RESUMO
Foot-and-mouth disease virus (FMDV) is a positive-strand RNA virus of the family Picornaviridae. Early studies show that some viruses of Picornaviridae, such as EMCV and EV71, induce NLRP3 inflammasome activation. Our current study demonstrates that FMDV induces the secretion of caspase-1 and interleukin 1 beta (IL-1ß), as well as activates the NLRP3 inflammasome in a dose- and time-dependent manner. Meanwhile, NLRP3 inflammasome can suppress FMDV replication during virus infection. Both FMDV RNA and viroporin 2B stimulate NLRP3 inflammasome activation. FMDV RNA triggers NLRP3 inflammasome through p-NF-κB/p65 pathway not dependent on RIG-I inflammasome. FMDV 2B activates NLRP3 inflammasome through elevation of intracellular ion, but not dependent on mitochondrial reactive oxygen species (ROS) and lysosomal cathepsin B. It further demonstrates that 2B viroporin activates NLRP3 inflammasome and induces IL-1ß in mice, which enhances the specific immune response against FMDV as an ideal self-adjuvant for FMD VLPs vaccine in guinea pigs. The results reveal a series of regulations between NLRP3 inflammasome complex and FMDV. Amino acids 140-145 of 2B is essential for forming an ion channel. By mutating the amino acid and changing the hydrophobic properties, the helical transmembrane region of the viroporin 2B is altered, so that the 2B is insufficient to trigger the activation of NLRP3 inflammasome. This study demonstrates the functions of FMDV RNA and 2B viroporin activate NLRP3 inflammasome and provides some useful information for the development of FMD vaccine self-adjuvant, which is also helpful for the establishment of effective prevention strategies by targeting NLRP3 inflammasome.
Assuntos
Vírus da Febre Aftosa/patogenicidade , Febre Aftosa/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteínas não Estruturais Virais/metabolismo , Animais , Feminino , Febre Aftosa/virologia , Vírus da Febre Aftosa/genética , Vírus da Febre Aftosa/metabolismo , Cobaias , Interações Hospedeiro-Patógeno/fisiologia , Inflamassomos/metabolismo , Interleucina-1beta/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Células RAW 264.7 , RNA Viral/metabolismo , Proteínas Viroporinas/química , Proteínas Viroporinas/metabolismoRESUMO
Peste des petits ruminants virus (PPRV), belonging to paramyxoviruses, has six structure proteins (such as matrix protein (M), nucleocapsid proteins (N), fusion protein (F) and hemagglutinin protein (H)) and could cause high morbidity and mortality in sheep and goats. Although a vaccine strain of PPRV has been rescued and co-expression of M and N could yield PPRV-like particles, the roles of structure proteins in virion assembly and release have not been investigated in detail. In this study, plasmids carrying PPRV cDNA sequences encoding the N, M, H, and F proteins were expressed in Vero cells. The co-expression of all four proteins resulted in the release of virus-like particles (VLPs) with similar release efficiency to that of authentic virions. Moreover, the co-expression of M together with F also resulted in efficient VLPs release. In the absence of M protein, the expression of no combination of the other proteins resulted in particle release. In summary, a VLPs production system for PPRV has been established and M protein is necessary for promoting the assembly and release of VLPs, of which the predominant protein is M protein. Further study will be focused on the immunogenicity of the VLPs.
