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1.
Virol Sin ; 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31828587

RESUMO

HIV-1 Rev is an accessory protein that plays a key role in nuclear exportation, stabilization, and translation of the viral mRNAs. Rev of HIV-1 clade BC often shows a truncation of 16 AAs due to a premature stop codon at residue 101. This stop codon presents the highest frequency in clade BC and the lowest frequency in clade B. In order to discover the potential biological effect of this truncation on Rev activity and virus replication of clade BC, we constructed Rev expression vectors of clade BC with or without 16 AAs within C-terminal separately, and replaced the stop codon by Q in a CRF07_BC infectious clone. We found that 16 AAs truncation had no effect on expression and activity of Rev in clade BC. Also, the mutation from the stop codon to Q had no effect on virus replication of clade BC. Next, to investigate the effect of this truncation on Rev activity and replication capacity of clade B, Rev expression vectors of clade B carrying or lacking 16 AAs in C-terminal were constructed respectively, and residue Q at position 101 within Rev was substituted by the stop codon in a clade B infectious clone. It was found that 16 AAs truncation significantly down-regulated Rev expression and impaired clade B Rev activity. Furthermore, a Q-to-stop codon substitution within Rev significantly reduced viral replication fitness of clade B. These results indicate that the premature stop codon at residue 101 within Rev exerts diverse impact on viral replication among different HIV-1 clades.

2.
Nat Commun ; 10(1): 2257, 2019 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-31113957

RESUMO

A major barrier to human immunodeficiency virus (HIV) cure is the existence of viral reservoirs that lead to viral rebound following discontinuation of antiretroviral therapy (ART). We postulate that enhancing cytotoxic T lymphocytes (CTL) targeting conserved envelope (Env) regions can eliminate HIV infected cells in latency. Here, we evaluate the use of adoptively transferred HIV vaccine-induced subtype C Env-specific CTLs in a macaque subtype B simian-human immunodeficiency virus (SHIV) model to determine whether plasma viremia can be controlled after ART interruption. We demonstrate that adoptive cellular therapy (ACT) using autologous Env-specific T cells augmented by therapeutic vaccination can suppress ART-free viral rebound in the SHIV model. Furthermore, phenotypic and functional characterization of adoptively transferred cells in ACT-responsive and nonresponsive animals support a critical role for cross-reactive central memory T cells in viremia control. Our study offers an approach to potentiate immunological suppression of HIV in the absence of antiviral drugs.

3.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 44(2): 201-208, 2019 Feb 28.
Artigo em Chinês | MEDLINE | ID: mdl-30837390

RESUMO

Articular cartilage lesions due to injury or other pathology are often difficult to heal, and the outcomes of the clinical treatment widely used today are far from satisfaction. Adipose-derived stem cells (ADSCs) are multipotent stem cells from adipose tissue. Tissue engineering based on the ability of ADSCs to differentiate into chondrocytes provides a new idea for the repair and regeneration of articular cartilage defects. The method for inducing the differentiation of ADSCs into chondrocytes in vitro who have been quite well established, which mainly include the use of growth factors and scaffolds to mimic the in vivo microenvironment, thereby promoting the differentiation of ADSCs into chondrocytes.


Assuntos
Cartilagem Articular , Células-Tronco , Adipócitos , Tecido Adiposo , Diferenciação Celular , Células Cultivadas , Condrócitos , Condrogênese , Engenharia Tecidual
4.
J Cell Physiol ; 234(7): 12029-12041, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30548623

