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Serine/threonine kinase receptor-associated protein (STRAP) serves as a scaffold protein and is engaged in a variety of cellular activities, although its importance in antiviral innate immunity is unknown. We discovered that STRAP works as an interferon (IFN)-inducible positive regulator, facilitating type I IFN signaling during pseudorabies virus infection. Mechanistically, STRAP interacts with TBK1 to activate type I IFN signaling. Both the CT and WD40 7 - 6 domains contribute to the function of STRAP. Furthermore, TBK1 competes with PRV-UL50 for binding to STRAP, and STRAP impedes the degradation of TBK1 mediated by PRV-UL50, thereby increasing the interaction between STRAP and TBK1. Overall, these findings reveal a previously unrecognized role for STRAP in innate antiviral immune responses during PRV infection. STRAP could be a potential therapeutic target for viral infectious diseases.
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Herpesvirus Suídeo 1 , Imunidade Inata , Interferon Tipo I , Proteínas Serina-Treonina Quinases , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Humanos , Herpesvirus Suídeo 1/imunologia , Interferon Tipo I/imunologia , Interferon Tipo I/metabolismo , Células HEK293 , Pseudorraiva/imunologia , Pseudorraiva/virologia , Transdução de Sinais , Linhagem Celular , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/imunologia , Proteínas de Ligação a RNA/metabolismo , Regulação para Cima , Ligação ProteicaRESUMO
The relationship between Parkinson's disease (PD) and urate or gout has attracted significant interest in recent years, but the results were conflicting. This dose-response meta-analysis aimed to estimate the correlation between urate levels or gout and the risk for PD. The Embase, PubMed, and Medline databases were searched for studies that investigated the relationship between the risk for PD and urate levels or gout. Random-effects or fixed-effects models were used to obtain pooled relative risks (RRs) and corresponding 95% confidence intervals (CIs). Fifteen studies, involving 449,816 participants and 14,687 cases in total, were included in the meta-analysis. High serum urate levels were associated with decreased risk for PD (RR 0.44 [95% CI 0.32-0.55]). Subgroup analysis according to sex revealed a neuroprotective effect of high urate levels against PD among females (0.68 [95% CI 0.43-0.93]) and males (0.49 [95% CI 0.34-0.64]). The risk for PD was lowered by 6% (0.94 [95% CI 0.90-0.98]) for each 1 mg/dl increase in serum urate level and reduced by 13% (0.87 [95% CI 0.80-0.95]) with each 2 mg/dl increase in serum urate level. However, gout was not closely correlated with the risk for PD (0.97 [95% CI 0.85-1.09]). Higher serum urate levels reduced the risk for PD, which was decreased by 6% (relative risk reduction) for each 1 mg/dl increase in serum urate levels. And the results indicated that urate may exert protective effects against the development of PD.
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Interferon-stimulated gene 15 (ISG15) is strongly upregulated during viral infections and exerts pro-viral or antiviral actions. While many viruses combat host antiviral defenses by limiting ISG expression, PRV infection notably increases expression of ISG15. However, studies on the viral strategies to regulate ISG15-mediated antiviral responses are limited. Here, we demonstrate that PRV-induced free ISG15 and conjugated proteins accumulation require viral gene expression. Conjugation inhibition assays showed that ISG15 imposes its antiviral effects via unconjugated (free) ISG15 and restricts the viral release. Knockout of ISG15 in PK15 cells interferes with IFN-ß production by blocking IRF3 activation and promotes PRV replication. Mechanistically, ISG15 facilitates IFNα-mediated antiviral activity against PRV by accelerating the activation and nuclear translocation of STAT1 and STAT2. Furthermore, ISG15 facilitated STAT1/STAT2/IRF9 (ISGF3) formation and ISGF3-induced IFN-stimulated response elements (ISRE) activity for efficient gene transcription by directly interacting with STAT2. Significantly, ISG15 knockout mice displayed enhanced susceptibility to PRV, as evidenced by increased mortality and viral loads, as well as more severe pathology caused by excessive production of the inflammatory cytokines. Our studies establish the importance of free ISG15 in IFNα-induced antiviral immunity and in the control of viral infections.