Assuntos
Vírus da Peste dos Pequenos Ruminantes/metabolismo , Vírus da Peste dos Pequenos Ruminantes/fisiologia , Células Vero/metabolismo , Proteínas da Matriz Viral/metabolismo , Animais , Anticorpos Antivirais , Chlorocebus aethiops/metabolismo , Chlorocebus aethiops/fisiologia , DNA Complementar , DNA Viral , Hemaglutininas Virais/metabolismo , Hemaglutininas Virais/fisiologia , Camundongos , Proteínas do Nucleocapsídeo/metabolismo , Proteínas do Nucleocapsídeo/fisiologia , Vírus da Peste dos Pequenos Ruminantes/genética , Vírus da Peste dos Pequenos Ruminantes/imunologia , Proteínas Virais de Fusão/metabolismo , Proteínas Virais de Fusão/fisiologiaRESUMO
BACKGROUND: Peste des petits ruminants (PPR) is an economically important, Office International des Epizooties (OIE) notifiable, transboundary viral disease of small ruminants such as sheep and goat. PPR virus (PPRV), a negative-sense single-stranded RNA virus, is the causal agent of PPR. Therefore, sensitive, specific and rapid diagnostic assay for the detection of PPRV are necessary to accurately and promptly diagnose suspected case of PPR. METHODS: In this study, reverse transcription recombinase polymerase amplification assays using real-time fluorescent detection (real-time RT-RPA assay) and lateral flow strip detection (LFS RT-RPA assay) were developed targeting the N gene of PPRV. RESULTS: The sensitivity of the developed real-time RT-RPA assay was as low as 100 copies per reaction within 7 min at 40 °C with 95% reliability; while the sensitivity of the developed LFS RT-RPA assay was as low as 150 copies per reaction at 39 °C in less than 25 min. In both assays, there were no cross-reactions with sheep and goat pox viruses, foot-and-mouth disease virus and Orf virus. CONCLUSIONS: These features make RPA assay promising candidates either in field use or as a point of care diagnostic technique.
Assuntos
Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/veterinária , Peste dos Pequenos Ruminantes/diagnóstico , Vírus da Peste dos Pequenos Ruminantes/genética , Vírus da Peste dos Pequenos Ruminantes/isolamento & purificação , Animais , Reações Cruzadas , Corantes Fluorescentes , Vírus da Febre Aftosa/genética , Vírus da Febre Aftosa/isolamento & purificação , Doenças das Cabras/diagnóstico , Doenças das Cabras/virologia , Cabras/virologia , Vírus do Orf/genética , Vírus do Orf/isolamento & purificação , Peste dos Pequenos Ruminantes/virologia , RNA Viral/isolamento & purificação , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Reversa , Sensibilidade e Especificidade , Ovinos/virologia , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/virologiaRESUMO
BACKGROUND: Foot-and-mouth disease virus (FMDV) causes a severe vesicular disease in domestic and wild cloven-hoofed animals. Because of the limited early protection induced by current vaccines, emergency antiviral strategies to control the rapid spread of FMD outbreaks are needed.Here we constructed multiple microRNAs (miRNAs) targeting the internal ribosome entry site (IRES) element of FMDV and investigated the effect of IRES-specific miRNAs on FMDV replication in baby hamster kidney (BHK-21) cells and suckling mice. RESULTS: Four IRES-specific miRNAs significantly reduced enhanced green fluorescent protein (EGFP) expression from IRES-EGFP reporter plasmids, which were used with each miRNA expression plasmid in co-transfection of BHK-21 cells. Furthermore, treatment of BHK-21 cells with Bi-miRNA (a mixture of two miRNA expression plasmids) and Dual-miRNA (a co-cistronic expression plasmid containing two miRNA hairpin structures) induced more efficient and greater inhibition of EGFP expression than did plasmids carrying single miRNA sequences.Stably transformed BHK-21 cells and goat fibroblasts with an integrating IRES-specific Dual-miRNA were generated, and real-time quantitative RT-PCR showed that the Dual-miRNA was able to effectively inhibit the replication of FMDV (except for the Mya98 strain) in the stably transformed BHK-21 cells.The Dual-miRNA plasmid significantly delayed the deaths of suckling mice challenged with 50× and 100× the 50% lethal dose (LD50) of FMDV vaccine strains of three serotypes (O, A and Asia 1), and induced partial/complete protection against the prevalent PanAsia-1 and Mya98 strains of FMDV serotype O. CONCLUSION: These data demonstrate that IRES-specific miRNAs can significantly inhibit FMDV infection in vitro and in vivo.