RESUMO

Coronary atherosclerosis is a long-term, sustained, and evolving inflammatory disease manifested with the remodeling of the coronary arteries. The purpose of this study is to explore the potential role of microRNA-107 (miR-107) in vascular endothelial cells (VECs) in coronary atherosclerosis by regulating the KRT1 gene and the Notch signaling pathway. A mouse model of coronary atherosclerosis was established. The relationship between miR-107 and KRT1 was analyzed and verified by dual-luciferase reporter assay. The functional role of miR-107 in coronary atherosclerosis was determined using ectopic expression and depletion. Blood lipid levels and atherosclerotic index (AI) were measured in atherosclerotic mice. Expression pattern of miR-107, KRT1, Notch signaling pathway, inflammatory/anti-inflammatory factors, and endoplasmic reticulum (ER) stress-related genes was evaluated by means of reverse transcription quantitative polymerase chain reaction, western blot analysis, and enzyme-linked immunosorbent assay. Meanwhile, cell-cycle distribution and cell apoptosis in VECs were assessed by flow cytometry. Atherosclerotic mice exhibited higher blood lipid levels, AI, apoptotic index, and KRT1-positive expression as well as inhibited Notch signaling pathway when compared with normal mice. The miR-107 was revealed to bind to KRT1; miR-107 upregulation or KRT1 silencing resulted in reductions in blood lipid levels and AI, inhibition in cell apoptosis, inflammation, and ER stress. Restored miR-107 or downregulated KRT1 activated the Notch signaling pathway. These results supported the notion that miR-107-targeted KRT1 inhibition activated the Notch pathway, thereby, protecting against the coronary atherosclerosis. Findings in this study might provide a novel biomarker for the coronary atherosclerosis treatment.

5.
Cell Immunol ; 331: 30-37, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29773224

RESUMO

Early immunological events in acute HIV infection are thought to fundamentally influence long-term disease outcomes. Though the contribution of Gag-specific CD8 T cell responses to early viral control is well established, little is known about the role of Env-specific CD8 T cell responses in controlling viral replication during acute infection. In a macaque simian-human immunodeficiency virus (SHIV) model, some macaques who were able to control SHIV replication after ART interruption showed expansion of Env-specific CD8 T cell responses during acute infection, compared to macaques who progressed to viral rebound. To better understand the function of early Env-specific CD8 T cells, we isolated, expanded and examined their ability to act as effectors in vitro. We observed that Env-specific CD8 T cell clones have the capacity to directly recognize and kill SHIV-infected CD4 T cells, but failed to reduce viral replication in SHIV-infected macrophages. Our data suggest that early Env-specific CD8 T cell responses during acute SHIV infection contribute substantially to the control of viral replication. The T-cell clones composing of Env-specific effector cells demonstrates in vitro phenotypic and functional characteristics with the potentials to provide longlasting clinical benefit of in vivo HIV study.


Assuntos
Produtos do Gene env/imunologia , Macaca mulatta/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T Citotóxicos/imunologia , Replicação Viral/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/virologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/virologia , Humanos , Macaca mulatta/virologia , Macrófagos/imunologia , Macrófagos/virologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/fisiologia , Linfócitos T Citotóxicos/virologia , Carga Viral/imunologia
6.
Theriogenology ; 105: 15-26, 2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-28923703

RESUMO

Ovary development is a complex process involving numerous genes; the molecular mechanism underlying the ovary development of carp is still unknown. Here we used Illumina HiSeq™ 2500 to explore the transcriptome of undifferentiated gland (PG), juvenile ovary (OJ) and adult ovary (OA) of Yellow River carp (Cyprinus carpio). A total of 58,749 unigenes were obtained, comprising 45,707 known genes and 13,042 new genes. We identified differentially-expressed genes (DEGs) during development and characterized the functional properties of DEGs by comparison with the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes databases. qRT-PCR was used to analyze the expression of 22 DEGs and the results corresponded with those of RNA-Seq. Among DEGs between PG and OJ, some upstream regulators of gonad development were upregulated in PG, such as cyp19a and sox9, while some oocyte-specific genes were upregulated in OJ, such as nobox, bmp15 and zp2. Among DEGs between OJ and OA, many oocyte physiological function-related genes were upregulated in OA, such as fem-1 and foxl2. GO analysis showed a higher number of DEGs from PG-OJ analysis were assigned to reproduction terms. Furthermore, our investigation has also revealed DEGs identified from PG-OJ analysis were enriched in several important functional pathways, such as Fanconi anemia and the notch signal pathway. These data suggested a dynamic shift in gene expression during ovary development, and DEGs between PG and OJ provided crucial candidate gene data for the study of ovarian differentiation. Additionally, a total of 1,776,769 single nucleotide polymorphisms and 157,279 INDEs were revealed from transcriptome data. This result will contribute to knowledge of ovary differentiation of Yellow River carp.