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Herpesvirus Suídeo 1 , Viroses , Camundongos , Animais , Camundongos Knockout , Antivirais/farmacologia , Interferon-alfa/farmacologiaRESUMO
Type I interferon (IFN) plays an important role in the host defense against viral infection by inducing expression of interferon-stimulated genes (ISGs). In a previous study, we found that porcine interferon-stimulated gene 15 (ISG15) exhibited antiviral activity against PRV in vitro. To further investigate the antiviral function of ISG15 in vivo, we utilized ISG15 knockout (ISG15-/-) mice in this study. Here, we demonstrate that ISG15-/- mice were highly susceptible to PRV infection in vivo, as evidenced by a considerably reduced survival rate, enhanced viral replication and severe pathological lesions. However, we observed no significant difference between female and male infected WT and ISG15-/- mice. Moreover, ISG15-/- mice displayed attenuated antiviral protection as a result of considerably reduced expression of IFNß and relevant ISGs during PRV replication. Furthermore, excessive production of proinflammatory cytokines may be closely related to encephalitis and pneumonia. In further studies, we found that the enhanced sensitivity to PRV infection in ISG15-/- mice might be caused by reduced phosphorylation of STAT1 and STAT2, thereby inhibiting type I IFN-mediated antiviral activity. Based on these findings, we conclude that ISG15 is essential for host type I IFN-mediated antiviral response.
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Antivirais , Interferon Tipo I , Animais , Antivirais/metabolismo , Antivirais/farmacologia , Citocinas/metabolismo , Feminino , Interferon Tipo I/metabolismo , Interferon-alfa , Masculino , Camundongos , Camundongos Knockout , Suínos , Ubiquitinas/genética , Ubiquitinas/metabolismoRESUMO
A magnesium phosphate cement-based engineered cementitious composite (MPC-ECC) was developed using polyvinyl alcohol (PVA) fibers and fly ash. In this study, the bond behavior of MPC-ECC with ordinary concrete was evaluated through single and double shear bond strength tests. The effects of the water to solid mass ratio (W/S), the sand to binder mass ratio (S/B), the molar ratio of MgO to KH2PO4 (M/P), the fly ash content (F), the borax dosage (B), the volume fraction of PVA fibers (Vf), and curing age on the bond behavior of MPC-ECC with ordinary concrete were examined. The results showed that as the W/S increased, the single and double shear bond strengths were gradually reduced. As the S/B increased, the double shear bond strength increased; the single shear bond strength first decreased up to an S/B of 0.1 and then increased. With the increase of M/P, the single and double shear bond strengths increased. With the increase of F, the single shear bond strength first increased up to an F of 30% and then decreased; the double shear bond strength decreased. With the increase of B, the single and double shear bond strengths increased first and then decreased, and their strength reached its maximum at a B of 6%. The increase of Vf improved the single and double shear bond strengths. The research results can provide some technical guidance for repairing concrete structures with MPC-ECC.
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INTRODUCTION: N6-Methyladenosine (m6A), the most common and reversible mRNA modification, has attracted considerable attention recently, and accumulating evidence indicates it has an important role in the progression of ischemic stroke (IS). AREAS COVERED: We first reviewed m6A methylation modification enzymes, including m6A methyltransferases (METTL3, METTL14, and WTAP), demethylases (FTO and ALKBH5), m6A-binding proteins (YTH domain containing 1/2 [YTHDC1/2], YTHDF1/2/3, and insulin like growth factor 2 mRNA binding protein 1/2/3 [IGF2BP1/2/3]), and their-related functions. An alteration in the m6A methylation profile of IS has been reported and m6A is differentially expressed in IS. Thus, we then focused on the underlying mechanism of m6A methylation in IS and the involvement of atherosclerosis (AS), cerebral ischemia/reperfusion (IR) injury, inflammation, oxidative stress, and apoptosis. Furthermore, we also elucidated the effect of m6A-associated single-nucleotide polymorphisms (SNPs) on stroke and uncovered new causal variants for IS. The clinical application of m6A targeting drugs is still in its infancy and will be available in the future. EXPERT OPINION: Collectively, the information in the present review is a summary of the latest developments in m6A modification and highlights the mechanisms underlying IS pathogenesis, which may provide novel insights into the mechanisms and therapeutic targets for IS.