Assuntos
Antivirais/metabolismo , Produtos Biológicos/metabolismo , Vírus da Febre Aftosa/fisiologia , MicroRNAs/metabolismo , Biossíntese de Proteínas , Ribossomos/metabolismo , Replicação Viral , Animais , Fusão Gênica Artificial , Sítios de Ligação , Células Cultivadas , Cricetinae , Modelos Animais de Doenças , Febre Aftosa/terapia , Vírus da Febre Aftosa/genética , Genes Reporter , Cabras , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Camundongos , MicroRNAs/genética , Reação em Cadeia da Polimerase em Tempo Real , Ribossomos/genética , Análise de Sobrevida , Resultado do TratamentoRESUMO
Wnt proteins are a family of secreted glycoproteins that are evolutionarily conserved and considered to be involved in extensive developmental processes in metazoan organisms. The characterization of wnt genes may improve understanding the parasite's development. In the present study, a wnt4 gene encoding 491amino acids was amplified from cDNA of metacestodes of Taenia solium using reverse transcription PCR (RT-PCR). Bioinformatics tools were used for sequence analysis. The conserved domain of the wnt gene family was predicted. The expression profile of Wnt4 was investigated using real-time PCR. Wnt4 expression was found to be dramatically increased in scolex evaginated cysticerci when compared to invaginated cysticerci. In situ hybridization showed that wnt4 gene was distributed in the posterior end of the worm along the primary body axis in evaginated cysticerci. These findings indicated that wnt4 may take part in the process of cysticerci evagination and play a role in scolex/bladder development of cysticerci of T. solium.
Assuntos
Cysticercus/genética , DNA de Helmintos/genética , Taenia solium/genética , Proteína Wnt4/genética , Animais , Sequência de Bases , Cisticercose/patologia , Cysticercus/enzimologia , Regulação da Expressão Gênica , Humanos , Hibridização In Situ , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Sus scrofa , Suínos , Doenças dos Suínos , Taenia solium/embriologia , Taenia solium/enzimologiaRESUMO
Foot-and-mouth disease (FMD) is a highly contagious and economically important disease of cloven-hoofed animals that hampers trade and production. To ensure effective infection, the foot-and-mouth disease virus (FMDV) evades host antiviral pathways in different ways. Although the effect of histone deacetylase 5 (HDAC5) on the innate immune response has previously been documented, the precise molecular mechanism underlying HDAC5-mediated FMDV infection is not yet clearly understood. In this study, we found that silencing or knockout of HDAC5 promoted FMDV replication, whereas HDAC5 overexpression significantly inhibited FMDV propagation. IFN-ß and IFN-stimulated response element (ISRE) activity was strongly activated through the overexpression of HDAC5. The silencing and knockout of HDAC5 led to an increase in viral replication, which was evident by decreased IFN-ß, ISG15, and ISG56 production, as well as a noticeable reduction in IRF3 phosphorylation. Moreover, the results showed that the FMDV capsid protein VP1 targets HDAC5 and facilitates its degradation via the proteasomal pathway. In conclusion, this study highlights that HDAC5 acts as a positive modulator of IFN-ß production during viral infection, while FMDV capsid protein VP1 antagonizes the HDAC5-mediated antiviral immune response by degrading HDAC5 to facilitate viral replication.
Assuntos
Vírus da Febre Aftosa , Febre Aftosa , Interferon Tipo I , Animais , Proteínas do Capsídeo/metabolismo , Transdução de Sinais , Febre Aftosa/metabolismo , Imunidade Inata , Interferon Tipo I/metabolismoRESUMO
Orf virus (ORFV) is an enveloped virus with a double-stranded DNA genome, causing a contagious pustular dermatitis, mainly in goats and sheep. In this study, two strains of ORFV were isolated from sheep and goat samples in Xinjiang and Shaanxi, China. The B2L and virus interferon resistance (VIR) genes of these two isolates were sequenced and analyzed after PCR amplification. Phylogenetic analysis showed that the two isolates clustered with other ORFV strains but were separated into different subgroups. The Xinjiang strain shared the highest homology with the Gansu strain, whereas the Shaanxi strain shared higher homology with the Taiwan and Hubei strains. This is the first report of the molecular characterization of ORFV in Northwest China, and it provides new information on the genotyping of the causative agents responsible for contagious ecthyma dermatitis outbreaks in China.