Assuntos
Carpas/fisiologia , Perfilação da Expressão Gênica/veterinária , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Ovário/fisiologia , Maturidade Sexual/fisiologia , Transcriptoma/fisiologia , Processamento Alternativo , Animais , Feminino , Biblioteca Gênica , Mutação
7.
Fish Physiol Biochem ; 44(1): 375-386, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29164452

RESUMO

The suh gene is crucial in Notch pathway and regulates mammalian gonad development. In this study, the sequences of suh1 and suh2 genes in Yellow River carp (Cyprinus carpio) were verified. The partial 5'-flanking regions of suh1 and suh2 were analyzed and several potential transcription factor-binding sites were identified. Phylogenetic, gene structure, and chromosome synteny analyses revealed that carp suh1 and suh2 were orthologs and homologous to vertebrate suh. Investigation of the expression profiles of suh1 and suh2 with qPCR showed that these genes were abundant in the brain and gonad of carp, with suh1 exhibiting sexual dimorphism expression pattern in gonad. To study the relationship between gonad differentiation and Notch signaling, primordial gonads were exposed to DAPT, an inhibitor of Notch signaling, in vitro and in vivo. The results revealed a significant downregulation of suh1 and other Notch genes in vitro. In addition, expression of male-biased genes, such as amh, dmrt1, etc., was downregulated, whereas that of female-biased genes, such as foxl2, gdf9, etc., was upregulated. When the primordial gonads were subjected to long-term DAPT exposure, an increased proportion of ovary and delay in testis development were observed. These results suggest that suh gene may have a conservative function between teleosts and mammals. Furthermore, Notch signaling was found to be involved in gonad differentiation in Yellow River carp, and DAPT was noted to inhibit and enhance the expression of male- and female-biased genes, respectively, and induce the increase in number of females.


Assuntos
Carpas/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Gônadas/crescimento & desenvolvimento , Receptores Notch/metabolismo , Transdução de Sinais/fisiologia , Animais , Mapeamento Cromossômico , Cromossomos/genética , Feminino , Genômica , Masculino , Filogenia , Receptores Notch/genética , Sintenia
8.
Oncotarget ; 8(34): 55901-55914, 2017 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-28915561

RESUMO

Postoperative cognitive dysfunction (POCD) is a common postoperative complication observed in elderly patients. Using microarray analyses, we comprehensively compared long non-coding RNA (lncRNA), messenger RNA (mRNA), and microRNA (miRNA) expression profiles in hippocampal tissues from a mouse model of POCD and control mice. A total of 175 lncRNAs, 117 mRNAs, and 26 miRNAs were differentially expressed between POCD and control mice. Gene ontology (GO) and KEGG pathway enrichment analyses were performed to explore the principal functions of dysregulated genes. Correlated coding-noncoding co-expression (CNC) and competing endogenous RNA (ceRNA) expression networks were constructed using bioinformatics methods. lncRNA NONMMUT000708 correlated positively with expression of the inflammation-related gene Hif3a. lncRNAs NONMMUT043249 and NONMMUT028705 mediated gene expression by binding the transcription factor cAMP response element-binding protein (CREB). The constructed ceRNA network suggested lncRNA NONMMUT055714 binds competitively with miR-7684-5p, increasing expression of its target gene, Sorl1. Finally, eight dysregulated lncRNAs, four miRNAs, and ten mRNAs were confirmed via quantitative real-time polymerase chain reaction (PCR) in 10 POCD-healthy mouse paired samples. These results suggest that lncRNAs and miRNAs are involved in POCD pathogenesis and progression. Our ceRNA network will improve understanding of lncRNA-mediated ceRNA regulatory mechanisms operating during the pathogenesis of POCD.

