from archival tissue biopsy blocks. [a] EBV was detected in 25 of 40 HD cases [62.5%], 9 of 30 [30%] NHL cases, 4 of 10 [40%] BL cases, and in 5 of 20 [25%] other [non-BL] NHL cases, [b] Among the EBV-positive HD cases, 19 [76%] were of the mixed cellularity [MC] subtype, 1 [4%] of the Nodular Sclerosis [NS] subtype, 1 [4%] of the Lymphocyte Predominance [LP] subtype and 4 [16%] cases were of the Lymphocyte Depletion [LD] subtype, [c] Age distribution of HD cases revealed a bi-modal pattern characterized by an early major peak [67.5% of cases] below 35 years and a minor peak [32.5% of cases] above the age of 40. On the contrary, NHL cases revealed a nearly even age distribution [43.3% versus 56.6%] below and above the age of 40, respectively, [d] No difference was observed in the incidence of HD between males and females where the ratio was close to 1:1. On the other hand, a slight male predominance was seen among NHL cases with a male to female ratio of 2:1. [a] the prevalence rate of EBV infection was high among HD cases and fell within the prevalence rates found in previous similar studies revealing a range of values from 20 to 90%. [b] the higher prevalence of EBV positivity in HD compared to NHL found in this study points to a more substantial role for the virus in the pathogenesis of former compared to the latter disease which also comes in agreement with the greater environmental element compared to the genetic element in the etiology of HD. [c] The unexpected high EBV positivity in the LD subtype of HD may be interpreted as result of the progression of some of the early less aggressive MC-HD cases to the advanced more aggressive LD-HD subtype, [d] the bi-modal age distribution of EBV-positive HD cases follows the same pattern of distribution of the disease in general and testifies for the influence of environmental factors in the incidence of the disease

[Detection of EBV by PCR in fresh and paraffin-embedded naso pharyngeal carcinoma biopsies]

The nasopharyngeal carcinoma [NPC] is frequent in Tunisia. It's the second ORL cancer of men after the larynx one. To analyse the NPC characteristics in our population, we determined the frequency of EBV infection in 47 paraffin-embedded and 6 fresh NPC biopsies. We first extracted the DNA from tumoral tissus and then amplified viral sequences by PCR to detect and to type the infecting virus [EBV-A or ABV-B]. Our results showed that amplifiable DNA has been obtained from 34/47 paraffin-embedded NPC biopsies while 13/47 of the others biopsies contained degraded and not amplifiable DNA. All the fresh biopsies allowed to obtain DNA with good quality. The EBV infection frequency in paraffin-embedded NPC biopsies is 35% while EBV is detected in all fresh biopsies [6/6]. Our analyse also showed that the EBV-A is predominant in our population compared to EBV-B as it was shown in most countries of the world. This study clearly shows that PCR results obtained with paraffin-embedded NPC biopsies are divergeant from those obtained with fresh biopsies. Because of DNA degradation in paraffin-embedded NPC biopsies, the biology molecular results from that kind of samples is criticable. Moreover, the results obtained from fresh NPC biopsies confirmed the quasi-constant association of EBV with undifferenciated carcinoma nasopharyngeal type

Extranodal malignant lymphoma of the upper aerodigestive tract: prevalence of Epstien-Barr virus (EBV) infection in King Chulalongkorn Memorial Hospital.

BACKGROUND: High frequency of Epstein-Barr virus (EBV) in the normal mucosa of the upper aerodigestive tract suggests that it may serve as a reservoir for the virus. Malignant lymphomas arising in this site may be associated with EBV. OBJECTIVES: To determine the prevalence of EBV infection in extranodal malignant lymphomas of the upper aerodigestive tract. SETTING: King Chulalongkorn Memorial Hospital, Thailand. DESIGN: Descriptive study. PATIENTS: 42 Thai patients who presented between 1998 and 2003. MATERIAL AND METHOD: The expression of EBV mRNAs (EBERs) of malignant lymphoma was studied by means of in situ hybridization in formalin-fixed, paraffin-embedded specimens. RESULTS: The recruited subjects were 26 males and 16 females, and their age ranged from 3 to 85 years with the mean of 51.43 years, in 4 of them human immune deficiency virus (HIV) infection was documented. Ten of 42 cases (23.81%) expressed EBER transcripts and were extranodal NK/T-cell lymphomas, nasal type (7 cases), plasmablastic lymphomas (2 cases) and diffuse large B-cell lymphoma (1 case). Three of 4 cases (75%) of known HIV-seropositive cases were EBV-positive (2 plasmablastic lymphomas and 1 diffuse large B-cell lymphoma). CONCLUSION: In the upper aerodigestive tract, EBV was present in some but not all malignant lymphoma. It was associated with extranodal NK/T-cell lymphoma, nasal type and B-cell lymphoma arising in HIV-infected patients, but it was not found in B-cell lymphoma arising in immunocompetent patients.