Assuntos
Ectima Contagioso/virologia , Vírus do Orf/classificação , Vírus do Orf/genética , Filogenia , Proteínas Virais/genética , Animais , China , Análise por Conglomerados , DNA Viral/química , DNA Viral/genética , Genótipo , Cabras , Dados de Sequência Molecular , Vírus do Orf/isolamento & purificação , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , OvinosRESUMO
BACKGROUND: Goatpox is an economically important disease in goat and sheep-producing areas of the world. Many vaccine strategies developed to control the disease are not yet completely successful. Hairpin expression vectors have been used to induce gene silencing in a large number of studies on viruses. However, none of these studies has been attempted to study GTPV. In the interest of exploiting improved methods to control goat pox, it is participated that RNAi may provide effective protection against GTPV. In this study we show the suppression of Goatpox virus (GTPV) replication via knockdown of virion core protein using RNA interference. RESULTS: Four short interfering RNA (siRNA) sequences (siRNA-61, siRNA-70, siRNA-165 and siRNA-296) against a region of GTPV ORF095 were selected. Sense and antisense siRNA-encoding sequences separated by a hairpin loop sequence were designed as short hairpin RNA (shRNA) expression cassettes under the control of a human U6 promoter. ORF095 amplicon was generated using PCR, and then cloned into pEGFP-N1 vector, named as p095/EGFP. p095/EGFP and each of the siRNA expression cassettes (p61, p70, p165 and p296) were co-transfected into BHK-21 cells. Fluorescence detection, flow cytometric analysis, retro transcription PCR (RT-PCR) and real time PCR were used to check the efficiency of RNAi. The results showed that the ORF095-specific siRNA-70 effectively down-regulated the expression of ORF095. When Vero cells were transfected with shRNA expression vectors (p61/GFP, p70/GFP, p165/GFP and p296/GFP) and then infected with GTPV, GTPV-ORF095-70 was found to be the most effective inhibition site in decreasing cytopathic effect (CPE) induced by GTPV. The results presented here indicated that DNA-based siRNA could effectively inhibit the replication of GTPV (approximately 463. 5-fold reduction of viral titers) on Vero cells. CONCLUSIONS: This study demonstrates that vector-based shRNA methodology can effectively inhibit GTPV replication on Vero cells. Simultaneously, this work represents a strategy for controlling goatpox, potentially facilitating new experimental approaches in the analysis of both viral and cellular gene functions during of GTPV infection.
Assuntos
Capripoxvirus/fisiologia , Interferência de RNA , Proteínas Virais/genética , Replicação Viral , Animais , Antivirais/farmacologia , Produtos Biológicos/farmacologia , Capripoxvirus/genética , Capripoxvirus/crescimento & desenvolvimento , Chlorocebus aethiops , RNA Interferente Pequeno/farmacologia , Células Vero , Proteínas Virais/biossínteseRESUMO
BACKGROUND: Porcine reproductive and respitatory syndrome virus (PRRSV) is a recently emerged pathogen and severely affects swine populations worldwide. The replication of PRRSV is tightly controlled by viral gene expression and the codon usage of translation initiation region within each gene could potentially regulate the translation rate. Therefore, a better understanding of the codon usage pattern of the initiation translation region would shed light on the regulation of PRRSV gene expression. RESULTS: In this study, the codon usage in the translation initiation region and in the whole coding sequence was compared in PRRSV ORF1a and ORFs2-7. To investigate the potential role of codon usage in affecting the translation initiation rate, we established a codon usage model for PRRSV translation initiation region. We observed that some non-preferential codons are preferentially used in the translation initiation region in particular ORFs. Although some positions vary with codons, they intend to use codons with negative CUB. Furthermore, our model of codon usage showed that the conserved pattern of CUB is not directly consensus with the conserved sequence, but shaped under the translation selection. CONCLUSIONS: The non-variation pattern with negative CUB in the PRRSV translation initiation region scanned by ribosomes is considered the rate-limiting step in the translation process.