9.
Injury ; 48(10): 2354-2359, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28847589

RESUMO

The treatment of subtrochanteric fractures is a challenge for orthopaedic trauma surgeons. Three positions have been described previously: supine on a fracture table, supine on a flat radiolucent table, and the lateral decubitus position on a flat radiolucent table. Each one has its advantages and limitations. In this article we describe a prone position for intramedullary nailing of subtrochanteric femoral fractures. This position has the advantages including: 1) an easy approach to reduce and maintain the reduction of fracture by adjusting only the leg plate on injured side, 2) perfect intraoperation fluoroscopic imaging on both anteroposterior view and lateral view, and 3) an easy approach to establish an appropriate entry point even in obese patients.


Assuntos
Pinos Ortopédicos , Fluoroscopia , Fixação Intramedular de Fraturas/métodos , Fraturas do Quadril/cirurgia , Decúbito Ventral , Idoso de 80 Anos ou mais , Placas Ósseas , Feminino , Fixação Intramedular de Fraturas/instrumentação , Fraturas do Quadril/diagnóstico por imagem , Fraturas do Quadril/fisiopatologia , Humanos , Resultado do Tratamento
10.
Virus Res ; 233: 8-16, 2017 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-28279801

RESUMO

Fitness is a key parameter in the measurement of transmission capacity of individual drug-resistant HIV. Drug-resistance related mutations (DRMs) T369V/I and A371V in the connection subdomain (CN) of reverse transcriptase (RT) occur at higher frequencies in the individuals experiencing antiretroviral therapy failure. Here, we evaluated the effects of T369V/I and A371V on viral fitness, in the presence or in the absence of thymidine analogue resistance-associated mutations (TAMs) and assessed the effect of potential RT structure-related mechanism on change in viral fitness. Mutations T369V/I, A371V, alone or in combination with TAMs were introduced into a modified HIV-1 infectious clone AT1 by site-directed mutagenesis. Then, experiments on mutant and wild-type virus AT2 were performed separately using a growth-competition assay, and then the relative fitness was calculated. Structural analysis of RT was conducted using Pymol software. Results showed that T369V/I severely impaired the relative virus fitness, and A371V compensated for the viral fitness reduction caused by TAMs. Structural modeling of RT suggests that T369V/I substitutions disrupt powerful hydrogen bonds formed by T369 and V365 in p51 and p66. This study indicates that the secondary DRMs within CN might efficiently damage viral fitness, and provides valuable information for clinical surveillance and prevention of HIV-1 strains carrying these DRMs.


Assuntos
Farmacorresistência Viral/genética , Aptidão Genética , Transcriptase Reversa do HIV/genética , Mutação , Nevirapina/química , Inibidores da Transcriptase Reversa/química , Substituição de Aminoácidos , Linhagem Celular , Expressão Gênica , Células HEK293 , Transcriptase Reversa do HIV/química , Transcriptase Reversa do HIV/metabolismo , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , HIV-1/genética , Humanos , Ligações de Hidrogênio , Modelos Moleculares , Mutagênese Sítio-Dirigida , Nevirapina/farmacologia , Domínios Proteicos , Inibidores da Transcriptase Reversa/farmacologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/virologia
11.
Immunity ; 44(4): 939-50, 2016 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-27067056