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AVC Isquêmico , Adenosina/análogos & derivados , Adenosina/química , Adenosina/genética , Adenosina/metabolismo , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Humanos , AVC Isquêmico/genética , Metilação , Metiltransferases/metabolismo , RNA Mensageiro/genéticaRESUMO
CircRNAs belong to a novel class of noncoding RNAs that are generated by exons of genes by alternative mRNA splicing and involved in pathophysiological processes of ischemic stroke by regulating neuro-inflammation. A total of 982 patients were enrolled in our study for stroke recovery analysis. The aim of our study was to first explore the association between the inflammation-related circRNA polymorphism and functional outcome 3 months after ischemic stroke by using multivariate logistic regression model. Next, we further investigated the role of circRNA polymorphism in predicting stroke recurrence by using Cox proportional hazard regression model. Five circRNA polymorphisms were genotyped by using polymerase chain reaction and ligation detection reaction method. We identified circ-STAT3 (signal transducer and activator of transcription) rs2293152 GG genotype to be associated with poorer recovery 90 days after stroke (OR = 1.452, 95% CI: 1.165-4.362, p = 0.016). After adjusting for confound factors, the association for rs2293152 with 3 months outcome after IS was stronger, suggesting a mechanism that rs2293152 is an independent risk factor for stroke recovery (OR = 2.255, 95% CI: 1.034-2.038, p = 0.031). However, no other circRNA polymorphisms (circ-DLGAP4 rs41274714, circ-TRAF2 rs10870141, circ-ITCH rs10485505, rs4911154) were associated with functional outcome 3 months after stroke in any genetic models. Subgroup analysis revealed that the negative effect of rs2293152 GG genotype was greater in female and older patients, subjects with history of hypertension. Additionally, all the circRNA polymorphisms were not correlated with recurrent risk of ischemic stroke. Our results indicated that circ-STAT3 might be a novel biomarker for predicting functional outcome after stroke and an important contributor to the ischemic stroke recovery.
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AVC Isquêmico/genética , Polimorfismo de Nucleotídeo Único , RNA Circular/genética , Biomarcadores/metabolismo , Feminino , Humanos , AVC Isquêmico/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteínas Repressoras/genética , Proteínas Associadas SAP90-PSD95/genética , Fator de Transcrição STAT3/genética , Fator 2 Associado a Receptor de TNF/genética , Ubiquitina-Proteína Ligases/genéticaRESUMO
Chemotherapy-induced peripheral neuropathy (CIPN) is a common side effect during the course of cancer treatment, which is mainly manifested as a series of sensory abnormalities. At present, there are no recommended prevention or treatment strategies, and the underlying mechanisms are unclear. The ketogenic diet (KD), a special diet that is high in fat and low in carbohydrate intake, shows good therapeutic potential in children with epilepsy. In this study, it was found that KD significantly prevented paclitaxel-induced neuropathic nociception. Using the GSE113941 database, 281 differentially expressed genes (DEGs) were found in an animal model of CIPN and controls. The DEGs were mainly enriched in peroxisome proliferator activated receptor (PPAR) and oxidative phosphorylation signalling pathways. As a main regulatory pathway of lipid metabolism, the PPARγ signalling pathway was significantly upregulated in the KD model. In addition, KD also inhibited the expression of pro-inflammatory cytokines and the TLR4/NF-κB signalling pathway in the dorsal root ganglion (DRG) in paclitaxel-treated rats. In vitro, rat primary DRG neurons were used to investigate the role of PPARγ in paclitaxel-induced neurotoxicity. It was found that PPARγ agonist rosiglitazone significantly protected DRG neurons against cell apoptosis and reactive oxygen species generation induced by paclitaxel administration. Therefore, KD is a prospective treatment option when applied as a dietary intervention in the prevention and treatment of paclitaxel-induced neuropathic nociception, possibly through the activation of PPARγ and its neuroprotective functions.