Assuntos
Códon , Regulação Viral da Expressão Gênica , Genoma Viral , Iniciação Traducional da Cadeia Peptídica , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Fases de Leitura Aberta , Ribossomos/metabolismo , Proteínas Virais/biossínteseRESUMO
Peste des Petits Ruminant (PPR) is an infectious viral disease of small ruminants caused by PPR virus. Although goat and sheep are the primary hosts of PPR, studies have continuously reported the prevalence of circulating antibodies in large ruminants, which could bring a potential challenge to effectively control and eradicate PPR. In Nepal, seroprevalence of PPRV antibodies in cattle have not been monitored yet. To address this, a total of 255 cattle sera were collected from Rupandehi, Banke, Bara and Chitwan districts of Nepal where outbreak of PPR in small ruminants was reported previously. The sera samples were tested by competitive ELISA and the result indicated the prevalence of 5.88% PPRV antibodies in cattle which indicates the exposure of cattle to PPR virus. To make the disease control program effective, intensive monitoring of both domestic and wild animals is very important.
Assuntos
Doenças dos Bovinos/epidemiologia , Peste dos Pequenos Ruminantes/epidemiologia , Vírus da Peste dos Pequenos Ruminantes/isolamento & purificação , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Nepal/epidemiologia , Peste dos Pequenos Ruminantes/microbiologia , Prevalência , Estudos SoroepidemiológicosRESUMO
Peste des petits ruminant virus (PPRV) causes a highly contagious disease in small ruminants. The molecular mechanism of PPRV replication and its interactions with hosts are poorly studied. In other paramyxoviruses, the viral phosphoprotein (P) has been associated with multiple functions for key biological processes such as the regulation of transcription, translation, and the control of cell cycle. Phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF2α) is an important process for gene regulation in host cells under stress, including viral infection. In the present study, molecular mechanisms associated with PPRV replication and viral interaction with host cells were investigated. We describe the ability of PPRV to dephosphorylate eIF2α and the potential of PPRV P protein to induce the host cellular growth arrest DNA damage protein (GADD34), which is known to be associated with eIF2α dephosphorylation. Furthermore, we observed that PPRV P protein alone could block PERK/eIF2α phosphorylation. We speculate that PPRV exploits eIF2α dephosphorylation to facilitate viral replication and that PPRV P protein is involved in this molecular mechanism. This work provides new insights into further understanding PPRV pathobiology and its viral/host interactions.
RESUMO
Peste des petits ruminant (PPR) is an economically important severe viral disease of small ruminants that affects primarily the respiratory and digestive tract. Specific detection of the PPR virus (PPRV) antigen plays an important role in the disease control and eradication program. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant goat signaling lymphocyte activation molecule (SLAM) as the capture ligand was successfully developed for the detection of the PPRV antigen (PPRV SLAM-iELISA). The assay was highly specific for PPRV with no cross-reactions among foot and mouth disease virus, Orf virus, sheep pox virus, and goat pox virus and had a sensitivity with a detection limit of 1.56 × 101 TCID50/reaction (50 µl). Assessment of 136 samples showed that the developed PPRV SLAM-iELISA was well correlated with real-time RT-qPCR assays and commercially available sandwich ELISA for detection of PPRV and showed relative sensitivity and specificity of 93.75 and 100.83%, respectively. These results suggest that the developed PPRV SLAM-iELISA is suitable for specific detection of the PPRV antigen. This study demonstrated for the first time that the goat SLAM, the cellular receptor for PPRV, can be used for the development of a diagnostic method for the detection of PPRV.
RESUMO
Peste des petits ruminants (PPR) is a severe respiratory and digestive tract disease of domestic small ruminants caused by PPR virus (PPRV) of the genus Morbillivirus. Although the primary hosts of PPRV are goats and sheep, the host range of PPRV has been continuously expanding and reported to infect various animal hosts over the last decades, which could bring a potential challenge to effectively control and eradicate PPR globally. In this review, we focused on current knowledge about host expansion and interspecies infection of PPRV and discussed the potential mechanisms involved.
RESUMO
Peste des Petits Ruminant (PPR) is an important transboundary, OIE-listed contagious viral disease of primarily sheep and goats caused by the PPR virus (PPRV), which belongs to the genus Morbillivirus of the family Paramyxoviridae. The mortality rate is 90-100%, and the morbidity rate may reach up to 100%. PPR is considered economically important as it decreases the production and productivity of livestock. In many endemic poor countries, it has remained an obstacle to the development of sustainable agriculture. Hence, proper control measures have become a necessity to prevent its rapid spread across the world. For this, detailed information on the pathogenesis of the virus and the virus host interaction through cellular receptors needs to be understood clearly. Presently, two cellular receptors; signaling lymphocyte activation molecule (SLAM) and Nectin-4 are known for PPRV. However, extensive information on virus interactions with these receptors and their impact on host immune response is still required. Hence, a thorough understanding of PPRV receptors and the mechanism involved in the induction of immunosuppression is crucial for controlling PPR. In this review, we discuss PPRV cellular receptors, viral host interaction with cellular receptors, and immunosuppression induced by the virus with reference to other Morbilliviruses.