RESUMO

VRC01-class antibodies neutralize diverse HIV-1 strains by targeting the conserved CD4-binding site. Despite extensive investigations, crucial events in the early stage of VRC01 development remain elusive. We demonstrated how VRC01-class antibodies emerged in a Chinese donor by antigen-specific single B cell sorting, structural and functional studies, and longitudinal antibody and virus repertoire analyses. A monoclonal antibody DRVIA7 with modest neutralizing breadth was isolated that displayed a subset of VRC01 signatures. X-ray and EM structures revealed a VRC01-like angle of approach, but less favorable interactions between the DRVIA7 light-chain CDR1 and the N terminus with N276 and V5 glycans of gp120. Although the DRVIA7 lineage was unable to acquire broad neutralization, longitudinal analysis revealed a repertoire-encoded VRC01 light-chain CDR3 signature and VRC01-like neutralizing heavy-chain precursors that rapidly matured within 2 years. Thus, light chain accommodation of the glycan shield should be taken into account in vaccine design targeting this conserved site of vulnerability.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/imunologia , Sítios de Ligação de Anticorpos/imunologia , Anticorpos Anti-HIV/imunologia , Proteína gp120 do Envelope de HIV/imunologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia , Sequência de Aminoácidos , Linfócitos T CD4-Positivos/imunologia , Infecções por HIV/imunologia , HIV-1/imunologia , Humanos , Dados de Sequência Molecular
12.
Cell Immunol ; 303: 55-65, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27062692

RESUMO

Interleukin-21 (IL-21), which belongs to IL-2 γ chain receptor cytokine family, is as an important regulator of immune responses. In this study, we developed a novel strategy for immunizing mice with a DNA/vaccinia/protein vaccine in the presence or absence of mouse IL-21 (mIL-21) to evaluate whether mIL-21 could enhance immune responses. Our results demonstrated that co-immunization with mIL-21 did not increase significantly the capacity of vaccine induced antibodies to bind to HIV-1 GP140. An effect of mIL-21 in adjusting the efficacy of HIV-1 vaccine through enhancing Th1 type immune response was however observed. The frequencies of HIV-1-specific cytokine-producing CD4+ T and CD4+ TEM cells, especially multifunctional T cell responses, were significantly increased by co-administrating with mIL-21. A significant increase was also observed in the frequency of NK cells in mIL-21 adjuvant groups. Taken together, combination of mIL-21 with HIV-1 vaccines led to distinct enhancement of NK cells and T cell immune responses associated with immune protection.


Assuntos
Vacinas contra a AIDS/imunologia , Adjuvantes Imunológicos/uso terapêutico , Infecções por HIV/prevenção & controle , HIV-1/imunologia , Interleucinas/uso terapêutico , Células Matadoras Naturais/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Animais , Células Cultivadas , Feminino , Infecções por HIV/imunologia , Humanos , Células Matadoras Naturais/imunologia , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Linfócitos T/imunologia , Vacinas de DNA
13.
FEBS Lett ; 589(24 Pt B): 3938-44, 2015 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-26592151

RESUMO

It has been reported that cystatin c (Cys C) closely correlates with metabolic disorders such as obesity and diabetes. However, it is still unknown whether Cys C plays a role for these disorders. Our results showed that the insulin signal transduction was largely impaired by Cys C in hepatocytes. In myotubes, however, the insulin signal transduction was not affected. Following experiments revealed that Cys C could induce endoplasmic reticulum stress (ER stress) in hepatocytes, whereas Cys C had no such an effect in myotubes. The alleviation of ER stress by 4-Phenyl butyric acid (4-PBA) restored the impaired insulin signal transduction in Cys C-treated hepatocytes. These results provided direct evidence that, by inducing ER stress, Cys C impairs insulin signal transduction in hepatocytes.


Assuntos
Cistatina C/metabolismo , Regulação para Baixo , Estresse do Retículo Endoplasmático , Hepatócitos/metabolismo , Insulina/metabolismo , Transdução de Sinais , Animais , Antineoplásicos/farmacologia , Linhagem Celular , Células Cultivadas , Cistatina C/antagonistas & inibidores , Cistatina C/sangue , Cistatina C/genética , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/metabolismo , Regulação para Baixo/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/patologia , Humanos , Insulina/química , Insulina/genética , Masculino , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Fibras Musculares Esqueléticas/efeitos dos fármacos , Fibras Musculares Esqueléticas/metabolismo , Especificidade de Órgãos , Fenilbutiratos/farmacologia , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos
14.
PLoS One ; 9(12): e115422, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25522008

RESUMO

Avian leukosis virus subgroup J (ALV-J) has induced serious clinical outbreaks and has become a serious infectious disease of chickens in China. We describe here the creation of a recombinant ALV-J tagged with the enhanced green fluorescent protein (named rHPRS-103EGFP). We successfully utilize the rHPRS-103EGFP to visualize viral infection and for development of a simplified serum-neutralization test.


