RESUMEN
OBJECTIVES: To develop and validate a prognostic nomogram based on baseline nutritional and inflammatory parameters for risk stratification in patients with de novo metastatic nasopharyngeal carcinoma (dmNPC) receiving chemotherapy combination programmed death-1 (PD-1) inhibitor. METHODS: This retrospective study analyzed 131 patients with dmNPC (88 and 43 in the training and validation cohorts, respectively) between March 2017 and November 2020. All these patients received chemotherapy combined with PD-1 inhibitor treatment. We identified independent risk factors using univariate and multivariate Cox regression analyses and established a nomogram to predict the progression-free survival (PFS). The predictive accuracy of the nomogram was evaluated and independently validated. RESULTS: Baseline nutritional risk index (NRI), prognostic nutritional index (PNI), systemic immune-inflammation index (SII), uric acid (UA), and post-treatment Epstein-Barr virus (EBV) DNA were used to develop a nomogram that could divide patients into favorable- and unfavorable-prognosis groups. The median PFS (mPFS) was significantly longer in the favorable-prognosis group compared to the unfavorable-prognosis group (35.10 months [95% CI: 27.36-42.84] vs. 7.23 months [95% CI: 6.50-7.97]; p = 0.001). All results were confirmed in the validation cohort. CONCLUSIONS: The proposed model improved the prognostic risk stratification for patients with dmNPC undergoing chemotherapy combined with PD-1 inhibitor treatment.
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Infecciones por Virus de Epstein-Barr , Neoplasias Nasofaríngeas , Humanos , Carcinoma Nasofaríngeo/tratamiento farmacológico , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/patología , Herpesvirus Humano 4/genética , Inhibidores de Puntos de Control Inmunológico/efectos adversos , Estudios Retrospectivos , Infecciones por Virus de Epstein-Barr/complicaciones , Neoplasias Nasofaríngeas/tratamiento farmacológico , Neoplasias Nasofaríngeas/patología , Pronóstico , ADN ViralRESUMEN
Space exploration has brought many challenges to human physiology. In order to evaluate and reduce possible pathological reactions triggered by space environments, we conducted bioinformatics analyses on the methylation data of the Mars 520 mission and human transcriptome data in the experiment simulating gravity changes. The results suggest that gene expression levels and DNA methylation levels were changed under the conditions of isolation and gravity changes, and multiple viral infection-related pathways were found in the enrichment analysis results of changed genes including Epstein Barr virus (EBV) infection, Hepatitis B virus (HBV) infection, Herpes simplex virus (HSV) infection and Kaposi's sarcoma-associated herpesvirus (KHSV) infection. In this study, we found that Epigallocatechin-3-gallate (EGCG) and vitamin D are helpful in reducing viral infection risk. In addition, the causal associations between nutrients and viral infections were calculated using Two sample Mendelian Randomization (2SMR) method, the results indicated that vitamin D can reduce EBV infection and HBV infection risk. In summary, our study suggests that space environments increase the risk of human viral infection, which may be reduced by supplementing EGCG and vitamin D. These results can be used to formulate medical plans for astronauts, which have practical application value for future space exploration.
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Infecciones por Virus de Epstein-Barr , Suplementos Dietéticos , Infecciones por Virus de Epstein-Barr/complicaciones , Infecciones por Virus de Epstein-Barr/genética , Herpesvirus Humano 4/genética , Humanos , Nutrientes , Vitamina D/genética , Vitamina D/uso terapéuticoRESUMEN
BACKGROUND AND AIMS: Limited data are available for tumor immune microenvironment (TIME) in Epstein-Barr virus (EBV)-associated lymphoepithelioma-like cholangiocarcinoma (EBV-LELCC), a rare subtype of intrahepatic cholangiocarcinoma (IHCC). We aimed to investigate TIME features in EBV-LELCC and the correlation between the components of TIME and the clinical outcomes. METHODS: Tumor tissues from five EBV-LELCC cases confirmed through EBER in situ hybridization and five stage-matched conventional IHCC (non-EBV IHCC) cases were collected. These samples were used to evaluate genetic alterations, TIME composition, and PD-L1 expression through ion AmpliSeq comprehensive cancer panel, PanCancer immune profiling panel, immunohistochemistry, and immunofluorescence staining. The correlation between clinical outcomes and TIME components was analyzed in the two EBV-LELCC cases receiving anti-PD-1 treatment. RESULTS: The genetic mutations identified in EBV-LELCC were BARD1, CD19, CD79B, EPHA5, KDM5A, MUC6, MUC16, PTEN, RECQL4, TET1, and TNFAIP3. Both CD79B and TNFAIP3 mutations were involved in the NF-κB signaling pathway. PD-L1 was highly expressed in tumor-infiltrating immune cells, especially the T cells and macrophages. The TIME of EBV-LELCC displayed abundant immune cell infiltration with a stronger adaptive immune response. Increased Th1 cells, NK CD56dim cells, and M1 macrophages, decreased M2 macrophages, exhausted CD8 T cell infiltration, and increased T cell activation signatures in TIME were associated with longer survival. Two patients with metastatic EBV-LELCC had good disease control after anti-PD-1 antibody treatment. A significantly larger TIME component made EBV-LELCCs more sensitive to immune checkpoint blockade (ICB). CONCLUSION: A better understanding of the composition of TIME in EBV-LELCC is critical for predicting the clinical outcomes of ICB treatment.
