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Objective:To analyze the expression of human leukocyte antigen G (HLA-G) in human T-cell leukemia virus type 1 (HTLV-1)-positive T cells, and to investigate its role in the occurrence and development of HTLV-1 infection.Methods:The expression of HLA-G in HTLV-1-positive T cell lines (MT2 and MT4) was detected by Western blot and real-time PCR. HLA-G gene in MT2 and MT4 cells was knocked down by siRNA, and the effects of HLA-G on the expression of HTLV-1 Tax and P19 at mRNA and protein levels were detected by Western blot and real-time PCR. Moreover, the changes in cytokine expression in MT2 and MT4 cells were monitored at RNA level after HLA-G gene silencing. The proliferation ability of MT2 and MT4 cells was analyzed by CCK8. Signal transducer and activator of transcription 3 (STAT3) pathway-related proteins were detected by Western blot.Results:Compared with HTLV-1-negative T cells (Jurkat and MOLT4), the expression of HLA-G increased significantly in MT2 and MT4 cells. After knocking down the HLA-G gene with siRNA in MT2 and MT4 cells, the expression of HTLV-1 Tax and P19 at mRNA and protein levels was decreased, and the expression of antiviral cytokines IFN-γ and TNF-α was increased. The proliferation of MT2 and MT4 cells and STAT3 phosphorylation in these cells were decreased.Conclusions:HTLV-1 could induce T cells to overexpress the immune tolerance molecule HLA-G. Silencing HLA-G gene in HTLV-1-positive T cells could promote the production of antiviral cytokines and reduce IL-6 expression and STAT3 phosphorylation, thereby effectively inhibiting the replication of HTLV-1.
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Objective:To investigate the changes of circulating tumor DNA (ctDNA), circulating B cell-specific Moloney leukemia virus insertion site 1 mRNA (Bmi-1 mRNA) and microRNA-21 (miR-21) before and after treatment with epidermal growth factor receptor (EGFR) monoclonal antibody in advanced colorectal cancer, and analyze their association with treatment response.Methods:The clinical data of 98 patients with advanced colorectal cancer from March 2019 to March 2021 in Yantai Yuhuangding Hospital were retrospectively analyzed. After treatment with cetuximab, complete remission was in 4 cases, partial remission in 26 cases, stable disease in 39 cases, and progressive disease in 29 cases. The patients with complete remission and partial remission were classified as remission group (30 cases), the stable disease and progressive disease were classified as non-remission group (68 cases). Before treatment and after 2 cycles of treatment, the plasma level of ctDNA was detected by high-throughput sequencing; the levels of Bmi-1mRNA and miR-21 were detected by real-time fluorescence quantitative polymerase chain reaction. Spearman correlation was used to analyze the relationship between ctDNA, Bmi-1mRNA, miR-21 and treatment responsiveness after 2 cycles of treatment; multivariate Logistic regression was used to analyze the independent risk factors affecting treatment responsiveness; receiver operating characteristic (ROC) curve was drawn to evaluate the value of ctDNA, Bmi-1mRNA and miR-21 in predicting remission after 2 cycles of treatment.Results:There were no significant differences in ctDNA, Bmi-1mRNA and miR-21 before treatment between 2 groups ( P>0.05); the ctDNA, Bmi-1mRNA and miR-21 after 2 cycles of treatment in remission group were significantly lower than those in non-remission group: (10.03 ± 3.32) μg/L vs. (15.33 ± 5.14) μg/L, 0.13 ± 0.04 vs. 0.19 ± 0.05 and 0.81 ± 0.26 vs. 1.08 ± 0.24, and there were statistical differences ( P<0.01). Spearman correlation analysis result showed that ctDNA, Bmi-1mRNA and miR-21 were negatively correlated with treatment response ( r = -0.500, -0.506 and -0.531; P<0.01). Multivariate Logistic regression analysis result showed that, after controlling for the number of distant metastatic organs and clinical stage, ctDNA, Bmi-1mRNA and miR-21 were still independent risk factors for treatment response in patients with advanced colorectal cancer ( OR = 3.342, 2.725 and 1.838; 95% CI 3.116 to 3.584, 2.647 to 2.805 and 1.768 to 1.911; P<0.01). ROC curve analysis result showed that the area under the curve (AUC) of ctDNA, Bmi-1mRNA combined with miR-21 after 2 cycles of treatment to predict the treatment response was the largest with 0.922. Conclusions:The changes of ctDNA, Bmi-1mRNA and miR-21 in patients with advanced colorectal cancer before and after treatment with EGFR monoclonal antibody are related to the treatment response. Combined detection is helpful for screening patients sensitive to EGFR-targeted therapy, and can provide reference for new targets of molecular intervention.
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Abstract Background: Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). This disease mainly affects cattle, causing severe economic losses to producers. Objective: To establish individual and herd seroprevalence and determine the risk factors associated with BLV seropositivity for dairy and dual-purpose cattle herds in Ecuador. Methods: A total of 2,668 serum samples from 386 herds were collected. A questionnaire, including variables related to cattle health, management and the environment was completed by each herd. A commercial blocking enzyme-linked immunosorbent assay (ELISA) test was used to determine seropositivity. A generalized estimating equation model (GEE) was developed to determine the factors associated with BLV seropositivity. Results: Individual seroprevalence of BLV infection in Ecuador was 17.3% (CI95% = 15.86-18.74%). Herd prevalence was 37.8% (CI95% = 33.0-42.6%), and intra-herd prevalence ranged between 12.5 and 100% (median: 37.5%). The risk factors associated with BLV seropositivity were artificial insemination (OR: 2,215; CI95% =1.402-3.501), concrete floors (OR: 2.178; CI95% = 1.217-3.889), presence of wild ruminants (OR: 2.998; CI95% = 1.788-5.027), and sampling season (wet; OR: 1.996; CI95% = 1.140-3.497). Conclusions: Results indicate that BLV is widespread in cattle herds in Ecuador. In addition, the study suggests that a control program to fight BLV infection should focus on controlling the risk factors identified.