Assuntos
Anticorpos Neutralizantes/genética , Vírus da Leucose Aviária/imunologia , Testes de Neutralização/métodos , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/genética , Anticorpos Antivirais/imunologia , Leucose Aviária/imunologia , Leucose Aviária/virologia , Vírus da Leucose Aviária/genética , Linhagem Celular Tumoral , Galinhas
15.
Oncol Lett ; 7(5): 1645-1650, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24765193

RESUMO

Neddylation promotes the process of ubiquitination, which plays a critical role in the degradation of numerous proteins, including cell cycle and apoptosis regulators. In our previous study, an increase in neddylation was identified in melanoma cell lines. In the present study, the upregulation of neddylation was detected in melanoma tissues which confirmed the results of our previous study on melanoma cell lines. To explore the mechanism by which the process of neddylation was increased, the enzymes that regulate the process were investigated. These neddylation-related regulatory enzymes are potential targets for melanoma therapy. Downregulation of UBA3, a subunit of the E1 enzyme, by RNA interference caused cell cycle arrest at G0/G1 in the M14 cell line. In addition, cyclin D expression declined, whereas p27, p21 and bax expression increased. These findings suggest that interfering with the neddylation pathway may decrease the proliferation of melanoma through the modulation of cell cycle regulators and apoptosis promoters.

16.
PLoS One ; 9(1): e84797, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24465434

RESUMO

Subgroup J avian leukosis virus (ALV-J) was first isolated from meat-type chickens that had developed myeloid leukosis and since 2008, ALV-J infections in chickens have become widespread in China. A comparison of the sequence of ALV-J epidemic isolates with HPRS-103, the ALV-J prototype virus, revealed several distinct features, one of which is a 19-nucleotide (nt) insertion in the leader sequence. To determine the role of the 19-nt insertion in ALV-J pathogenicity, a pair of viruses were constructed and rescued. The first virus was an ALV-J Chinese isolate (designated rSD1009) containing the 19-nt insertion in its leader sequence. The second virus was a clone, in which the leader sequence had a deleted 19-nt sequence (designated rSD1009△19). Compared with rSD1009△19, rSD1009 displayed a moderate growth advantage in vitro. However, no differences were demonstrated in either viral replication or oncogenicity between the two rescued viruses in chickens. These results indicated that the 19-nt insertion contributed to ALV-J replication in vitro but was not related to its pathogenicity in vivo.


Assuntos
Vírus da Leucose Aviária/metabolismo , Vírus da Leucose Aviária/patogenicidade , Proteínas Virais/fisiologia , Animais , Leucose Aviária/virologia , Vírus da Leucose Aviária/genética , Linhagem Celular , Galinhas , Doenças das Aves Domésticas/virologia , Proteínas Virais/genética , Virulência/genética , Virulência/fisiologia , Replicação Viral/genética , Replicação Viral/fisiologia
17.
J Gen Virol ; 94(Pt 10): 2287-96, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23907393

RESUMO

MicroRNAs (miRNAs) are a class of small regulatory non-coding RNAs that modulate gene expression at the post-transcriptional level, playing a crucial role in cell differentiation and development. Recently, some reports have demonstrated that a number of cellular miRNAs play a role during viral infection. In this study, a luciferase-reporter system carrying the 5' untranslated region (5' UTR) and 3' UTR of avian leukosis virus subgroup J (ALV-J) was used to determine whether cellular miRNAs are involved in ALV-J infection. The miRNA gga-miR-1650 was screened for its potential interaction with the 5' UTR of ALV-J and the ability to suppress luciferase-reporter activity. A mutational analysis of predicted gga-miR-1650-binding sites showed that the 5' and 3' ends of gga-miR-1650 contributed to the interaction between gga-miR-1650 and its target located at the 5' UTR. Overexpression of miRNA gga-miR-1650 was shown to downregulate the expression of the Gag protein and influence the replication of ALV-J through binding to the 5' UTR. Overall, this report provides the basis for the development of new strategies for anti-ALV-J intervention.


