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ABSTRACT: BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is largely responsible for heavy economic losses in the swine industry worldwide because of its high mutation rate and subsequent emergence of virulent strains. However, the immunological and pathological responses of pigs to PRRSV strains with different virulence have not been completely elucidated. METHODS: Twenty-four piglets were divided into 4 groups (n = 6 each) and inoculated with highly pathogenic PRRSV isolate BB0907 (HP), low pathogenic PRRSV NT0801 (LP), LP derivative strain NT0801-F70 (LP-der), and DMEM medium (control), respectively. The changes in TLR2, 3, 7, and 8 gene expression and TNF-alpha, IL-1beta, IL-6, IFN-gamma, and IL-10 secretion were evaluated using real-time PCR and ELISA at 6, 9, and 15 days post inoculation (d.p.i.). The cytokine levels were evaluated in the supernatants of porcine alveolar macrophages (PAMs) and peripheral blood mononuclear cells (PBMCs) following stimulation with LTA, poly(I:C), CL097, and PRRSV individually. RESULTS: HP caused more severe clinical signs and pathological lesions in swine than LP and LP-der had almost no virulence compared with LP. The serum levels of IL-1beta, IL-6, TNF-alpha, and IFN-gamma were increased in HP-infected piglets, which were greater than in those infected with LP or LP-der. The mRNA levels of TLR3, 7, and 8 were significantly up-regulated in PAMs in HP-infected pigs compared to those in groups LP and LP-der. Furthermore, TNF-alpha and IL-1beta secretion in PAMs from group LP was statistically greater than those from the control group after stimulation with either poly(I:C) or CL097. Meanwhile, TNF-alpha, IL-1beta, and IL-6 levels in CL097-stimulated PBMCs from HP-infected pigs were markedly higher than those from the LP- and LP-der-infected groups. CONCLUSIONS: We found that HP was a stronger inducer of TLR 3, 7, and 8 expression and IL-1beta, IL-6, TNF-alpha, and IFN-gamma production compared to LP and LP-der. HP enhanced production of TNF-alpha, IL-1beta, and IL-6 in PBMCs following CL097-stimulation more than LP and LP-der, whereas LP enhanced the secretion of TNF-alpha and IL-1beta in poly(I:C)- and CL097-stimulated PAMs. Our data regarding cellular reactivity to different isolates should be useful in the development of more efficacious vaccines.
Virology Journal 04/2013; 10(1):135. · 2.34 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) is mainly responsible for the heavy economic losses in pig industry in the world. Current vaccination strategies provide only a limited protection. Previous studies have demonstrated the immunostimulatory adjuvant effects of Toll-like receptor (TLR) ligands, synthetic double-stranded RNA polyriboinosinic polyribocytidylic [poly(I:C)], lipoteichoic acid (LTA) and CL097 in humans and animals. To study the effects of these compounds on the induction of PRRSV-specific immune responses, mice were immunized subcutaneously with killed virus (KV) antigens incorporating pairs of TLR ligands. It was found that poly(I:C) and CL097 induced the higher IFN-γ levels and PRRSV-specific antibodies, comparing with that KV with or without LTA in mice. Piglets were vaccinated with the KV mixed with poly(I:C) or CL097 and the protective effects of the vaccination were evaluated. The results showed that PRRSV-specific antibodies and T lymphocyte proliferation levels in KV mixed with poly(I:C) or CL097 groups were higher than those in KV group. Following challenge with PRRSV, pigs inoculated with KV mixed with poly(I:C) or CL097 showed lighter clinical signs, lower viremia and less pathological lesion of lungs, as compared to those of KV and challenge control groups. It indicated that co-administration of poly(I:C) and CL097 with killed PRRSV vaccine conferred higher protection against PRRSV challenge. TLR3 and TLR7/8 ligands are promising adjuvant candidates for the development of novel vaccines against PRRSV.