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Neoplasias de los Conductos Biliares , Colangiocarcinoma , Infecciones por Virus de Epstein-Barr , Antígeno B7-H1 , Neoplasias de los Conductos Biliares/complicaciones , Conductos Biliares Intrahepáticos/patología , Colangiocarcinoma/complicaciones , Colangiocarcinoma/terapia , Infecciones por Virus de Epstein-Barr/complicaciones , Herpesvirus Humano 4/genética , Humanos , Inhibidores de Puntos de Control Inmunológico , Inmunoterapia , Oxigenasas de Función Mixta , FN-kappa B , Proteínas Proto-Oncogénicas , Proteína 2 de Unión a Retinoblastoma , Microambiente TumoralRESUMEN
OBJECTIVE: We aimed to explore the underlying mechanism of Tripterygium glycosides (TGs) in treating systemic lupus erythematosus (SLE) through network-pharmacology approach. METHODS: The protein targets of TGs' three active ingredients (triptolide, tripterine, and wilforlide) and SLE were identified by database search. Then, the intersection of the two groups was studied. The drug-target network between the active ingredients of TGs and the overlapping genes was constructed, visualized, and analyzed with Cytoscape software. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment were performed to analyze these genes. Finally, we validated our predictions of the potential targets through docking study. RESULTS: A total of 55 overlapping genes were discovered. Results suggested that the TGs' mechanism in SLE treatment was associated with heat shock protein family A member 5, heat shock protein family A member 8, eukaryotic translation elongation factor 1 alpha 1, and so forth with their related 4042 gene network, which regulated ribosome, spliceosome, viral carcinogenesis, Epstein-Barr virus infection signaling, and so forth. Molecular-docking analysis proved that hydrogen bonding was the main form of interaction. CONCLUSIONS: Our research provided the protein targets affected by TGs in SLE treatment. The key targets (CASP3, MAPK1, HIF1A, and so forth) involving 4042 proteins became the multitarget mechanism of TGs in SLE treatment.
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Medicamentos Herbarios Chinos , Infecciones por Virus de Epstein-Barr , Lupus Eritematoso Sistémico , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Glicósidos/uso terapéutico , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Herpesvirus Humano 4/genética , Humanos , Lupus Eritematoso Sistémico/tratamiento farmacológico , Lupus Eritematoso Sistémico/genética , Tripterygium/efectos de los fármacosRESUMEN
This article aims to investigate the role of Simiao Qingwen Baidu Decoction (traditional Chinese medicine) in Epstein-Barr virus (EBV)-induced infectious mononucleosis. Sprague Dawley rats were given Simiao Qingwen Baidu Decoction by gavage, and the medicated serum was collected. EBV-latent infected human Burkitt lymphomas Raji and EBV-transformed marmosets B lymphoblast cell B95-8 were treated with medicated serum. CCK8 assay and flow cytometry were performed to detect cell proliferation and apoptosis. Indirect immunofluorescence assay was performed to analyze EA or VCA positive expression. The copy-number of EBV-DNA and the gene expression were detected by quantitative PCR or quantitative real-time PCR. We found that the medicated serum inhibited proliferation of Raji and B95-8 cells, especially 10 %-medicated serum. The 10 %-medicated serum significantly suppressed EA expression in Raji cells and VCA expression in B95-8 cells. The expression of BZLF1, BRLF1, BMLF1 and EBNA-1 in Raji cells was significantly inhibited by 10 %-medicated serum. 10 %-medicated serum caused a decrease in the copy-number of EBV-DNA in Raji cells. In conclusion, our data imply that Simiao Qingwen Baidu Decoction represses the expression of EA and VCA, and EBV-DNA replication. Thus, our work suggests that Simiao Qingwen Baidu Decoction may play a vital role in anti-EBV.