Assuntos
Vírus da Leucose Aviária/fisiologia , MicroRNAs/metabolismo , RNA Viral/metabolismo , Replicação Viral/fisiologia , Regiões 5' não Traduzidas/fisiologia , Animais , Vírus da Leucose Aviária/classificação , Vírus da Leucose Aviária/genética , Linhagem Celular , Regulação Viral da Expressão Gênica , Humanos , MicroRNAs/genética , Mutação , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
18.
J Microbiol Biotechnol ; 23(5): 630-6, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23648851

RESUMO

Recently, there has been a growing body of evidence showing that cellular microRNAs (miRNAs) are involved in virus-host interactions. Numerous studies have focused on analyses of the expression profiles of cellular miRNAs, but the expression patterns of Dicer, which is responsible for the generation of miRNAs, have only rarely been explored in Gallus gallus. We developed a duplex realtime reverse transcriptase polymerase chain reaction (RTPCR) assay for the relative quantification of the mRNAs of Dicer and beta-actin in G. gallus. To apply this method, the expression of Dicer in avian cells after infection with avian leukosis virus subgroup J (ALV-J) was detected using our established duplex real-time RT-PCR. The duplex realtime RT-PCR assay is sufficiently sensitive, specific, accurate, reproducible, and cost-effective for the detection of Dicer in G. gallus. Furthermore, this study, for the first time, demonstrated that ALV-J can induce differential expression of Dicer mRNA in the ALV-J-infected cells.


Assuntos
Galinhas/genética , Doenças das Aves Domésticas/virologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Ribonuclease III/genética , Animais , Leucose Aviária/virologia , Vírus da Leucose Aviária/fisiologia , Galinhas/metabolismo , Galinhas/virologia , Ribonuclease III/metabolismo
19.
Vet Microbiol ; 162(1): 232-8, 2013 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-23157947

RESUMO

Avian leukosis virus subgroup J (ALV-J) has become pandemic and induced serious clinical outbreaks in chickens in China. In particular, ALV-J induced various clinical tumors in infected chickens, which caused enormous economic losses to poultry. In this study, an infectious clone from an epidemic ALV-J Chinese isolate designated HLJ09SH01 was constructed and rescued. The rescued virus (named rHLJ09SH01) was inoculated into specific-pathogen-free (SPF) layer chickens, and infected chickens were observed for 238 days to explore the oncogenicity of rHLJ09SH01. As a result, 57.9% of rHLJ09SH01-infected chickens produced tumors. Accumulating evidence shows that microRNAs (miRNAs) have a close relationship with tumorigenesis. To gain more insight into the tumorigenesis of ALV-J, a miRNA microarray was performed as part of an investigation of changes in host miRNA expression in a liver tumor from ALV-J infected chickens. The results showed that four miRNAs were significantly differentially expressed; these data were verified using real-time PCR. Bioinformatics analysis showed the differentially expressed miRNAs to be involved in some tumorigenesis-related signaling pathways, such as the MAPK signaling pathway and the Wnt signaling pathway, which may represent a possible signaling pathway that was involved in the ALV-J-induced tumorigenesis.


Assuntos
Vírus da Leucose Aviária/genética , Leucose Aviária/virologia , MicroRNAs/biossíntese , Doenças das Aves Domésticas/virologia , Animais , Leucose Aviária/patologia , Vírus da Leucose Aviária/isolamento & purificação , Vírus da Leucose Aviária/metabolismo , Carcinogênese , Embrião de Galinha , Galinhas , China , Genoma Viral , Carne/virologia , Redes e Vias Metabólicas/genética , MicroRNAs/genética , Análise de Sequência com Séries de Oligonucleotídeos , Provírus/genética , Organismos Livres de Patógenos Específicos
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