Resumen Antecedentes: El virus de la leucosis bovina (BLV) es el principal agente etiológico causante de la leucosis enzoótica bovina (EBL). Esta enfermedad afecta a los bovinos causando grandes pérdidas económicas a los productores. Objetivo: Establecer la seroprevalencia y dispersión del BLV, así como los factores de riesgo asociados a la seropositividad en explotaciones lecheras y de doble propósito en Ecuador. Métodos: Se recolectó un total de 2.668 muestras de suero de 386 explotaciones. Se aplicó un cuestionario que incluyó variables relacionadas con la salud del hato, medidas de manejo, y características ambientales de cada explotación. Para los análisis serológicos se utilizó un test inmunológico ligado a enzimas (ELISA). Para definir los factores de riesgo asociados a la seropositividad a BLV se desarrolló un modelo utilizando ecuaciones de estimación generalizadas (GEE). Resultados: La seroprevalencia de BLV en Ecuador fue de 17,3% (IC95% = 15,86-18,74%). La dispersión fue de 37,8% (IC95%= 33,0-42,6%), y la prevalencia intra-hato alcanzó rangos entre 12,5-100% (media: 37,5%). Los factores de riesgo asociados a la seropositividad a BLV fueron: inseminación artificial (OR: 2,215; IC95% = 1,402-3,501), piso de concreto (OR: 2,178; IC95% = 1,217-3,889), presencia de rumiantes salvajes (OR: 2,998; IC95% = 1,788-5,027), y temporada de muestreo (húmeda; OR: 1,996; IC95% = 1,140-3,497). Conclusiones: Los resultados indican que el BLV se encuentra disperso en las explotaciones de Ecuador. Adicionalmente, se sugiere la implementación de un programa de control para la lucha contra el BLV, debiéndose considerar medidas que se enfoquen al control de los factores de riesgo identificados en esta investigación.
Resumo Antecedentes: O vírus da leucemia bovina (BLV) é o principal agente causador da leucose enzoótica bovina (EBL). Esta doença afeta o gado causando graves prejuízos econômicos aos produtores. Objetivo: Estabelecer a soroprevalência e dispersão do BLV, assim como os fatores de risco associados à soropositividade nas produções leiteiras e de duplo propósito no Equador. Métodos: Um total de 2.668 amostras de soro de 386 explorações foram coletadas. Foi aplicado um questionário que incluía variáveis relacionadas à saúde do rebanho, medidas de manejo e ambiente para cada exploração. Para a análise sorológica foi utilizado um teste imunológico sobre enzimas (ELISA) para determinação da soropositividade. Para definir os fatores de risco associados à soropositividade a BLV, foi utilizado um modelo de equações estimativas generalizadas (GEE). Resultados: A soroprevalência de BLVno Equador é de 17,3% (IC95% = 15,86-18,74%). La dispersão de 37,8% (IC95% = 33,0-42,6%), e a prevalência intra-rebanho alcançou entre 12,5-100% (media: 37,5%). Os fatores de risco associados à soropositividade a BLV foram inseminação artificial (OR: 2,215; IC95% = 1,402-3,501), chão de concreto (OR: 2,178; IC95% = 1,217-3,889), presença de ruminantes selvagens (OR: 2,998; IC95% = 1,788-5,027) e época da amostragem (úmida; OR: 1,996; IC95% = 1,140-3,497). Conclusões: Os resultados indicam que o BLV se encontra disseminado nas explorações no Equador. Adicionalmente, o estudo pode contribuir para a implementação de um programa de controle para a luta contra o BLV, devendo-se considerar ações de controle dos fatores de risco identificados nesta investigação.
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Resumen El virus de la leucemia felina (ViLeF) es una de las principales enfermedades retrovirales de la familia Felidae que causan la muerte de sus individuos; de ahí interés diagnóstico y preventivo para la salud animal. El propósito de este artículo es determinar la prevalencia de infección por el ViLeF por serodiagnóstico del antígeno p27, en cuatro municipios del sur del valle de Aburrá, Colombia, usando los registros de los centros de diagnóstico del área. Se realizó un estudio descriptivo, transversal y retrospectivo, entre los años 2013-2015, que incluyó la revisión de 1718 pruebas diagnósticas de felinos domésticos del área urbana de Medellín, Envigado, Sabaneta y Caldas, procedentes de los centros de diagnóstico clínico del valle de Aburrá. El diagnóstico de ViLeF se realizó en muestras de suero por el inmunoensayo comercial Elisa (Idexx Laboratories©, Snap Combo Plus®, Maine, EUA). Los datos se procesaron en Statgraphics Centurión XVy se realizaron las pruebas estadísticas de Ji2 y Tukey. Del total de muestras, 376 (21,89 %) fueron positivas a la presencia del antígeno p27 de ViLeF. La edad de infectados osciló entre los 2 a 36 meses, hubo una mayor prevalencia en raza doméstica de pelo corto (DPC) y en machos. El porcentaje la prevalencia de ViLeF en el estudio fue de 21,88 %, siendo de importancia epidemiológica en el sur del Valle de Aburrá, Antioquia, Colombia.