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Antígenos Virales , Proteínas de la Cápside/antagonistas & inhibidores , Replicación del ADN/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Regulación Viral de la Expresión Génica , Herpesvirus Humano 4/efectos de los fármacos , Animales , Antígenos Virales/genética , Antígenos Virales/metabolismo , Callithrix , Proteínas de la Cápside/genética , Proteínas de la Cápside/metabolismo , Línea Celular Transformada , Línea Celular Tumoral , Replicación del ADN/fisiología , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/metabolismo , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Replicación Viral/efectos de los fármacos , Replicación Viral/fisiologíaRESUMEN
Gastric carcinoma with lymphoid stroma is an uncommon variant enriched for mutually exclusive Epstein-Barr virus (EBV) positivity and mismatch repair (MMR) deficiency. We performed this study to evaluate molecular alterations in this morphologically homogeneous subtype and compare them with 295 conventional gastric cancers analyzed in The Cancer Genome Atlas study. We identified 31 study cases and subjected them to in situ hybridization for EBV-encoded RNAs and assessment for MMR status. Immunostains for PD-L1, ß-catenin, and HER2 were performed; extracted DNA was sequenced with a Comprehensive Cancer Panel. Most study patients were older adult men with stage I or II disease (76%). Tumors were classified as EBV/MMR-proficient (MMR-P) (n=7), EBV/MMR deficient (n=12), and EBV/MMR-P (n=12). EBV/MMR-P tumors were usually located in the proximal stomach (83%) and showed heterogenous growth patterns with glandular differentiation (83%). Tumors in all groups showed numerous tumor infiltrating lymphocytes and PD-L1 expression, infrequent nuclear ß-catenin accumulation (10%), and lacked both membranous HER2 staining and HER2 amplification. EBV/MMR-deficient tumors showed significantly higher tumor mutation burden (P=0.001) and KRAS alterations (56%) compared with EBV/MMR-P tumors (9%, P=0.05). TP53 variants were more common among EBV/MMR-P tumors (82%) compared with EBV/MMR proficient (0%, P=0.01) and EBV/MMR-deficient (11%, P<0.01) tumors. Alterations in KRAS, ARID1A, PIK3CA, and TP53 followed similar patterns of distribution compared with The Cancer Genome Atlas dataset. We conclude that gastric carcinomas with lymphoid stroma show a spectrum of molecular changes and frequent PD-L1 expression, raising the possibility that this subgroup of tumors may be susceptible to checkpoint inhibitors and/or agents that target receptor tyrosine kinase-mediated signaling.
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Biomarcadores de Tumor , Carcinoma/diagnóstico , Linfocitos Infiltrantes de Tumor , Neoplasias Gástricas/diagnóstico , Células del Estroma , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/genética , Biopsia , Carcinoma/química , Carcinoma/genética , Carcinoma/patología , Reparación de la Incompatibilidad de ADN , Femenino , Predisposición Genética a la Enfermedad , Herpesvirus Humano 4/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Inmunohistoquímica , Hibridación in Situ , Linfocitos Infiltrantes de Tumor/química , Linfocitos Infiltrantes de Tumor/patología , Masculino , Inestabilidad de Microsatélites , Persona de Mediana Edad , Mutación , Fenotipo , ARN Viral/genética , Estudios Retrospectivos , Neoplasias Gástricas/química , Neoplasias Gástricas/genética , Neoplasias Gástricas/patología , Células del Estroma/química , Células del Estroma/patologíaRESUMEN
PURPOSE: To assess the prognostic value of neutrophil-to-lymphocyte ratio (NLR) in patients with International Union Against Cancer (UICC)-staged III/IVA,B nasopharyngeal carcinoma (NPC), who were enrolled into two randomised controlled trials of concurrent/adjuvant chemotherapy when added to radiotherapy (SQNP01), and induction chemotherapy when added to chemoradiotherapy (NCC0901). MATERIAL AND METHODS: A post hoc analysis of pooled cohorts from SQNP01 (N = 221) and NCC0901 (N = 172) was performed. We employed a threshold of pre-treatment NLR = 3.0 (median) to stratify patients. Survival outcomes were compared using log-rank test. Multivariable Cox regression analyses were performed to assess association between NLR and overall survival (OS), disease-free survival (DFS), distant metastasis-free survival (DMFS), and locoregional recurrence-free survival (LRFS). RESULTS: High NLR (≥3.0) was associated with advanced T-status (p = 0.002), N-status (p = 0.002), overall UICC stage (p = 0.004), and high pre-treatment Epstein-Barr virus DNA titre (p = 0.001). High NLR was not associated with OS (0.94 [0.67-1.32], p = 0.7), DFS (0.98 [0.73-1.33], p = 0.9), DMFS (1.02 [0.66-1.57], p = 0.9), and LRFS (1.37 [0.84-2.22], p = 0.2) on univariable and multivariable analyses, while conventional clinical indices (T-status, N-status, and overall UICC stage) were prognostic of clinical outcomes. High NLR also did not predict for a treatment effect with the experimental arms in both trials. CONCLUSION: Our pooled analyses that were confined to a homogenous patient population of locally advanced NPC do not suggest that NLR adds prognostic value to conventional clinical indices in identifying patients with unfavourable disease.