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Antineoplásicos Fitogênicos , Dieta Cetogênica , Doenças do Sistema Nervoso Periférico , Animais , Gânglios Espinais , Nociceptividade , PPAR gama , Paclitaxel/toxicidade , Estudos Prospectivos , Ratos , Ratos Sprague-DawleyRESUMO
Background: Recent studies have reported that homocysteine (Hcy) may play a vital role in the pathogenesis of vascular dementia (VaD) and Alzheimer's disease (AD). Our study explored the relationship between the plasma Hcy and folate levels and the risk of dementia. Methods: We searched Embase, PubMed, and Web of Science for published literature, including case-control studies and prospective cohort studies, and performed a systematic analysis. Results: The results of our meta-analysis, consisting of case-control studies, showed higher levels of Hcy and lower levels of folate in dementia, AD, and VaD patients than those in non-demented controls (for dementia: SMD = 0.812, 95% CI [0.689, 0.936], p = 0.000 for Hcy; SMD = -0.677, 95% CI [-0.828, -0.525], p = 0.000 for folate). AD patients showed significantly lower plasma Hcy levels compared to VaD patients (SMD = -0.278, 95% CI [-0.466, -0.09], p = 0.000). Subgroup analysis revealed that ethnicity, average age, and dementia type had no significant effect on this association. Furthermore, from the analysis of prospective cohort studies, we identified that elevated plasma Hcy levels were associated with an increased risk of dementia, AD, and VaD (RRdementia = 1.22, 95% CI [1.08, 1.36]; RRAD = 1.07, 95% CI [1.04, 1.11]; RRVaD = 1.13, 95% CI [1.04, 1.23]). In addition, every 5 µmol/L increase in the plasma Hcy level was associated with a 9% increased risk of dementia and a 12% increased risk of AD. Conclusion: Hcy and folic acid are potential predictors of the occurrence and development of AD. A better understanding of their function in dementia could provide evidence for clinicians to rationalize clinical intervention strategies.
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Getah virus (GETV), a mosquito-borne virus belonging to the Alphavirus genus of family Togaviridae, has become increasingly problematic, which poses a huge threat to the safety of animals and public health. In order to detect GETV quickly and accurately, we have developed a SYBR Green I real-time quantitative reverse transcription PCR (RT-qPCR) assay for GETV with the detection limit of 66 copies/µL, excellent correlation coefficient (R2) of 0.9975, and amplification efficiency (E) of 98.90%, the target selected was the non-structural protein 3 of GETV. The sensitivity of it was higher than that of ordinary RT-PCR by 1000 folds, and the inter-assay and intra-assay CV values were all less than 0.99%. The newly developed RT-qPCR assay exhibited good sensitivity and reproducibility, which will provide technical support for the reliable and specific rapid diagnosis, and quantitative analysis of GETV infection.
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Alphavirus , Culicidae , Alphavirus/genética , Animais , Benzotiazóis , Diaminas , Quinolinas , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Transcrição Reversa , Sensibilidade e Especificidade , SuínosRESUMO
Long noncoding RNAs (lncRNAs) serve as regulators or effectors of the p53 regulatory pathway. The lncRNA-p53 regulatory network plays an important role in ischemia-induced apoptosis and may be important for post-stroke recovery. Eight genetic variants of p53-related lncRNAs were genotyped in 982 patients to explore the association of single nucleotide polymorphisms (SNPs) in the genes related to the p53 regulatory pathway with ischemic stroke (IS) prognosis in a northern Chinese population. Long- and short-term outcomes were assessed by stroke recurrence and modified Rankin Scale score 3 months after stroke, respectively. We first identified that p53 rs1042522 and LINC-ROR rs2027701 could be associated with IS recurrence risk. On further cumulative effect analysis, we found that these two polymorphisms could jointly be associated with IS recurrence. Patients carrying 2-4 risk alleles showed a significantly higher IS recurrence risk than those harboring no or a single risk allele. In contrast to rs2027701 and rs1042522, the other SNPs were not associated with IS recurrence. Subsequently, we found that TUG1 rs2240183 CC genotype was associated with a favorable IS outcome after adjusting for confounding factors. However, the other seven genetic variants of p53-related lncRNAs were not associated with a functional outcome after stroke. p53 rs1042522 and LINC-ROR rs2027701 may exert combined effects on IS recurrence, and TUG1 rs2240183 may be a new biomarker to predict short-term IS outcomes as it modulates p53-mediated apoptosis.