Abstract Feline Leukemia Virus (FeLV) is one of the main retrovirus diseases of the Felidae family causing the death to the subjects. Therefore, there is a diagnostic and preventive interest regarding the animal health. This article aims to determine the infection prevalence due to FeLV after a p27 antigen serodiagnostic test applied in four towns in the southern Valle del Aburrá, Colombia, using the records of the diagnostic centers in each area. A descriptive, cross-sectional and retrospective study was conducted for the period 2013-2015, in which 1718 diagnostic tests from home felines were reviewed. These cases were from the urban area of Medellín, Envigado, Sabaneta and Caldas making part in the clinical diagnostic centers of the Valle de Aburrá. The FeLV diagnosis was conducted in serum samples with commercial immunoessay ELISA (IDEXX Laboratories©, Snap Combo Plus®, Maine, EUA). Data were processed in Statgraphics Centurión XV and statistical tests Ji2 and Tukey were conducted. Out of the total samples, 376 (21.89 %) were positive to p27 antigen for FeLV. The infected animals were from 2 to 36 months old. There was a higher prevalence among home races with short hair (SHR) and males. The FeLV prevalence percentage in the study was 21.88%, a figure with epidemiological significance in the southern Valle de Aburrá, Antioquia Province, Colombia.
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Resumen Las enfermedades producidas por virus como sida y leucemia son altamente prevalentes en felinos domésticos, debido a su facilidad de transmisión, presentan signos clínicos similares a otras infecciones que pueden generar dificultades en el diagnóstico, por lo tanto, se deben analizar mediante pruebas de laboratorio; los exámenes disponibles en la actualidad presentan algunas desventajas, por ello se estandarizó una PCR múltiple en tiempo real con sondas Taqman que fue diseñada para detección de infección con el virus de inmunodeficiencia felina (VIF), el virus de leucemia felina (VLFe) o mixtas. Se calculó la sensibilidad (0,53/0,26) y la especificidad (0,46/0,74) para leucemia y sida respectivamente con la metodología de Broemeling que considera que la prueba de referencia contra la cual se compara no es un referente verdadero (Gold standard), según estos resultados se concluye que es necesario aumentar el tamaño de muestra; sin embargo, la PCR múltiple es una metodología muy sensible, fue validada de forma In silico y presentó resultados congruentes al realizarla in vivo.
Abstract Diseases such as Feline Immunodeficiency Virus (VIF) and Feline Leukemia Virus (VLFe) are highly prevalent in domestic cats, due to their ease of transmission. Clinical signs are similar to other infections causing difficulties in diagnosis, therefore, they should be analyzed by laboratory tests; Currently available tests have some disadvantages, a multiplex real-time PCR was standardized with Taqman probes, designed for detection of infection with VIF, VLFe or both. Sensitivity (0.53/0.26) and specificity (0.46/0.74) for VLFe and VIF respectively, were calculated with Broemeling's methodology, which considers that the reference test against which it is compared is not a Gold standard, according to these results it is necessary to increase the sample size; However, multiplex PCR is a very sensitive methodology, it was validated In silico form and presented congruent results when performed in vivo.
Resumo Doenças causadas por vírus como AIDS e leucemia são altamente prevalentes em gatos domésticos, devido a sua facilidade de transmissão com sinais clínicos semelhantes a outras infecções que podem causar dificuldades no diagnóstico, portanto devem ser analisados por exames laboratoriais; Os testes atualmente disponíveis têm algumas desvantagens, portanto uma PCR em tempo real múltipla foi padronizada com sondas Taqman para a detecção de infecção pelo vírus da imunodeficiência felina (FIV), vírus da leucemia felina (VLFe) ou misto . A sensibilidade (0,53 /0,26) e a especificidade (0,46/0,74) para leucemia e AIDS foram calculadas, respectivamente, com a metodologia de Broemeling, que considera que o teste de referência com o qual é comparado não é um referência verdadeira (teste de ouro), de acordo com esses resultados, conclui-se que é necessário aumentar o tamanho da amostra, entretanto, a PCR múltipla é uma metodologia muito sensível, foi validada na forma silico e apresentou resultados congruentes quando realizada in vivo.
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Objective To investigate the expression changes of provirus integration site 1 for moloney murine leukemia virus(Pim-1) gene in damaged neurons in vitro and related molecular basis of neurotrophic factors regulating Pim-1 expression and promoting the neurite regeneration of damaged neurons. Methods Neuro-2a(N-2a)cells were induced into neuron-like N-2a(N-2a-N) cells by retinoic acid,the proliferation of N-2a cells was inhibited by deferoxamine mesylate (DFO), and N-2a-N cells were injured by acrylamide. The N-2a-N cells were divided into normal control group, injury group, ciliary neurotrophic factors (CNTF) group and neuritin (Nrn1) group, with four samples in each group. The phenotype of N-2a cells and the expression of Pim-1 protein in N-2a cells were detected by immunofluorescence cytochemistry, and the expression of Pim-1 in each group was detected by Real-time PCR and Western blotting. Western blotting was used to detect the expression changes of relevant molecules involving in regulating activity of Pim-1, cells survival, apoptosis and axonal regeneration. Results Cell immunofluorescence showed that N-2a-N cells had neuronal phenotype to express β-Ⅲ tubulin and neurofilament-200, and Pim-1 protein was expressed in N-2a-N cells. N-2a cell proliferation was effectively inhibited by 50 μmol/ L DFO, and N-2a-N cell damage model was established by 1 mmol/ L acrylamide. Pim-1 gene expression showed a tendency of first decreasing, then increasing, and then decreasing after N-2aN cells were injured. Compared with the injury group, the proportion of the longest neurite in CNTF group and Nrn1 group increased significantly, the expressions of intracellular signal transducers extracellular regulated protein kinase 1/ 2 (ERK1/ 2), phosphorylated extracellular regulated protein kinase 1/ 2 (p-ERK1/ 2), signal transducers and activators of transcription 3(STAT3), phosphorylated signal transducers and activators of transcription 3 (p-STAT3), and Pim-1 were up-regulated, the expressions of apoptosis-related molecules cleaved Caspase-3 and Bax were down-regulated, the expression of anti-apoptosis-related molecule Bcl-2 was up-regulated, so the growth-associated protein 43 (GAP-43) protein involved neurite regeneration. Conclusion There is a need to repair damaged N-2a-N cells by overexpressing the Pim-1 gene. CNTF and Nrn1 can activate the ERK1/ 2 and STAT3 signaling pathways of damaged N-2a-N cells, and then up-regulate the expression of Pim-1 and GAP-43,and then promote cell neurite regeneration.