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Carcinoma/sangre , Linfocitos , Neoplasias Nasofaríngeas/sangre , Neutrófilos , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carboplatino/administración & dosificación , Carcinoma/patología , Carcinoma/terapia , Carcinoma/virología , Cisplatino/administración & dosificación , ADN Viral/sangre , Desoxicitidina/administración & dosificación , Desoxicitidina/análogos & derivados , Supervivencia sin Enfermedad , Infecciones por Virus de Epstein-Barr/sangre , Femenino , Fluorouracilo/administración & dosificación , Herpesvirus Humano 4/genética , Humanos , Recuento de Leucocitos , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patología , Neoplasias Nasofaríngeas/terapia , Neoplasias Nasofaríngeas/virología , Estadificación de Neoplasias , Pronóstico , Modelos de Riesgos Proporcionales , Radioterapia , Ensayos Clínicos Controlados Aleatorios como Asunto , Singapur , Tasa de Supervivencia , GemcitabinaRESUMEN
This paper describes methodology for direct and indirect detections of a specific oligonucleotide for Epstein-Barr virus (EBV) using electrochemical techniques. The sequence of oligonucleotide probe (EBV1) revealed a high sequence identity (100%) with the EBV genome. For the development of the genosensor, EBV1 was grafted to the platform sensitized with poly(4-aminothiophenol). After that, the hybridization reaction was carried out with the complementary target (EBV2) on the modified electrode surface using ethidium bromide as DNA intercalator. The oxidation peak currents of ethidium bromide increased linearly with the values of the concentration of the complementary sequences in the range from 3.78 to 756 µmol·L⻹. In nonstringent experimental conditions, this genosensor can detect 17.32 nmol·L⻹ (three independent experiments) of oligonucleotide target, discriminating between complementary and non-complementary oligonucleotides, as well as differentiating one-base mismatch, as required for detection of genetic diseases caused by point mutations. The biosensor also displayed high specificity to the EBV target with elimination of interference from mix (alanine, glucose, uric acid, ascorbic acid, bovine serum albumin (BSA), glutamate and glycine) and good stability (120 days). In addition, it was possible to observe differences between hybridized and non-hybridized surfaces through atomic force microscopy.
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Biomarcadores/análisis , Técnicas Electroquímicas , Herpesvirus Humano 4/metabolismo , Polímeros/química , Compuestos de Anilina/química , Técnicas Biosensibles , Complejos de Coordinación/química , ADN Viral/metabolismo , Electrodos , Etidio/química , Herpesvirus Humano 4/genética , Humanos , Hibridación de Ácido Nucleico , Oligonucleótidos/química , Oligonucleótidos/metabolismo , Compuestos de Sulfhidrilo/químicaRESUMEN
PURPOSE: To investigate the effects of adjuvant chemotherapy in nasopharyngeal carcinoma (NPC) patients with persistently detectable plasma Epstein-Barr virus DNA (pEBV DNA) after curative radiation therapy plus induction/concurrent chemotherapy. METHODS AND MATERIALS: The study population consisted of 625 NPC patients with available pEBV DNA levels before and after treatment. Eighty-five patients with persistently detectable pEBV DNA after 1 week of completing radiation therapy were eligible for this retrospective study. Of the 85 patients, 33 were administered adjuvant chemotherapy consisting of oral tegafur-uracil (2 capsules twice daily) for 12 months with (n=4) or without (n=29) preceding intravenous chemotherapy of mitomycin-C, epirubicin, and cisplatin. The remaining 52 patients who did not receive adjuvant chemotherapy served as the control group. RESULTS: Baseline patient characteristics at diagnosis (age, sex, pathologic type, performance status, T classification, N classification, and overall stage), as well as previous treatment modality, were comparable in both arms. After a median follow-up of 70 months for surviving patients, 45.5% (15 of 33 patients) with adjuvant chemotherapy and 71.2% (37 of 52 patients) without adjuvant chemotherapy experienced tumor relapses (P=.0323). There were a significant reduction in distant failure (P=.0034) but not in local or regional recurrence. The 5-year overall survival rate was 71.6% for patients with adjuvant chemotherapy and 28.7% for patients without adjuvant chemotherapy (hazard ratio 0.27; 95% confidence interval 0.17-0.55; P<.0001). CONCLUSIONS: Our retrospective data showed that adjuvant chemotherapy can reduce distant failure and improve overall survival in NPC patients with persistently detectable pEBV DNA after curative radiation therapy plus induction/concurrent chemotherapy.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , ADN Viral/sangre , Herpesvirus Humano 4/genética , Neoplasias Nasofaríngeas/tratamiento farmacológico , Neoplasias Nasofaríngeas/virología , Adulto , Biomarcadores/sangre , Quimioterapia Adyuvante/métodos , Quimioterapia Adyuvante/mortalidad , Cisplatino/administración & dosificación , Combinación de Medicamentos , Epirrubicina/administración & dosificación , Femenino , Fluorouracilo/administración & dosificación , Humanos , Quimioterapia de Inducción/métodos , Leucovorina/administración & dosificación , Masculino , Persona de Mediana Edad , Mitomicina/administración & dosificación , Neoplasias Nasofaríngeas/mortalidad , Neoplasias Nasofaríngeas/radioterapia , Recurrencia Local de Neoplasia/mortalidad , Recurrencia Local de Neoplasia/virología , Radioterapia Conformacional/métodos , Estudios Retrospectivos , Tegafur/uso terapéutico , Resultado del Tratamiento , Uracilo/uso terapéuticoRESUMEN
Epstein--Barr virus (EBV) is a human virus with oncogenic potentials that is implicated in various human diseases and malignancies. In this study, the modulator activity of the potent herbal extract drug thymoquinone on EBV was assessed in vitro. Thymoquinone was tested for cytotoxicity on human cells of lymphoblastoid cells, Raji Burkitt's lymphoma, DG-75 Burkitt's lymphoma, peripheral blood mononuclear cells, and periodontal ligament fibroblast. Apoptosis induction was analyzed via TUNEL assay and activity studies of caspase-3. The effect of thymoquinone on EBV gene expression was determined using real-time polymerase chain reaction. We report here, for the first time, a promising selective inhibitory affect of thymoquinone on EBV-infected B cell lines in vitro, compared with lower activity on EBV negative B cell line and very low toxicity on human peripheral blood mononuclear cells and periodontal ligament fibroblasts. Moreover, the drug was found to efficiently suppress the RNA expression of EBNA2, LMP1, and EBNA1 genes. Specifically, EBNA2 expression levels were the most affected indicating that this gene might have a major contribution to thymoquinone potency against EBV infected cells. Overall, our results suggest that thymoquinone has the potential to suppress the growth of EBV-infected B cells efficiently.