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Isquemia Encefálica/genética , Estudos de Associação Genética/métodos , Variação Genética/genética , AVC Isquêmico/genética , RNA Longo não Codificante/genética , Proteína Supressora de Tumor p53/genética , Adulto , Idoso , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/epidemiologia , China/epidemiologia , Feminino , Humanos , AVC Isquêmico/diagnóstico , AVC Isquêmico/epidemiologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , PrognósticoRESUMO
BACKGROUND: Recent studies have reported that lncRNA (long noncoding RNAs) antisense non-coding RNA in the INK4 locus (ANRIL) plays important roles in the development of atherosclerosis through regulating cell apoptosis, proliferation, and adhesion. GWAS (genome-wide association studies) identified common genetic variants within ANRIL could confer risk of ischemic stroke (IS) in southern Sweden. METHODS: We performed a case-control study, including 567 IS patients and 552 healthy controls from unrelated northern Chinese Han population, aiming to explore the association between lncRNA ANRIL rs2383207, rs4977574 polymorphisms and the risk of IS. Subsequently we implemented a meta-analysis to further assess the relationship of these variants and the disease. RESULTS: In our case-control study, no significant associations were observed in all models between above 2 polymorphisms and IS. Next in our subgroup analysis, we detected significant association between GA genotype of rs4977574 and the increased risk of LAA-IS (large-artery atherosclerotic ischemic stroke), similar elevated risk also appeared in the GGâ+âGA genotype under the dominant model (Pâ=â.048, ORâ=â1.385, 95% CIs 1.002-1.914; Pâ=â.040, ORâ=â1.378, 95% CIs 1.015-1.872, respectively). As for rs2383207, negative results were obtained under all models and subgroups. Our meta-analysis showed a significant association between rs4977574 polymorphism and IS risk in allele model (G vs A Pâ=â.002, ORâ=â1.137, 95% CIs 1.048-1.234); with respect to rs2383207 polymorphism, no significant association between that and the risk of IS was detected under the dominant model (GAâ+âAA vs GG, Pâ=â.061, ORâ=â0.923, 95% CIs 0.849-1.004), or recessive model (AA vs GAâ+âGG, Pâ=â.656, ORâ=â0.972, 95% CIs 0.858-1.101), or allele model (A vs G, Pâ=â.326, ORâ=â0.952, 95% CIs 0.863-1.050). Likewise, no significant association between rs2383207 and IS was found in different stoke subtypes (Pâ>â.05). CONCLUSIONS: Our findings indicated G allele of lncRNA ANRIL rs4977574 could increase the risk of IS, and the variant may be associated with susceptibility to LAA-IS in Chinese Han population.