Establishment of real-time polymerase chain reaction-based assay for quantitation of Epstein-Barr virus DNA in healthy donors and in patients with EBV associated lymphoma.

Epstein-Barr virus (EBV) is associated with several malignancies including nasopharyngeal carcinoma and lymphoma in immunocompromised patients. Quantitative monitoring of EBV DNA in these patients has recently become essential for management of the disease. In the present study the authors developed a rapid and reliable real-time PCR to quantify the EBV DNA in peripheral blood mononuclear cell (PBMC) using hybridization probe technique. The real-time primers and probes in this real-time PCR system were designed based on EBNA-1 sequence. The newly-established real-time PCR demonstrated its high sensitivity (as few as 10 copies of EBV could be detected) and specificity. The intra- and inter-assay variations of the assay were shown to be within a 0.5-log10-difference range. A total of 2 EBV-seronegative, 14 EBV-seropositive healthy donors and 4 patients with PCNSL were enrolled into the study. Our results revealed the median of EBV-DNA in lymphoma patients (7886 copies/10(6) PBMC or 15,150 copies /microg DNA) was higher than that of healthy donors (<10 copies/l0(6) PBMC or <10 copies/microg DNA) with statistic significance (P < 0.01). Assessment of this assay in larger number of donors and patients will provide clinical cut-off values which are essential for monitoring and diagnosis of EBV-associated diseases.

Association of the Epstein-Barr viruses with Hodgkin lymphoma: an analysis of pediatric cases in Thailand.

An investigation as to whether any association of pediatric HL in Thailand was likely to be EBV positive was performed on formalin-fixed paraffin embedded tissue sections using in situ hybridization for EBV encoded RNA (EBER) technique. The analysis was performed on 15 cases. They were 11 male and 4 female cases. The subtypes of HL according to WHO classification were nodular lymphocyte predominance in 1 (6.6%), nodular sclerosis in 4 (26.6%), mixed cellularity in 9 (60%) and lymphocyte depletion in 1 (6.6%). EBV encoded RNA by in situ hybridization was demonstrated in 92.8% of classic HL: 3 of 4 (75%) with nodular sclerosis; 9 of the 9 with mixed cellularity (100%) and 1 of 1 (100%) with lymphocyte depletion. Case of nodular lymphocyte predominance was negative for EBV, CD 15 CD 30 and positive for CD 20. CD 15 and CD 30 were positive in 78.6% and 85% respectively for classic HL. Our results suggest a strong association of EBV with pediatric classic HL (92.3%) particularly the mixed cellularity subtype (100%). The result confirms the male predominance in pediatric HL. Mixed cellularity is the most common subtype of HL in our series (60%).

Nasal NK/T cell lymphoma mimicking a squamous cell carcinoma: a case report.

A 48 year old female presented with extensive ulceration of the nasal septum of 8 months duration. Investigations confirmed the local nature of the disease. A biopsy revealed large zones of ischemic necrosis and abnormal lymphoid cells invading vessel walls and glandular structures. Florid squamous metaplasia, and pseudoepitheliomatous hyperplasia of mucosal epithelium mimicked squamous cell carcinoma and necrotising sialometaplasia. Immunohistochemistry and insitu hybridization confirmed the diagnosis of an EBV positive, Nasal NK/T cell lymphoma. A Pubmed/Medline search suggests that this is the first documented case from India.

Viral infections of renal allografts--an immunohistochemical and ultrastructural study.

Viral infections have been recognized as an integral part of both graft injury and rejection. On routine histology, viral infections are diagnosed only when fully established, by the presence of viral inclusions or cytopathic effect. Although renal transplants are routinely done in many centres in India, the incidence of viral infections is largely unkown. This study was aimed at detecting 5 viral infections namely, cytomegalovirus (CMV), BK polyoma Virus (BKV), Herpes Simplex Virus1 and 2 (HSV1 and 2) and Epstein Barr Virus (EBV) in renal biopsies from 321 renal allograft patients, using immunohistochemical and electron microscopic studies. Sixty two biopsies were selected from a total of 414 (belonging to 321 patients) for immunostaining on the basis of features suspicious of viral infections in hematoxylin and eosin stained sections. Immunostaining confirmed CMV infection in 8 biopsies, BKV infection in 31 biopsies and HSV1 in only 2 biopsies. HSV2 and EBV were not detected in any biopsy. Two biopsies showing CMV immunopositivity and 5 of BKV were further processed for electron microscopy, which supported the diagnoses. Thus, the study highlights the prevalence of BKV and CMV infections in renal transplant patients having graft dysfunction, to be 9.3% and 1.9%, respectively. Besides, only one case each was diagnosed as CMV infection and BKV infection in routine histopathological reporting, establishing the importance of immunohistochemical studies in early diagnosis of these viral infections.