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Linfocitos B/efectos de los fármacos , Linfocitos B/virología , Benzoquinonas/farmacología , Supervivencia Celular/efectos de los fármacos , Regulación Viral de la Expresión Génica/efectos de los fármacos , Herpesvirus Humano 4/efectos de los fármacos , Herpesvirus Humano 4/genética , Linfoma de Burkitt/tratamiento farmacológico , Linfoma de Burkitt/genética , Linfoma de Burkitt/virología , Caspasa 3/genética , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/genética , Infecciones por Virus de Epstein-Barr/tratamiento farmacológico , Infecciones por Virus de Epstein-Barr/genética , Infecciones por Virus de Epstein-Barr/virología , Fibroblastos/efectos de los fármacos , Fibroblastos/virología , Expresión Génica/efectos de los fármacos , Medicina de Hierbas/métodos , Herpesvirus Humano 4/metabolismo , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/virología , Células Tumorales Cultivadas , Proteínas Virales/genéticaRESUMEN
Burkitt lymphoma (BL) has been reported to be strongly associated with Epstein-Barr virus (EBV) infection. The fact that EBV is generally present in cancer cells but rarely found in healthy cells represents an opportunity for targeted cancer therapy. One approach is to activate the lytic replication cycle of the latent EBV. Nuclear factor (NF)-κB is thought to play an essential role in EBV lytic infection. Elevated NF-κB levels inhibit EBV lytic replication. Parthenolide (PN) is a sesquiterpene lactone found in medicinal plants, particularly in feverfew (Tanacetum parthenium). The aim of the present study was to analyze the effect of PN on the survival of Raji EBV-positive lymphoma cells. Raji cells were treated with 0, 4 or 6 µmol/l PN for 48 h. MTT assay and western blot analysis were performed to evaluate the findings. Results showd that PN suppressed the growth of the EBV-positive BL cell line, Raji, and activated the transcription of BZLF1 and BRLF1 by inhibiting NF-κB activity. Most notably, when PN was used in combination with ganciclovir (GCV), the cytotoxic effect of PN was amplified. These data suggest that the induction of lytic EBV infection with PN in combination with GCV may be a viraltargeted therapy for EBV-associated BL.
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Antivirales/toxicidad , Apoptosis/efectos de los fármacos , Herpesvirus Humano 4/metabolismo , Sesquiterpenos/toxicidad , Antivirales/química , Linfoma de Burkitt/etiología , Linfoma de Burkitt/metabolismo , Linfoma de Burkitt/virología , Línea Celular Tumoral , Infecciones por Virus de Epstein-Barr/complicaciones , Infecciones por Virus de Epstein-Barr/metabolismo , Infecciones por Virus de Epstein-Barr/virología , Ganciclovir/toxicidad , Herpesvirus Humano 4/genética , Humanos , Proteínas Inmediatas-Precoces/genética , Proteínas Inmediatas-Precoces/metabolismo , FN-kappa B/metabolismo , Plantas Medicinales/química , Sesquiterpenos/química , Tanacetum parthenium/química , Transactivadores/genética , Transactivadores/metabolismoRESUMEN
Epstein-Barr virus (EBV) may cause a variety of virus-associated diseases, but no antiviral agents have yet been developed against this virus. Animal models are thus indispensable for the pathological analysis of EBV-related infections and the elucidation of therapeutic methods. To establish a model system for the study of EBV infection, we tested the ability of B95-8 virus and recombinant EBV expressing enhanced green fluorescent protein (EGFP) to replicate in human lymphoid tissue. Human tonsil tissues that had been surgically removed during routine tonsillectomy were sectioned into small blocks and placed on top of collagen sponge gels in culture medium at the air-interface, then a cell-free viral suspension was directly applied to the top of each tissue block. Increasing levels of EBV DNA in culture medium were observed after 12-15 days through 24 days post-infection in tissue models infected with B95-8 and EGFP-EBV. Expression levels of eight EBV-associated genes in cells collected from culture medium were increased during culture. EBV-encoded small RNA-positive cells were detected in the interfollicular areas in paraffin-embedded sections. Flow cytometric analyses revealed that most EGFP(+) cells were CD3(-) CD56(-) CD19(+) HLA-DR(+), and represented both naïve (immunoglobulin D(+)) and memory (CD27(+)) B cells. Moreover, EBV replication in this model was suppressed by acyclovir treatment in a dose-dependent manner. These data suggest that this model has potential for use in the pathological analysis of local tissues at the time of primary infection, as well as for screening novel antiviral agents.