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Abstract Bovine leukemia virus (BLV) is an important cattle pathogen that causes major economic losses worldwide, especially in dairy farms. The use of animal models provides valuable insight into the pathogenesis of viral infections. Experimental infections of sheep have been conducted using blood from BLV-infected cattle, infectious BLV molecular clones or tumor-derived cells. The Fetal Lamb Kidney cell line, persistently infected with BLV (FLK-BLV), is one of the most commonly used long-term culture available for the permanent production of virus. FLK-BLV cells or the viral particles obtained from the cell-free culture supernatant could be used as a source of provirus or virus to experimentally infect sheep. In this report, we aimed to determine the minimum amount of FLK-BLV cells or cell-free supernatant containing BLV needed to produce infection in sheep. We also evaluated the amount of antibodies obtained from a naturally-infected cow required to neutralize this infection. We observed that both sheep experimentally inoculated with 5000 FLK-BLV cells became infected, as well as one of the sheep receiving 500 FLK-BLV cells. None of the animals inoculated with 50 FLK-BLV cells showed evidence of infection. The cell-free FLK-BLV supernatant proved to be infective in sheep up to a 1:1000 dilution. Specific BLV antibodies showed neutralizing activity as none of the sheep became infected. Conversely, the animals receiving a BLV-negative serum showed signs of BLV infection. These results contribute to the optimization of a sheep bioassay which could be useful to further characterize BLV infection.
Resumen El virus de la leucosis bovina (bovine leukemia virus [BLV]) es un importante agente patógeno del ganado que causa importantes pérdidas económicas en todo el mundo, especialmente en los rodeos lecheros. El uso de modelos animales proporciona información valiosa sobre la patogénesis de las infecciones virales. Se realizaron infecciones experimentales en ovejas usando sangre de bovinos infectados con BLV, clones moleculares de BLV infecciosos o células derivadas de tumores. La línea celular Fetal Lamb Kidney, persistentemente infectada con el BLV (FLK-BLV), es uno de los cultivos a largo plazo más utilizados para la producción permanente de virus. Las células FLK-BLV o las partículas virales obtenidas del sobrenadante del cultivo libre de células podrían usarse como fuente de provirus o de virus para infectar experimentalmente ovejas. En este trabajo, nuestro objetivo fue determinar la cantidad mínima de células FLK-BLV o de sobrenadante libre de células que contiene BLV necesaria para producir infección en ovejas. También evaluamos la cantidad de anticuerpos bovinos anti-BLV necesaria para neutralizar la infección. Observamos que las dos ovejas inoculadas experimentalmente con 5000 células FLK-BLV se infectaron, y que una de las dos ovejas que recibieron 500 células FLK-BLV se infectó. Ninguno de los animales inoculados con 50 células FLK-BLV mostró evidencia de infección. El sobrenadante FLK-BLV libre de células demostró ser infectivo en ovejas hasta la dilución 1:1000. Los anticuerpos BLV específicos mostraron actividad neutralizante, ya que ninguna de las ovejas se infectó. Por el contrario, los animales que recibieron un suero BLV negativo mostraron signos de infección por BLV. Estos resultados contribuyen a la optimización de un bioensayo en ovejas útil para caracterizar la infección por BLV.