Assuntos
Interações Hospedeiro-Patógeno , Peste dos Pequenos Ruminantes/metabolismo , Peste dos Pequenos Ruminantes/virologia , Vírus da Peste dos Pequenos Ruminantes/fisiologia , Receptores de Superfície Celular/metabolismo , Receptores Virais/metabolismo , Animais , Genoma Viral , Genômica/métodos , Especificidade de Hospedeiro , Interações Hospedeiro-Patógeno/imunologia , Imunidade , Peste dos Pequenos Ruminantes/imunologia , Ligação Proteica , Receptores de Superfície Celular/química , Receptores Virais/química , Transdução de Sinais , Relação Estrutura-Atividade , Proteínas Virais/química , Proteínas Virais/metabolismoRESUMO
Due to the migration or transboundary spread of domestic and wild animals, peste des petits ruminants virus posed a high potential threat to them. In this study, we initially detected that a class of animal named Procapra przewalskii was infected with peste des petits ruminants virus (PPRV ChinaGS2018) in Gansu province. According to phylogenetic relationships analysis, we found that ChinaGS2018 comprised of 15,954 nucleotides and was classified into IV genotypes. In addition, indirect immunofluorescence assay (IFA) showed that ChinaGS2018 could infect isolated primary goat tracheal epithelium cells (GTC). Comparing with full-length genome sequences revealed that ChinaGS2018 strain has high identity to the reference complete genomes (87.16-99.55%) at the nucleotide level. Multiple sequence alignment showed that F protein has the highest identity of 99.8%, and H protein has the highest nucleotide substitution ratio. Our study also suggested this strain may be transmitted from Xinjiang, China. Along with the migratory of Procapraprzewalskii, this wild ruminant infected with PPRV can pose a huge threat to other wild ruminants and domestic ones. This is the first report describing infected with PPRV which will provide insights into the epidemiology and pathogenesis of this important virus.
Assuntos
Peste dos Pequenos Ruminantes/epidemiologia , Peste dos Pequenos Ruminantes/virologia , Vírus da Peste dos Pequenos Ruminantes/genética , Ruminantes/virologia , Animais , Animais Selvagens , Autopsia , China/epidemiologia , Biologia Computacional/métodos , Evolução Molecular , Genoma Viral , Genômica/métodos , Peste dos Pequenos Ruminantes/diagnóstico , Vírus da Peste dos Pequenos Ruminantes/classificação , Filogenia , Prevalência , Vigilância em Saúde PúblicaRESUMO
Peste des petits ruminants (PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus (PPRV) pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals, establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.
Assuntos
Peste dos Pequenos Ruminantes/prevenção & controle , Vírus da Peste dos Pequenos Ruminantes/genética , Vírus de RNA/genética , Genética Reversa , Animais , Surtos de Doenças/veterinária , Genoma Viral , Filogenia , RNA ViralRESUMO
OBJECTIVE: To express the TSO45W-4BX of Taenia solium in combination with CD58 as a molecular adjuvant for improving the protective efficacy of the TSO45W-4BX recombinant vaccine. METHODS: TSO45W-4BX and porcine CD58 genes were amplified by PCR, using recombinant plasmids pGEM-4B and pGEM-CD58 as template respectively. The CD58 fragment was inserted into the recombinant plasmid pGEX-4T-1 with directly ligated TSO45W-4BX. The transformant was induced with IPTG and followed by identifying the integrity of the recombinant containing TS045W-4BX and porcine CD58 with PCR and sequencing. The products were analyzed by SDS-PAGE and Western blotting. RESULTS: The expression products of Mr 69,000 GST-4BX/CD58 and Mr 41,000 GST-4BX were present mainly in the form of inclusion bodies and soluble substance respectively, and both were recognized by sera of cysticercosis patients. CONCLUSIONS: The TSO45W-4BX co-expressed with porcine CD58 conserves its immune reactivity.