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Técnicas de Cultivo de Célula/métodos , Herpesvirus Humano 4/fisiología , Tonsila Palatina/citología , Tonsila Palatina/virología , Replicación Viral , Aciclovir/farmacología , Antivirales/farmacología , Separación Celular , Evaluación Preclínica de Medicamentos , Proteínas Fluorescentes Verdes/genética , Herpesvirus Humano 4/efectos de los fármacos , Herpesvirus Humano 4/genética , Humanos , Linfocitos/citología , Linfocitos/efectos de los fármacos , Linfocitos/virología , Tonsila Palatina/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
Epstein-Barr virus (EBV) expresses two transcription factors, Rta and Zta, during the immediate-early stage of the lytic cycle to activate the transcription of early and late genes. This study finds that 0.31 mM protoapigenone from Thelypteris torresiana (Gaud.) inhibits the expression of EBV lytic proteins, including Rta, Zta, EA-D and VCA, in P3HR1 cells after lytic induction with 12-O-tetradecanoylphorbol-13-acetate and sodium butyrate. The lack of expression of EBV lytic proteins after protoapigenone treatment is attributed to the inhibition of the transactivation function of Zta because protoapigenone reduces the transactivation activity of Zta and Gal4-Zta, which contains the transactivation domain of Zta fused with Gal4. In contrast, protoapigenone does not affect the ability of Rta to activate a promoter that contains an Rta-response element, showing that the inhibition is unrelated to Rta. Furthermore, in a lactate dehydrogenase assay, protoapigenone is not toxic to P3HR1 cells at the concentrations that inhibit the function of Zta, showing that protoapigenone is valuable for studying the function of Zta and preventing EBV lytic proliferation.
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Ciclohexanonas/farmacología , Regulación hacia Abajo/efectos de los fármacos , Helechos/química , Flavonas/farmacología , Herpesvirus Humano 4/fisiología , Extractos Vegetales/farmacología , Línea Celular , Infecciones por Virus de Epstein-Barr/tratamiento farmacológico , Infecciones por Virus de Epstein-Barr/virología , Regulación Viral de la Expresión Génica/efectos de los fármacos , Herpesvirus Humano 4/genética , Humanos , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
OBJECTIVE: To evaluate the clinical efficacy and safety of Chinese drugs for the treatment of children's infectious mononucleosis (CIM). METHODS: Sixty CIM patients were assigned into the treated group and the control group, patients in the treated group were administered with Chinese herbal decoction, and those in the control group were treated with intravenous dripping of ganciclovir 10 mg/kg per day, for a treatment course of 14 days. RESULTS: The total effective rate was 96.0% in the treated group and 97.1% in the control group, showing insignificant difference between groups. The efficacy in the treated group was superior to that in the control group on the fever clearance time (3.0+/-1.5 days vs 4.9+/-3.9 days ) and the disappearance time of cervical lymph node swelling (0.8+/-1.0 score vs 1.5+/-1.2 score), showing statistical significance (all P<0.05). T-cell subsets were markedly improved in both groups after treatment. Adverse reaction occurred in four cases of the control group. CONCLUSION: Using Chinese herbs for clearing heat, removing toxin, activating blood circulation, and dissolving stasis is effective and safe for the treatment of CIM. It can effectively improve the clinical symptoms and shows a certain effect on immune regulation.
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Mononucleosis Infecciosa/tratamiento farmacológico , Medicina Tradicional China , Antígenos CD/inmunología , Niño , Herpesvirus Humano 4/genética , Humanos , Mononucleosis Infecciosa/inmunología , Reacción en Cadena de la PolimerasaRESUMEN
This paper reports a simple DNA sensor having a detection limit of about 24 oligonucleotides and that operates without the need for PCR amplification. The sensor platform is based on an interdigitated array (IDA) of electrodes. The electrodes are modified with DNA capture probes, which are complementary to an analog for the Epstein-Barr genome, and then exposed to an alkaline phosphatase-labeled target. The enzyme catalyzes the formation of L-ascorbic acid, which reduces Ag(+) in solution to yield conductive Ag filaments that span the gap between the electrodes of the IDA. Resistance measurements, made with an inexpensive, hand-held multimeter, signal the presence of the target. The sensor response is insensitive to the presence of a large excess of non-complementary DNA sequences.