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Animales , Bioensayo/veterinaria , Ovinos/inmunología , Leucosis Bovina Enzoótica/prevención & control , Virus de la Leucemia Bovina/patogenicidad , Infecciones por Deltaretrovirus/inmunología , Modelos AnimalesRESUMEN
Enzootic bovine leukosis (EBL) is an infectious disease of cosmopolitan distribution and chronic character caused by a virus of the Retroviridae family, bovine leukemia virus (BLV). The epidemiological situation of EBL in Brazil has motivated studies to improve its diagnosis, based on the recommended serological techniques: agar gel immunodiffusion (AGID) and enzyme-linked immunosorbent assay (ELISA). This study was designed to evaluate the use of imported ELISA for the detection of BLV in dairy herds raised in Pernambuco, Brazil, comparing it to AGID. Blood serum samples from 327 dairy cattle from the state of Pernambuco were tested to AGID and the imported commercial ELISA CHEKIT-Leucose-serum, produced by the IDEXX® laboratory for the diagnosis of EBL. Discarding 25 inconclusive samples from one or both tests, 302 samples were analyzed, being 24.1% positive (73/302) in the AGID and 45% (136/302) in the ELISA, which compared to the AGID, a technique considered standard, presented sensitivity of 98.6%, specificity of 72% and Kappa coefficient of 0.55. The lack of agreement in the diagnostic methods was probably due to the high sensitivity of the ELISA, which makes it possible to detect antibodies even in situations with low serum levels. Although AGID has been shown to be an efficient test so far, in more advanced stages of an EBL control and eradication program, with low prevalence rates, ELISA will present better performance, due to its higher sensitivity, avoiding the permanence of animals that spread the disease in the herds.(AU)
A leucose enzoótica bovina (LEB) é uma doença infecciosa de distribuição cosmopolita e caráter crônico causada por um vírus da família Retroviridae, o vírus da leucemia bovina (VLB). A situação epidemiológica da LEB no Brasil vem motivando estudos para o aprimoramento do seu diagnóstico, tendo como base as técnicas sorológicas recomendadas: imunodifusão em gel de ágar (IDGA) e Enzyme-Linked Immunoabsorbent Assay (ELISA). Este estudo teve como objetivo avaliar o uso de ELISA importado para a detecção do VLB em rebanhos leiteiros criados em Pernambuco, Brasil, comparando-o ao IDGA. Amostras de soro sanguíneo de 327 bovinos leiteiros do estado de Pernambuco foram testadas para IDGA e ELISA comercial importado CHEKIT-Leucose-serum, produzido pelo laboratório IDEXX® para o diagnóstico da LEB. Descartadas 25 amostras inconclusivas de um ou ambos os testes, foram analisadas 302 amostras, sendo 24,1% positivas (73/302) na IDGA e 45% (136/302) no ELISA, que em relação à IDGA, técnica considerada padrão, apresentou sensibilidade de 98,6%, especificidade de 72% e coeficiente Kappa de 0.55. A falta de concordância entre os métodos diagnósticos deveu-se, provavelmente, à elevada sensibilidade do ELISA, que possibilita detectar anticorpos mesmo em situações com baixos teores séricos. Apesar da IDGA se mostrar até o momento um teste eficiente, em etapas mais avançadas de um programa de controle e erradicação da LEB, com baixos índices de prevalência, o ELISA apresentará melhor desempenho, por possuir maior sensibilidade, evitando-se a permanência de animais disseminadores da doença nos rebanhos.â(AU)
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Animales , Bovinos , Pruebas Serológicas/métodos , Leucosis Bovina Enzoótica/diagnóstico , Virus de la Leucemia Bovina/inmunología , Bovinos , Ensayo de Inmunoadsorción Enzimática , Sensibilidad y Especificidad , Inmunodifusión/métodosRESUMEN
B lymphoma Moloney murine leukemia virus insertion region 1 (BMI1), a core member of polycomb repressive complex 1 (PRC1), has been intensely investigated in the field of cancer epigenetics for decades. Widely known as a critical regulator in cellular physiology, BMI1 is essential in self-renewal and differentiation in different lineages of stem cells. BMI1 also plays a significant role in cancer etiology for its involvement in pathological progress such as epithelial-mesenchymal transition (EMT) and cancer stem cell maintenance, propagation, and differentiation. Importantly, overexpression of BMI1 is predictive for drug resistance, tumor recurrence, and eventual therapy failure of various cancer subtypes, which renders the pharmacological targeting at BMI1 as a novel and promising therapeutic approach. The study on prostate cancer, a prevalent hormone-related cancer among men, has promoted enormous research advancements in cancer genetics and epigenetics. This review summarizes the role of BMI1 as an oncogenic and epigenetic regulator in tumor initiation, progression, and relapse of prostate cancer.
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Objective@#To investigate the effects of human adult T lymphoblastic leukemia virus typeⅠ (HTLV-1) infection on the production of reactive oxygen species (ROS) and mitochondrial damage in host cells.@*Methods@#A cell model of HTLV-1 infection was established by co-culturing HTLV-1-positive cell line MT2 with HeLa cells. ROS, mitochondrial membrane potential (MMP) and total mitochondria were detected using specific fluorescence probe labeling method. Cell apoptosis was detected by Annexin V-FITC/PI method. Western blot was performed to detect viral proteins Tax and p19, as well as mitochondrial proteins TIM23 and TOM20. After the treatment of MT2 cells with different concentrations of reverse transcription inhibitors (ZDV), relative viral loads were detected by quantitative real-time PCR and Western blot, and the mass of mitochondria was analyzed by flow cytometry.@*Results@#After co-culturing HeLa cells with MT2 cells for 24 h, the ROS level in host cells increased without obvious cell apoptosis, while the mitochondrial membrane potential, mitochondrial protein expression and total mitochondria decreased significantly. When the replication of HTLV-1 in MT2 cells was inhibited by ZDV, the ROS level and total mitochondria increased.@*Conclusions@#HTLV-1 infection can cause oxidative stress in host cells, resulting in mitochondrial damage. Autophagy might be activated to degrade mitochondrial damage and maintain cell homeostasis during the infection.