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Predisposição Genética para Doença/genética , AVC Isquêmico/genética , Polimorfismo Genético/genética , RNA Longo não Codificante/sangue , Idoso , Alelos , Povo Asiático/etnologia , Povo Asiático/genética , Aterosclerose/epidemiologia , Aterosclerose/etnologia , Aterosclerose/genética , Estudos de Casos e Controles , China/epidemiologia , China/etnologia , Doença da Artéria Coronariana/epidemiologia , Doença da Artéria Coronariana/etnologia , Doença da Artéria Coronariana/genética , Feminino , Predisposição Genética para Doença/etnologia , Estudo de Associação Genômica Ampla , Genótipo , Humanos , AVC Isquêmico/epidemiologia , AVC Isquêmico/etnologia , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
In September 2019, a highly prevalent infectious disease caused severe hydropericardium hepatitis syndrome (HHS) in a peacock farm in Central China. The disease showed high mortality of 78.6% in 28-42 day-old peacocks. In this study, one strain of highly pathogenic fowl adenovirus serotype 4 (FAdV-4) was isolated from peacocks and designated as HN19. Molecular characterization of amino acid revealed that HN19 contains the same deletions as the dominate strains in chickens in China recently. Phylogenetic analyses revealed that HN19 showed higher homology with other FAdV-4 strains isolated from China, indicating that HN19 might originate from previously FAdV-4 predecessor in China. Experimental infection of the HN19 strain via intramuscular injection led to 100% mortality rate in 21-day-old specific pathogenic-free (SPF) chickens. To our knowledge, this represents the first report on the prevalence of FAdV-4 in peacocks. These results suggested that the potential risk of cross-species transmission of FAdV-4 from chickens to peacocks, highlighting the need for implementing strict biosecurity measures to avoid the mixing of different bird species.
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Infecções por Adenoviridae/veterinária , Aviadenovirus/classificação , Aviadenovirus/patogenicidade , Galliformes , Derrame Pericárdico/veterinária , Doenças das Aves Domésticas/virologia , Infecções por Adenoviridae/virologia , Animais , Galinhas , Derrame Pericárdico/virologia , Organismos Livres de Patógenos Específicos , VirulênciaRESUMO
In the present study, Getah virus (GETV) isolate, GETV-V1, was isolated from a commercial PRRSV attenuated live vaccine (MLV), which has been widely used to immunize pigs against porcine reproductive and respiratory syndrome virus (PRRSV). Further analysis demonstrated that nine batches of the PRRSV MLV vaccine (three batches per year from 2017 to 2019) from the same manufacturer were all positive for GETV. Genomic analyses indicated that the GETV-V1 isolate shared the highest sequence identity with the GETV strain, 16-I-674, which was isolated from horses in Japan. The phylogenetic analysis based on the genomic sequences showed that the GETV-V1 strain was clustered with the Japanese GETV strains. Taken together, this is the first report of GETV contamination in live swine vaccines in China. Our findings demonstrate that immunization with commercial live vaccines might be a potential novel route of GETV transmission in swine. This highlights the need for more extensive monitoring of commercial live vaccines.
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Alphavirus/classificação , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vacinas Virais/análise , Alphavirus/genética , Alphavirus/isolamento & purificação , Animais , Linhagem Celular , China , Contaminação de Medicamentos , Cavalos , Japão , Filogenia , Filogeografia , SuínosRESUMO
Gallibacterium anatis (G. anatis), one of the major pathogens causing reproductive tract disorders in laying hens, leads to a reduction in egg production and increased mortality, caused by either single or mixed infections with other pathogens. As a specific virulence factor of G. anatis, the role of GtxA in layers' salpingitis remains unclear. In this study, we explored the effect of GtxA on G. anatis infection by comparing wild strain Yu-PDS-RZ-1-SLG (RZ) and its GtxA deleted counterpart RZΔgtxA in primary chicken oviduct epithelial cells (COEC). Their adherence, invasion, cytoxicity, and ability to induce apoptosis and and cytokine secretion were evaluated and the cytotoxicity and cytokine secretion of the recombinant GtxA protein and its N-terminal adenylate cyclase and C-terminal RTX hemolysin domain were also analyzed. We found that the adhesion ability of RZΔgtxA was significantly lower than that of parental strain RZ, and its toxicity to COEC was weakened; Meanwhile, apoptosis was inhibited and the expression of IL-6, IL-2, TNF-α and IFN-γ were dramatically reduced in COEC infected by RZΔgtxA. In contrast, the recombinant protein GtxA inhibited the proliferation of oviduct cells and induced obvious cytotoxicity, and the expression of IL-6, TNF-α and IFN-γ were up-regulated in COEC interacted with recombinant proteins. Our study indicates that GtxA promotes G. anatis adherence to cells, changes cells permeability and expression of inflammatory factors, resulting in cell damage and apoptosis.