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Técnicas Biosensibles/instrumentación , Genoma Viral , Herpesvirus Humano 4/genética , Plata/química , Fosfatasa Alcalina/química , Fosfatasa Alcalina/metabolismo , Secuencia de Bases , Técnicas Biosensibles/métodos , Sondas de ADN/genética , Sondas de ADN/metabolismo , Electrodos , Herpesvirus Humano 4/aislamiento & purificación , Análisis de Secuencia por Matrices de Oligonucleótidos , Plata/metabolismoRESUMEN
BACKGROUND: Epstein-Barr virus (EBV) has been shown to be associated with gastric cancer. However, inconsistent findings have been reported regarding the distribution of EBV infected cells (in normal gastric epithelium vs. intestinal metaplastic cells vs. in neoplastic cells) and the characteristics of EBV-associated gastric cancer. Lymph node positive EBV-associated gastric cancer has not been systematically studied. The aims of this study were to evaluate EBV-associated gastric cancer, to assess the distribution of EBV infected cells including all positive lymph nodes, and to define the characteristics of EBV-associated gastric cancer. DESIGN: The study included primary gastric cancer patients who underwent surgical resection with no preoperative treatment at M.D. Anderson Cancer Center between 1987 and 2006. Formalin-fixed paraffin-embedded tissue from these resection specimens were assessed for EBV by in situ hybridization, the gold standard for EBV detection in tissue. EBV status was analyzed along with clinicopathologic parameters including age, gender, tumor type, lymph node status, and pathologic stage of the tumor. RESULTS: Among 235 patients, 12 had intranuclear expression of EBV. EBV staining was seen only in tumor cells and no detectable EBV was observed in normal gastric mucosa, intestinal metaplasia or stromal cells. Eight of 12 patients with EBV-associated gastric cancer had regional lymph node metastasis. Of note, metastatic tumor cells in all of the involved lymph nodes of these 8 cases contained EBV. The epidemiologic data showed 11 of the 12 patients with EBV-associated gastric cancer were men, ranging in age from 54 to 78 years (mean age, 60 years; median age, 62.1 years). The age distribution for non-EBV associated gastric cancer patients ranged from 21 to 93 years (mean age, 67 years; median age, 66.4 years). CONCLUSION: Our study demonstrated that EBV is present exclusively in gastric cancer cells. The detection of EBV in tumor cells in all of the lymph nodes involved with metastatic gastric carcinoma suggests simultaneous replication of EBV and tumor cells. The predominantly male gender and relatively younger age observed for the EBV-infected gastric cancer cases suggest an association between this disease and other factors, such as life style.
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Adenocarcinoma/virología , Infecciones por Virus de Epstein-Barr/patología , Herpesvirus Humano 4/aislamiento & purificación , Neoplasias Gástricas/virología , Infecciones Tumorales por Virus/patología , Adenocarcinoma/patología , Adenocarcinoma/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Infecciones por Virus de Epstein-Barr/virología , Femenino , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Mucosa Gástrica/virología , Herpesvirus Humano 4/genética , Humanos , Inmunohistoquímica , Hibridación in Situ , Masculino , Persona de Mediana Edad , ARN Viral/biosíntesis , Estudios Retrospectivos , Factores de Riesgo , Neoplasias Gástricas/patología , Neoplasias Gástricas/cirugía , Infecciones Tumorales por Virus/virología , Estados Unidos , Proteínas de la Matriz Viral/biosíntesis , Adulto JovenRESUMEN
Andrographis paniculata NEES is a medicinal plant that is commonly used in Asia. This work demonstrates that 25 microg/ml of ethanolic extract from A. paniculata (EEAP) and 5 microg/ml of andrographolide, a bioactive compound in EEAP, effectively inhibit the expression of Epstein-Barr virus (EBV) lytic proteins, Rta, Zta and EA-D, during the viral lytic cycle in P3HR1 cells. Transient transfection analysis revealed that the lack of expression of Rta, Zta and EA-D is caused by the inhibition of the transcription of BRLF1 and BZLF1, two EBV immediate-early genes that encode Rta and Zta, respectively. This study finds that the inhibition prevents the virus from producing mature viral particles. Meanwhile, andrographolide is not toxic to P3HR1 cells when the concentration is below 5 microg/ml, indicating that the compound is potentially useful as an anti-EBV drug.