Veterinary Microbiology 02/2013; · 3.33 Impact Factor
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ABSTRACT: Porcine circovirus type-2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases and has 4 main ORFs, ORF1 (Rep gene), ORF2 (Cap gene), ORF3 within ORF1, and ORF4, which is overlapped with ORF3, and 1 origin of replication located between ORF1 and ORF2. The chimeric PCV1-2, containing the PCV2 capsid, PCV1 rep, and Ori genes, is attenuated in pigs. In order to verify the role of the Rep gene or Ori in the virulence of PCV2, 3 chimeric viruses [PCV2b-Ori1 (PCV1 Ori gene cloned into the backbone of PCV2b), PCV2b-rep1 (PCV1 Rep gene cloned into the backbone of PCV2b), and PCV2b-rep1-Ori1 (PCV1 Rep and Ori genes cloned into the backbone of PCV2b)] and 2 wild-type recombinant PCV2b and PCV1 were constructed and identified. The experimental results in piglets showed that clinical symptoms, viremia, viral load, lesions in lymphoid and lung tissues, and IL-10 and TNF-α expression levels in PBMCs in the PCV2b-rep1-Ori1 and PCV2b-rep1 groups were significantly decreased, compared to PCV2-infected piglets. Meanwhile, histological lesions of lymphoid and lung tissues, viral loads in lymphoid tissues, viremia, and TNF-α expression in PBMCs were not significantly different between groups PCV2b-Ori1 and PCV2b, suggesting that the Rep gene (ORF1) likely contributes to viral pathogenicity in vivo.
Virus Research 02/2013; · 2.94 Impact Factor
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ABSTRACT: In order to obtain a virus-like particles (VLPs) vaccine both for porcine circovirus type 2 (PCV2) prevention and growth-promotion, somatostatin (SS) gene was fused to the 3'-terminal of ORF2 gene of PCV2 with PCR, and a recombinant baculovirus (rAc-Cap-SS) was constructed. The expression of fusion protein Cap-SS (rCap-SS) with molecular weight of approximately 32kDa was identified by Western blot and indirect immunofluorescence assay in Sf9 cells. The self-assembled VLPs were observed under electron microscopy, which being morphologically similar to the recombinant Cap protein (rCap) expressed in the same baculovirus expressing system. Ninety four-week-old mice were immunized with the recombinant proteins twice. The results showed that mice immunized with rCap-SS protein developed antibody against Cap, which levels being similar to those immunized with rCap protein. The body weight gain and anti-SS antibody in rCap-SS group was higher than those of rCap and negative control groups during 28 and 42 days post inoculation (dpi). Furthermore, twenty 28-day-old piglets were vaccinated twice subcutaneously with the recombinant proteins. The results indicated that PCV2-specific antibody could be induced after vaccination with rCap-SS or rCap protein. Anti-SS antibody could be induced after rCap-SS vaccination and was higher than other groups at 14 and 28 dpi. The level of somatostatin concentration in the blood of pigs in rCap-SS group was significantly decreased at 14 dpi than other groups (P<0.05). The relative daily weight gain (RDWG) of pigs in rCap-SS group was obviously higher than that in other groups at 28 dpi. After challenge with PCV2, pigs in the vaccinated groups had no clearly clinical signs, and the RDWG was significantly higher than that in the challenge control group (CC) (P<0.05). The pathological lesions, viremia and viral load presented in the vaccinated groups were milder than those in challenge control group. It suggested that the recombinant porcine circovirus-like particles displaying somatostatin might be a novel subunit vaccine candidate for preventing PMWS and promoting pig growth.
Veterinary Microbiology 12/2012; · 3.33 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome (PRRS) is characterized by a delayed and defective adaptive immune response. The viral nonstructural protein 1 (NSP1) of the PRRS virus (PRRSV) is able to suppress the type I interferon (IFN) response in vitro. In this study, recombinant adenoviruses (rAds) expressing NSP1 (rAd-NSP1), glycoprotein 5 (GP5) (rAd-GP5), and the NSP1-GP5 fusion protein (rAd-NSP1-GP5) were constructed, and the effect of NSP1 on immune responses was investigated in pigs. Pigs inoculated with rAd-NSP1 or rAd-NSP1-GP5 had significantly lower levels of IFN-γ and higher levels of the immunosuppressive cytokine IL-10 than pigs inoculated with rAd-GP5, wild-type adenovirus, or cell culture medium alone. The antibody response to vaccination against classic swine fever virus (CSFV) was significantly decreased by inoculation of NSP1 7 d after CSFV vaccination in pigs. Thus, NSP1-mediated immune suppression may play an important role in PRRSV pathogenesis.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 10/2012; 76(4):255-60. · 0.94 Impact Factor
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ABSTRACT: Interactions between host factors and the viral protein play important roles in host adaptation and regulation of virus replication. Poly(A)-binding protein (PABP), a host cellular protein that enhances translational efficiency by circularizing mRNAs, was identified by yeast two-hybrid screening as a cellular partner for PRRSV nucleocapsid (N) protein in porcine alveolar macrophages. The specific interaction of PRRSV N protein with PABP was confirmed in infected cells by co-immunoprecipitation and in vitro by GST pull-down assay. We showed by confocal microscopy that the PABP co-localized with the PRRSV N protein. Using a series of deletion mutants, the interactive domain of N protein with PABP was mapped to a region of amino acids 52-69. For PABP, C-terminal half, which interestingly interacts other translation regulators, was determined to be the domain interactive with N protein. Short hairpin RNA (shRNA)-mediated silencing of PABP in cells resulted in significantly reduced PRRSV RNA synthesis, viral encoded protein expression and viral titer. Overall, the results presented here point toward an important role for PABP in regulating PRRSV replication.