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Objective To investigate the effects of human adult T lymphoblastic leukemia virus typeⅠ (HTLV-1) infection on the production of reactive oxygen species (ROS) and mitochondrial damage in host cells. Methods A cell model of HTLV-1 infection was established by co-culturing HTLV-1-positive cell line MT2 with HeLa cells. ROS, mitochondrial membrane potential ( MMP) and total mitochondria were detected using specific fluorescence probe labeling method. Cell apoptosis was detected by Annexin V-FITC/PI method. Western blot was performed to detect viral proteins Tax and p19, as well as mitochondrial pro-teins TIM23 and TOM20. After the treatment of MT2 cells with different concentrations of reverse transcrip-tion inhibitors ( ZDV) , relative viral loads were detected by quantitative real-time PCR and Western blot, and the mass of mitochondria was analyzed by flow cytometry. Results After co-culturing HeLa cells with MT2 cells for 24 h, the ROS level in host cells increased without obvious cell apoptosis, while the mitochon-drial membrane potential, mitochondrial protein expression and total mitochondria decreased significantly. When the replication of HTLV-1 in MT2 cells was inhibited by ZDV, the ROS level and total mitochondria increased. Conclusions HTLV-1 infection can cause oxidative stress in host cells, resulting in mitochon-drial damage. Autophagy might be activated to degrade mitochondrial damage and maintain cell homeostasis during the infection.
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Objective To investigate whether cyclic GMP-AMP synthase ( cGAS ) , a cytosolic DNA sensor, could recognize the reverse transcription intermediate and induce the subsequent signaling path-way during the infection of human T cell leukemia virus type 1 ( HTLV-1 ) . Methods Biotin-labeled ssDNA90, a reverse transcription intermediate of HTLV-1, was transfected into HeLa cells and the interac-tion between it and cGAS was detected by co-immunoprecipitation experiments. HeLa cells were co-cultured with HTLV-1-positive MT2 cells and the interaction between cGAS and stimulator of interferon genes ( STING) was analyzed by co-immunoprecipitation experiments. The expression of STING in HeLa cells was silenced by siRNA. cGAS was transfected into the HeLa cells 24 h after the silencing and after 24 h, these cells were co-cultured with MT2 cells for another 24 h. Real-time PCR assay was used to measure the ex-pression of IFN-β, RANTES ( regulated upon activation, normal T-cell expressed, and secreted) , TNF-α, HTLV-1 protein Tax, p19 and HBZ. Immunoblot assay was performed to evaluate the phosphorylation of IRF3 and p65 in HeLa cells. Results cGAS interacted with ssDNA90. cGAS interacted with STING in the cytoplasm. In STING-silenced HeLa cells, cGAS transfection had no influence on the expression of IFN-β, RANTES , TNF-α, Tax , p19 or HBZ , nor did it affect the phosphorylation of IRF3 or p65 . Conclusions cGAS interacted with HTLV-1 RTI ssDNA90 and activated STING-dependent innate immune responses.
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Objective To determine the role of B cell specific moloney leukemia virus insert site 1 (Bmil) in hereditary gingival fibromatosis (HGF). Methods The HE staining was used to analyze the HGF and normal groups. The protein and mRNA of the Bmil, PCNA and caspase-3 in 2 groups were detected by immunohistochemistry and PCR, respectively. Results In HGF group, the gingival epithelial was incrassation, epithelial spikes was elongation, connective tissue was rich in fibroblast and collagen fibers, aless blood vessels and mild inflammatory hyperplasia. Bmil expression was higher (P < 0.05) and caspase-3 expression was lower (P < 0.05) in HGF group than in normal group. There was no difference of PCNA expression in the 2 groups (P> 0.05).Conclusion The Bmil might have a role in the pathogenesis of HGF by decreasing caspase-3 and caspase-3 mRNA.
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B lymphoma Moloney murine leukemia virus insertion region 1 (BMI1), a core member of polycomb repressive complex 1 (PRC1), has been intensely investigated in the field of cancer epigenetics for decades. Widely known as a critical regulator in cellular physiology, BMI1 is essential in self-renewal and differentiation in different lineages of stem cells. BMI1 also plays a significant role in cancer etiology for its involvement in pathological progress such as epithelial-mesenchymal transition (EMT) and cancer stem cell maintenance, propagation, and differentiation. Importantly, overexpression of BMI1 is predictive for drug resistance, tumor recurrence, and eventual therapy failure of various cancer subtypes, which renders the pharmacological targeting at BMI1 as a novel and promising therapeutic approach. The study on prostate cancer, a prevalent hormone-related cancer among men, has promoted enormous research advancements in cancer genetics and epigenetics. This review summarizes the role of BMI1 as an oncogenic and epigenetic regulator in tumor initiation, progression, and relapse of prostate cancer.
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Animales , Humanos , Masculino , Ratones , Regulación Neoplásica de la Expresión Génica , Linfoma de Células B/genética , Virus de la Leucemia Murina de Moloney/genética , Mutagénesis Insercional/genética , Complejo Represivo Polycomb 1/genética , Neoplasias de la Próstata/genéticaRESUMEN
RESUMEN El virus de la leucosis bovina (VLB) es un retrovirus que afecta principalmente el ganado lechero, reduciendo la producción de leche entre el 2,5 y 5%. La raza criolla colombiana Blanco Orejinegro (BON) es una raza rustica, bien adaptada, que ha mostrado resistencia in vitro a las infecciones ocasionadas por los virus de la fiebre aftosa y la estomatitis vesicular, así como las originadas por la bacteria Brucella abortus. El objetivo del presente estudio fue determinar si la raza BON y su cruce con Holstein son resistentes a la infección por el VLB. Se tomaron 124 muestras de sangre (59 Holstein, 40 BON y 25 BON x HOL) del mismo hato, se extrajo el DNA y se realizó una PCR-anidada correspondiente a una región del gen env de VLB. Se obtuvo un fragmento de 444 pb en los animales positivos. La prevalencia molecular del hato fue 33% para VLB. Se encontró diferencia significativa para infección por VLB entre los tres grupos raciales (p < 0,05). El porcentaje de infección fue del 55,9% para la raza Holstien, 5% para las vacas BON y 24% para el cruce BON x HOL; este último presentó una reducción en el porcentaje de infección del 32% respecto a la raza Holstein, lo cual puede ser atribuido a la presencia de genes de resistencia en la raza BON. Se comprobó que el nivel de infección es menor en vacas lecheras del cruce BON x HOL que en la raza lechera Holstein.