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Antivirales/farmacología , Diterpenos/farmacología , Herpesvirus Humano 4/efectos de los fármacos , Proteínas Virales , Virión/efectos de los fármacos , Andrographis/química , Antivirales/aislamiento & purificación , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Diterpenos/aislamiento & purificación , Citometría de Flujo , Regulación Viral de la Expresión Génica/efectos de los fármacos , Genes Inmediatos-Precoces/genética , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/fisiología , Humanos , Componentes Aéreos de las Plantas/química , Transcripción Genética/efectos de los fármacos , Proteínas Virales/biosíntesis , Proteínas Virales/genética , Virión/genética , Virión/fisiología , Latencia del Virus/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
Epstein-Barr virus (EBV) causes infectious mononucleosis and is associated with various malignancies, including Burkitt's lymphoma and nasopharyngeal carcinoma. Like all herpesviruses, the EBV life cycle alternates between latency and lytic replication. During latency, the viral genome is largely silenced by host-driven methylation of CpG motifs and, in the switch to the lytic cycle, this epigenetic silencing is overturned. A key event is the activation of the viral BRLF1 gene by the immediate-early protein Zta. Zta is a bZIP transcription factor that preferentially binds to specific response elements (ZREs) in the BRLF1 promoter (Rp) when these elements are methylated. Zta's ability to trigger lytic cycle activation is severely compromised when a cysteine residue in its bZIP domain is mutated to serine (C189S), but the molecular basis for this effect is unknown. Here we show that the C189S mutant is defective for activating Rp in a Burkitt's lymphoma cell line. The mutant is compromised both in vitro and in vivo for binding two methylated ZREs in Rp (ZRE2 and ZRE3), although the effect is striking only for ZRE3. Molecular modeling of Zta bound to methylated ZRE3, together with biochemical data, indicate that C189 directly contacts one of the two methyl cytosines within a specific CpG motif. The motif's second methyl cytosine (on the complementary DNA strand) is predicted to contact S186, a residue known to regulate methyl-ZRE recognition. Our results suggest that C189 regulates the enhanced interaction of Zta with methylated DNA in overturning the epigenetic control of viral latency. As C189 is conserved in many bZIP proteins, the selectivity of Zta for methylated DNA may be a paradigm for a more general phenomenon.
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Metilación de ADN , Proteínas de Unión al ADN/genética , Epigénesis Genética , Regulación Viral de la Expresión Génica , Silenciador del Gen , Herpesvirus Humano 4/genética , Transactivadores/genética , Proteínas Virales/genética , Secuencia de Bases , Línea Celular Tumoral , Cisteína/metabolismo , Proteínas de Unión al ADN/metabolismo , Genoma Viral , Herpesvirus Humano 4/metabolismo , Herpesvirus Humano 4/patogenicidad , Humanos , Datos de Secuencia Molecular , Mutación Missense , Elementos de Respuesta/genética , Serina/metabolismo , Transactivadores/metabolismo , Proteínas Virales/metabolismoRESUMEN
To improve transgene expression of a non-viral gene delivery system, an Epstein-Barr virus (EBV)-based plasmid and cationic emulsion complex was prepared and evaluated. Cationic emulsion was formulated with castor oil, 3-N-(N',N'-dimethylaminoethane)-carbamoyl cholesterol (DC-Chol) and other co-emulsifiers. An EBV-based plasmid containing the two EBV components, origin of replication (oriP) and EBV nuclear antigen 1 (EBNA-1), was constructed. The physical characteristics of the emulsion and the emulsion/DNA complex were determined. After cells were transfected with cationic emulsion/EBV-based plasmid complex, transfection efficiency and expression pattern were evaluated using green fluorescent protein (GFP) as a reporter. The average particle size and zeta potential of the emulsion itself were 96 nm and + 17 mV, respectively. The emulsion showed stable size distribution up to at least one month. With an increase of emulsion to DNA ratio, zeta-potential increased from negative to positive and the particle size decreased to 200-300 nm. The complex was stable against DNase I digestion and showed comparable transfection efficiency with Lipofectin for several tested cell lines. An enhanced and prolonged gene expression was achieved using EBV-based plasmid and cationic emulsion complex. Combining physically stable emulsion with self-replicating EBV-based plasmid may confer more effective gene expression.
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Carcinoma Hepatocelular/genética , ADN Viral/administración & dosificación , ADN Viral/genética , Sistemas de Liberación de Medicamentos/métodos , Regulación Neoplásica de la Expresión Génica/genética , Herpesvirus Humano 4/genética , Transfección/métodos , Aceite de Ricino/química , Cationes , Línea Celular Tumoral , Colesterol/química , Emulsiones , Humanos , Neoplasias Hepáticas/genéticaRESUMEN
Cytomegalovirus (CMV) is responsible for significant morbidity and mortality in immunocompromised patients undergoing allogeneic hematopoietic stem cell transplantation. The limitations of antiviral drugs and a better understanding of the cellular immune response to CMV has lead to the development of alternative therapies that restore host cellular immunity to CMV. Infusion of donor T lymphocytes results in variable protection against CMV but a high incidence of graft-versus-host disease in the allogeneic setting. To prevent this complication and further improve anti-CMV immune response, several groups have developed new approaches, such as the introduction of a suicide gene to control alloreactivity against the host or the selective activation of CMV-specific T cells by antigen-presenting cells expressing CMV antigens introduced by gene transfer. Depending on the target cells and the strategy chosen, adenovirus, retrovirus or poxviruses derived vectors are used for gene transfer. The protocols as well as the preclinical and clinical results obtained in the field of anti-CMV immunotherapy using gene transfer are reported and discussed.