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Toxinas Bacterianas/genética , Infecções por Pasteurellaceae/veterinária , Pasteurellaceae/patogenicidade , Doenças das Aves Domésticas/microbiologia , Animais , Aderência Bacteriana , Galinhas , Citocinas/metabolismo , Células Epiteliais/citologia , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Feminino , Deleção de Genes , Oviductos/citologia , Oviductos/imunologia , Oviductos/microbiologia , Pasteurellaceae/genética , Pasteurellaceae/imunologia , Infecções por Pasteurellaceae/imunologia , Fatores de Virulência/genéticaRESUMO
Viral-encoded microRNAs (miRNAs) have vital roles in the regulation of virus replications and host immune responses. The results of previous studies have indicated that miRNA clusters are involved in the replication and virulence of the pseudorabies virus (PRV), which may potentially lead to immune escape or facilitation of PRV replication. This study's previous research revealed that prv-miR-LLT11a was differentially expressed during PRV infection. The present study's results have demonstrated that prv-miR-LLT11a could significantly inhibit PRV replication. It was further determined that SLA-1 was the target gene of prv-miR-LLT11a, and simultaneously, that overexpression of prv-miR-LLT11a could downregulate the mRNA and protein levels of SLA-1 in a dose-independent manner. Furthermore, the present study also observed that prv-miR-LLT11a can downregulate TAP1 expression. Our findings provide a better understanding of the molecular mechanism involved in the effects of prv-miR-LLT11a on SLA-1 and TAP1 as well as its involvement in immune system evasion of PRV.
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Herpesvirus Suídeo 1/fisiologia , Antígenos de Histocompatibilidade Classe I/genética , MicroRNAs/genética , RNA Viral/genética , Replicação Viral/genética , Animais , Linhagem Celular , Regulação para Baixo , Herpesvirus Suídeo 1/genética , MicroRNAs/metabolismo , RNA Viral/metabolismo , Sus scrofa , Regulação para CimaRESUMO
Pseudorabies virus (PRV) is one of the most vital pathogens of swine, leading to huge economic losses to the pig industry. Functioning in innate immunity, type-I interferon (IFN) plays a vital role in initial stage of viral infection. ISG15, an IFN-stimulated ubiquitin-like protein, is highly increased during virus infection and participates in the IFN-mediated antiviral immune response. However, limited attention has been paid to the functional role of porcine ISG15 (pISG15) in PRV infection. In this study, we generated a PK15 inducible cell line stably expressing the pISG15 gene and investigated the potential anti-PRV response of pISG15. We demonstrated that pISG15 was upregulated in an early stage of PRV infection, and pISG15 overexpression efficiently inhibited PRV replication by reducing the viral titers and mRNA levels of PRV, and also increased expression of IFN-ß and activation of the ISRE promoter. However, knockdown of pISG15 by siRNA did not affect PRV replication, and potentiated IFN-I-mediated signaling, resulting in an increase in antiviral response in the process of PRV infection. The results showed that pISG15 has a potential immunodulatory role in cellular antiviral response against PRV.
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Citocinas/metabolismo , Herpesvirus Suídeo 1/fisiologia , Pseudorraiva/imunologia , Regulação para Cima , Animais , Linhagem Celular , Chlorocebus aethiops , Resistência à Doença , Herpesvirus Suídeo 1/imunologia , Interferon beta/metabolismo , Pseudorraiva/virologia , Suínos , Doenças dos Suínos/metabolismo , Doenças dos Suínos/virologia , Células Vero , Carga Viral , Replicação ViralRESUMO
In order to evaluate the genetic variability of encephalomyocarditis virus (EMCV), the whole genomes of six EMCV field isolates originating from pigs and rats origin in different regions of central China, were phylogenetically and comparatively analyzed. Phylogenetic analysis of whole genome sequences, open reading frame (ORF), capsid coding region (CCR) and VP3/VP1 using neighbor-joining analysis revealed that these isolates belonged to lineage 1. Nucleotide sequences of six isolates showed more than 99% pairwise identity rates, and the sequences of isolates from pig and boar in the same region were completely identical with each other, without any genetic deletion or insertion. From comparative analysis of variability of each EMCV protein coding region, 3D and VP3 regions showed that the highest average identity rates, and was confirmed as highly conserved. In contrast, the protein coding regions 3A and 3B was confirmed to be highly variable region with the lowest average identity rate. Our data confirmed that the EMCV strains isolated from pigs and rats origin had high homology with each other, which implied rats may play an important role in EMCV transmission between domestic pigs and wild boars.