ABSTRACT The bovine leukemia virus (BLV) is a retrovirus that primarily affects dairy cattle, reducing milk production between 2.5 and 5%. The Colombian Blanco Orejinegro (BON) is a well-adapted, rustic, creole breed resistant to in vitro infections of Foot-and-mouth disease virus and vesicular stomatitis virus, as well as to Brucella abortus. This study aimed to determine if the crossing of BON and Holstein breeds is resistant to infection by BLV. Blood samples of 124 individuals (59 Holstein, 40 BON, and 25 BON x HOL) of the same herd were taken. The DNA was extracted, and a nested PCR was performed related to a region of the env gene of BLV. A fragment of 444 bp was obtained for positives animals. The molecular in-herd prevalence was 33% for BLV. A significant difference for BLV infection was found among the groups (p<0.05). The infection rate for the Holstein group was 55.9%, for BON cattle 5%, and for BON x HOL cattle 24%. The latter showed a reduction in the infection rate of 32% to the Holstein breed, which can be attributed to the presence of resistance genes in the BON breed. It was found that the level of infection is lower in BON x HOL cattle in contrast with Holstein dairy cows.
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A colângio-hepatite é considerada uma causa frequente de insuficiência hepática em gatos e é classificada em neutrofílica, linfocítica e esclerosante. Os objetivos deste estudo foram determinar a frequência de colângio-hepatite em gatos diagnosticados na Região Metropolitana de Porto Alegre, descrever seus aspectos anatomopatológicos e estabelecer uma associação com as infecções por Escherichia coli, vírus da imunodeficiência felina (FIV) e vírus da leucemia felina (FeLV). No período de janeiro de 2000 a julho de 2016 o Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul realizou 1915 necropsias de gatos, destes, 32 foram diagnosticados com colângio-hepatite, representando 1,7% dos casos. Destes, a colângio-hepatite linfocítica (CHL) foi diagnosticada em 68,7% (22/32), a neutrofílica (CHN) em 21,9% (7/32) e a esclerosante (CHE) com 9,4% (3/32). A idade variou de quatro meses a 16 anos, com a mediana de seis anos, acometendo predominantemente gatos sem raça definida. Somente na CHN observou-se predisposição por machos, verificado em 85,7% (6/7) dos casos. Enterite e pancreatite foram identificadas concomitantemente com a colângio-hepatite em 56,2% (18/32) dos casos, cada, e a formação de tríade foi identificada em 46,9% (15/32) dos gatos. Através da imuno-histoquímica, 68,2% (15/22) dos gatos com CHL, foram positivos para FIV, 40,9% (9/22) para FeLV e 31,8% (7/22) marcação para ambos os retrovírus. Na CHN, 85,7% (6/7) positivos para FIV, 57,1% (4/7) para FeLV e 42,8% (3/7) imunorreação para os dois retrovírus. Na CHE, 100% (3/3) dos casos apresentaram marcação para FeLV, 33,3% (1/3) para FIV e 33,3% (1/3) para ambos. Imunomarcação para E. coli foi observada em 27,3% (6/22) dos casos da CHL, 28,6% (2/7) da CHN e em 33,3% (1/3) da CHE. E. coli, Enterococcus sp. e Klebsiella pneumoniae foram os micro-organismos mais frequentes isolados no exame bacteriológico. A visualização da E. coli, através da IHQ no sistema hepatobiliar de gatos diagnosticados com colângio-hepatite associados à inflamação, sugere que a doença se desenvolveu secundariamente à infecção bacteriana ascendente.(AU)
Cholangiohepatitis is considered a frequent cause of liver failure in cats, and are classified as neutrophilic, lymphocytic and sclerosing. The aims of this study was to determine the frequency of cholangiohepatitis in cats diagnosed in the metropolitan region of Porto Alegre, to describe the anatomopathological aspects and to establish an association with Escherichia coli, feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). From January 2000 to July 2016, the Department of Veterinary Pathology of the Universidade Federal do Rio Grande do Sul performed 1915 cat necropsies, of which 32 were diagnosed with cholangiohepatitis, representing 1.7% of the cases. Of these, lymphocytic cholangiohepatitis (LCH) was diagnosed in 68.7% (22/32), neutrophilic (NCH) in 21.9% (7/v) and sclerosing cholangiohepatitis (SCH) with 9.4% (3/32) of the cases. In general, age ranged from four months to 16 years, with the median age of six years, and predominantly affected cats with mixed breed. Only in NCH demonstrated male predisposition, verified in 85.7% (6/7) of this cases. Enteritis and pancreatitis were identified concomitantly with cholangiohepatitis in 56.2% (18/32) cases, each, and triad formation was identified in 46.9% (15/32) of cats. In the immunohistochemistry, we observed that 68.2% (15/22) of the cats with LCH were positive for FIV, 40.9% (9/22) for FeLV, and 31.8% (7/22) positive for both retroviruses. In NCH, 85.7% (6/7) was positive for FIV, 57.1% (4/7) for FeLV, and 42.8% (3/7) immunoreactions for the two retroviruses. In SCH, 100% (3/3) of the cases presented FeLV marking, 33.3% (1/3) for FIV and 33.3% (1/3) for both. Immunostaining for E. coli was observed in 27.3% (6/22) of LCH, 28.6% (2/7) of NCH, and 33.3% (1/3) of SCH. E. coli, Enterococcus sp. and Klebsiella pneumoniae were the most frequently microorganisms isolated in the bacteriological examination. The visualization of E. coli by the immunohistochemistry in the hepatobiliary system of cats diagnosed with cholangiohepatitis suggests that the disease developed secondary to ascending bacterial infection.(AU)
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Animales , Gatos , Gatos/anomalías , Colangitis/diagnóstico , NoxasRESUMEN
Abstract Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp ® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.