Assuntos
Antígenos de Protozoários/biossíntese , Antígenos CD58/biossíntese , Cisticercose/sangue , Cisticercose/parasitologia , Escherichia coli/metabolismo , Taenia solium/imunologia , Animais , Antígenos de Protozoários/genética , Escherichia coli/genética , Humanos , Plasmídeos , Reação em Cadeia da Polimerase , Suínos , Taenia solium/genéticaRESUMO
In order to prove the adjuvant effect of CpG DNA recombinant plasmid, the total antibodies and their IgG2a subtype induced by antigen of Cysticercus cellulosae, and content of IL-4 and IFN-gamma secreted from splenic cell of mouse immunized were measured. The recombinant plasmids showed an adjuvant effect, and CpG2 was the best adjuvant among the plasmids. It is proved that the CpG DNA possesses a synergistic effect with AI(OH)3 and 206 adjuvant, and is an effective Th1 type adjuvant in mice.
Assuntos
Adjuvantes Imunológicos/farmacologia , Antígenos de Helmintos/imunologia , DNA/imunologia , Animais , Ilhas de CpG/genética , Cisticercose/sangue , Cisticercose/imunologia , Cisticercose/prevenção & controle , Cysticercus/imunologia , Feminino , Imunização , Interferon gama/metabolismo , Interleucina-2/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Plasmídeos/genética , Distribuição Aleatória , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismoRESUMO
Peste des Petits Ruminants (PPR) is an acute, highly contagious, and febrile viral disease that affects both domestic and wild small ruminants. The disease has become a major obstacle to the development of sustainable Agriculture. Hemagglutinin (H), the envelope glycoprotein of Peste des Petits Ruminants Virus (PPRV), plays a crucial role in regulating viral adsorption and entry, thus determining pathogenicity, and release of newly produced viral particles. In order to accurately understand the epidemic of the disease and the interactions between the virus and host, we launch the work. Here, we examined H gene from all four lineages of the PPRV to investigate evolutionary and epidemiologic dynamics of PPRV by the Bayesian method. In addition, we predicted positive selection sites due to selective pressures. Finally, we studied the interaction between H protein and SLAM receptor based on homology model of the complex. Phylogenetic analysis suggested that H gene can also be used to investigate evolutionary and epidemiologic dynamics of PPRV. Positive selection analysis identified four positive selection sites in H gene, in which only one common site (aa246) was detected by two methods, suggesting strong operation structural and/or functional constraint of changes on the H protein. This target site may be of interest for future mutagenesis studies. The results of homology modeling showed PPRVHv-shSLAM binding interface and MVH-maSLAM binding interface were consistent, wherein the groove in the B4 blade and B5 of the head domain of PPRVHv bound to the AGFCC' ß-sheets of the membrane-distal ectodomain of shSLAM. The binding regions could provide insight on the nature of the protein for epitope vaccine design, novel drug discovery, and rational drug design against PPRV.
Assuntos
Evolução Molecular , Hemaglutininas Virais/genética , Vírus da Peste dos Pequenos Ruminantes/genética , Genes Virais , FilogeniaRESUMO
Taenia saginata, Taenia solium and Taenia asiatica (beef, pork and Asian tapeworms, respectively) are parasitic flatworms of major public health and food safety importance. Among them, T. asiatica is a newly recognized species that split from T. saginata via an intermediate host switch â¼1.14 Myr ago. Here we report the 169- and 168-Mb draft genomes of T. saginata and T. asiatica. Comparative analysis reveals that high rates of gene duplications and functional diversifications might have partially driven the divergence between T. asiatica and T. saginata. We observe accelerated evolutionary rates, adaptive evolutions in homeostasis regulation, tegument maintenance and lipid uptakes, and differential/specialized gene family expansions in T. asiatica that may favour its hepatotropism in the new intermediate host. We also identify potential targets for developing diagnostic or intervention tools against human tapeworms. These data provide new insights into the evolution of Taenia parasites, particularly the recent speciation of T. asiatica.