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Infecções por Cardiovirus/veterinária , Vírus da Encefalomiocardite/classificação , Vírus da Encefalomiocardite/genética , Genoma Viral , Filogenia , Doenças dos Suínos/virologia , Sequenciamento Completo do Genoma , Animais , Genômica/métodos , Ratos , SuínosRESUMO
Porcine epidemic diarrhea virus (PEDV) inhibits the host typeâ interferon and cellular antiviral response, but its inhibition mechanism is unclear, and the roles of PEDV nonstructural proteins in regulating typeâ interferon responses have been seldom studied. To study the effect of nsp1 on typeâ interferon response, nsp1 gene was cloned into a eukaryotic expression vector pCAGGS. The expression of nsp1 in transfected cells was determined by Western blot and indirect immunofluorescence assay. The effects of nsp1 on the induction of typeâ interferon were evaluated by dual luciferase reporter gene assay, ELISA and VSV bioassay. Western blot and indirect immunofluorescence assay showed that nsp1 was highly expressed in transfected cells and PEDV-infected cells. Dual luciferase reporter gene assay results indicated that nsp1 strongly inhibited the IFN-ß promoter activity, and the inhibitory effect was nsp1 dose-dependent. ELISA results showed that nsp1 significantly inhibited the expression of IFN-ß in protein level. And VSV replication-inhibition bioassay revealed that nsp1 significantly inhibited typeâ IFN antiviral activities induced by poly(I:C). Our results implied that nsp1 was a highly conserved protein of PEDV and exhibited antagonistic function on interferon promoter activity. The results have laid a foundation for further understanding the immune evasion mechanism of PEDV and for developing new effective vaccine against PEDV.
Assuntos
Infecções por Coronavirus/veterinária , Interferon beta/imunologia , Doenças dos Suínos/imunologia , Suínos/imunologia , Proteínas não Estruturais Virais/imunologia , Animais , Infecções por Coronavirus/imunologia , Evasão da Resposta Imune , Vírus da Diarreia Epidêmica Suína , Regiões Promotoras Genéticas , Suínos/virologia , Doenças dos Suínos/virologiaRESUMO
Gallibacterium anatis (G. anatis) has been suggested to have a causal role in salpingitis and peritonitis in egg-laying chickens, leading to decreased egg production and increased mortality worldwide. Adherence and invasion of epithelial cells are thought to play a role in the pathogenesis of G. anatis infection. The purpose of this article was to study adherence and invasion of G. anatis using two G. anatis strains of different virulence (Yu-PDS-RZ-1-SLG strain, highly virulent and F149T strain, non-virulent) via infection of the primary chicken oviduct epithelial cells (PCOECs).The results showed that Yu-PDS-RZ-1 -SLG strain was able to attach to PCOECs at higher levels than that of F149T strain, but no invasion was observed with either strain. However, cell debris and cell apoptosis were observed after being exposed to G. anatis Yu-PDS-RZ-1-SLG for 90min, whereas G. anatis F149T did not cause cell damage, and adherence was prevented by trypsin treatment of bacterial cells. Cytokines were detected by ELISA after infection, and the results showed that the expression of IL-6, TNF-α, and IFN-γ levels was higher in virulent strain infection than that of the avirulent group. Results also indicated that the highly virulent strain G. anatis displayed an increased level of adherence. Changes in cytokine profiles in this study suggested that the production of cytokines might influence the microenvironment of oviduct and promote adherence, serving as a possible mechanism inducing cell damage.