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Animales , Bovinos , Ensayo de Inmunoadsorción Enzimática/métodos , Pruebas Serológicas/métodos , Leucosis Bovina Enzoótica/diagnóstico , Virus de la Leucemia Bovina/inmunología , Proteínas de la Cápside/inmunología , Anticuerpos Antivirales/sangre , Proteínas Recombinantes/análisis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Ensayo de Inmunoadsorción Enzimática/instrumentación , Sensibilidad y Especificidad , Leucosis Bovina Enzoótica/sangre , Leucosis Bovina Enzoótica/virología , Virus de la Leucemia Bovina/aislamiento & purificación , Virus de la Leucemia Bovina/genética , Proteínas de la Cápside/análisis , Proteínas de la Cápside/genéticaRESUMEN
Abstract Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp ® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.
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Bovine leukemia virus (BLV) is a member of Retroviridae family, genus Deltaretrovirus, and the main viral agent responsible for economic loses in dairy herds. Some studies have been carried out about BLV genotypes, and at least seven genotypes were found out in samples of different regions of the world. The objective of this study was to identify BLV samples from seropositive dairy cattle in Santa Catarina state, Brazil, using molecular techniques. Blood samples were collected (454) from dairy cattle from 31 different farms, and serology using agar gel immunodiffusion test (AGID) was performed. After that, 191 seropositive samples were submitted to DNA extraction, and in 77 samples the polymerase chain reaction (PCR) for amplification of a 440 bp fragment of the env gene was performed. Nineteen DNA samples were subjected to restriction fragment length polymorphism (RFLP) analysis by digestion of the PCR fragment by five restriction endonucleases - BamHI, HaeIII, Tru9I, TaqI, and MwoI. It was found 42% seropositive animals (191/454) and 68% positives of the farms (21/31). The PCR showed 80.5% (62/77) of animals positive. The RFLP analysis identified five different genotypes dispersed by Santa Catarina state, with the highest prevalence for genotype X (47.4%). Overall, our results identified the viral genotypes present in dairy cattle and the prevalence of new variants in representative farms from Santa Catarina state.(AU)
O bovine leukemia virus (BLV) é um membro da família Retroviridae, gênero Deltaretrovirus, e o principal agente viral causador de perdas econômicas em rebanhos leiteiros. Diversos estudos têm sido feitos sobre os genótipos de BLV, e foram encontrados pelo menos sete em amostras de diferentes partes do mundo. O objetivo deste estudo foi realizar a caracterização molecular de amostras de BLV de bovinos leiteiros soropositivos no estado de Santa Catarina. Foram coletadas 454 amostras de sangue de bovinos de 31 propriedades, e fez-se inicialmente a sorologia por meio do teste de imunodifusão em gel de ágar. Após a sorologia, 191 amostras soropositivas foram então submetidas à extração de DNA, e em 77 amostras se realizou a reação da polimerase em cadeia (PCR), para a amplificação de um fragmento de 440 pb do gene env. Dezenove amostras foram submetidas à análise do polimorfismo dos fragmentos de restrição por digestão do fragmento da PCR por cinco enzimas de restrição: BamHI, HaeIII, Tru9I, TaqI e MwoI. Os resultados obtidos na sorologia apontaram 42% de animais soropositivos (191/454) e 68% de propriedades positivas (21/31). Na PCR, 80,52% (62/77) dos animais apresentaram-se positivos. A análise do polimorfismo dos fragmentos de restrição identificou cinco genótipos circulantes no estado, e a maior prevalência foi observada no genótipo X (47,4%). Este estudo permite-nos conhecer alguns dos genótipos virais presentes em bovinos leiteiros do estado de Santa Catarina, bem como identificar a existência de novas variantes e sua prevalência atual, e os resultados são úteis para futuros estudos epidemiológicos.(AU)
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Bovinos , Pruebas Serológicas/métodos , Leucosis Bovina Enzoótica , Virus de la Leucemia Bovina , Leche , Reacción en Cadena de la Polimerasa/métodos , Agroindustria/economíaRESUMEN
Objective:To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors ofnasopharyngeal carcinoma cells.Methods:Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed,and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously.The integration of target genes were detected by PCR,the expressions of E-cad and Bmi-1 were detected by Western blot.The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay.T-he cell cycle and apoptosis were detected by flow cytometry.The cell migration and invasion were detected by Transwell assay.Results:The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells,the protein expression level of E-cad was up-regulated,and the protein expression level of Bmi-1 was down-regulated.The intervention of E-cad and Bmi-1 didn't affect the proliferation,cell cycle and apoptosis of CNE-2 cells,but it significantly inhibited the migration and invasion ability of CNE-2 cells.Furthermore,the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P＜0.01).Conclusion:The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro,which may lay the preliminary experimental basis for gene therapy of human cancer.