Antiviral research 09/2012; · 3.61 Impact Factor
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ABSTRACT: Encephalomyocarditis virus (EMCV) infection can cause myocarditis and sudden death in pre-weaned piglets and severe reproductive failure in sows. There are no specific antiviral drugs for the treatment of the virus infection. In this study, four recombinant adenoviruses expressing small interfering RNAs (siRNAs) targeting to 1D or 3AB protein genes of EMCV were constructed and their inhibition efficiency on the replication of EMCV was evaluated in both Marc-145 cells and mice. The results showed that the rAd5 expressing siRNAs (rAd5) could inhibit EMCV replication in Marc-145 cells in protein and mRNA levels, as well as the virus yield by approximately 100-1000 times. And the inhibition of viral replication was sustained for 72h and dose-dependent. Animal experiment results showed that EMCV VP1 mRNA level in the brain of mice in the rAd5 groups were obviously lower than those in rAd-G1 and challenge control groups. The virus yields in rAd-1D-2 and rAd-3AB-1 groups were markedly decreased by more than 90.0%. The survival rates of mice in rAd-1D-2 group were significantly higher than those in challenge control groups. Furthermore, the survival mice only showed minor microscopic lesions in brain and minor edema of nerve cell, which was obviously slighter than those in challenge control groups. These results indicated that siRNAs mediated by the adenovirus could provide protective efficacy against EMCV challenge in mice. It might provide a potential strategy for combating EMCV.
Journal of virological methods 07/2012; 185(2):204-12. · 2.13 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome (PRRS) has devastated the pig industry worldwide for almost 25 years, and its virus (PRRSV) preferentially infects and replicates in pulmonary alveolar macrophages (PAMs). To discover cellular protein responses in PRRSV-infected PAMs, two-dimensional liquid chromatography-tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling was employed to quantitatively identify the differentially expressed proteins between the PRRSV-infected groups and the controls. A total of 160 cellular proteins in PAMs that were significantly altered post-infection were identified. These differentially expressed proteins are related to the biological processes of virus binding, cell structure, signal transduction, cell adhesion, etc., and their interactions. This is the first report that analyzed the cellular protein profile of PRRSV-infected PAMs using iTRAQ technology, and this data provides important information to help understand the host response to PRRSV and to define the cellular requirements for the underlying mechanism of PRRSV replication and pathogenesis.
Journal of Proteome Research 04/2012; 11(5):2890-903. · 5.11 Impact Factor
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ABSTRACT: Encephalomyocarditis virus (EMCV) can infect many host species and cause acute myocarditis and sudden death in preweaned piglets. In this study, an EMCV strain (NJ08) was isolated from newborn pigs with clinical signs on a pig farm in mideastern China. It was identified by indirect immunofluorescence assay and reverse-transcription polymerase chain reaction. Experiments showed that the isolate could cause severe clinical symptoms and pathological changes in mice but no obvious clinical and pathological changes in commercial piglets. Complete genomic sequencing showed that the NJ08 strain was 78.3% to 100% identical with other isolates in regions coding for various proteins. Phylogenetic analysis showed that the NJ08 isolate belonged to subgroup Ia. This study confirmed that an EMCV isolate from pigs could be fatal to mice and provided new epidemiologic data on EMCV in China.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 04/2012; 76(2):157-60. · 0.94 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) was divided into North American and European genotypes. NSP1 was an important non-structural protein of PRRSV, which was auto-cleaved from the replicase polyprotein into NSP1α and NSP1β subunits and played an important role in the immune suppression. In this study, six monoclonal antibodies (MAbs) against the recombinant PRRSV NSP1, expressed in Escherichia coli system, were screened out and identified. Western blot and IFA results indicated that 4 out of 6 MAbs recognized the recombinant NSP1α and 2 MAbs recognized NSP1β. Epitope mapping results indicated that MAb 4H2 recognized the linear epitopes E(54)EPLRW(59) in NSP1α, MAbs (2G5, 3E11 and 4D4) recognized the epitopes H(157)VLTNLP(163) in NSP1α, and MAbs 3C7 and 1H7 reacted with the epitopes 185aa to 232aa in NSP1β. Protein sequence alignment of NSP1 indicated E(54)EPLRW(59) was conserved in all North American PRRSV strains, whereas European type strains has variable amino acids in this region. The epitope H(157)VLTNLP(163) was relatively conserved among all PRRSV strains, except for a L162→S162 change in European type strains. The epitope 185-232aa was variable among North American PRRSV strains. These results may facilitate future investigations into the function of NSP1 of PRRSV and diagnostic methods for PRRSV infection.
Veterinary Microbiology 03/2012; 155(2-4):220-9. · 3.33 Impact Factor
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ABSTRACT: Aleutian mink disease virus (AMDV) is a parvovirus that causes an immune complex mediated disease in minks. To understand the genetic characterization of AMDV in China, the genomic sequences of three isolates, ADV-LN1, ADV-LN2, and ADV-LN3, from different farms in the Northern China were analyzed. The results showed that the lengths of genomic sequences of three isolates were 4,543, 4,566, and 4,566 bp, respectively. They shared only 95.5-96.3 % nucleotide identity with each other. The nucleotide and amino acid homology of genome sequence between the Chinese isolates and European or American strains (ADV-G, ADV-Utah1, and ADV-SL3) were 92.4-95.0 % and 92.1-93.8 %, respectively. The amino acid substitutions randomly distributed in the genome, especially NS gene. ADV-LN1 strain had a 9-amino-acid deletion at amino acid positions 70 and 72-79 in the VP1 gene, comparing with ADV-G strain; ADV-LN2 and ADV-LN3 strains had 1-amino-acid deletion at amino acid positions 70 in the VP1. Some potential glycosylation site mutations in VP and NS genes were also observed. Phylogenetic analysis results showed that the three strains belonged to two different branches based on the complete coding sequence of VP2 gene. However, they all were in the same group together with the strains from United States based on the NS1 sequence. It indicated that Chinese AMDV isolates had genetic diversity. The origin of the ancestors of the Chinese AMDV strains might be associated with the American strains.
Virus Genes 03/2012; 45(1):24-30. · 1.85 Impact Factor
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ABSTRACT: Encephalomyocarditis virus (EMCV) could infect many host species and cause acute myocarditis and sudden death in pre-weaned piglets. It was necessary to develop new antiviral strategies for the treatment of the virus infection. Here, four plasmids expressing shRNA (small hairpin RNA) targeted to 1D or 3AB protein genes of EMCV were constructed and their inhibition efficiency on the replication of EMCV was evaluated in both BHK21 cells and mice. The results showed that three out of those four shRNA constructs could significantly inhibit EMCV replication in BHK21 cells on the levels of viral RNA and protein. Moreover, it was found that the shRNAs could suppress significantly the load of EMCV in the brain tissue of the mice pretreated with the constructs for 6-24 h. The clinical signs and pathological lesions of the mice in the groups inoculated with the shRNA constructed were milder obviously, compared with those in pSUPER-mN3 and challenge control groups. The survival rates of mice inoculated with pSUPER-3AB-1, pSUPER-3AB-2, and pSUPER-1D-1 for 12 h was 100, 80, and 40%, respectively, while, in the control groups it was only 20%. It indicated that the vector-based shRNA targeting to 3AB and 1D genes might be a potential anti-EMCV strategy.
Virus Genes 11/2011; 44(2):183-90. · 1.85 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the important pathogens causing serious economic losses to swine industry worldwide. PRRSV is genetically and pathologically heterogenous. PRRSV NT0801 strain was isolated in a pig farm with clinical signs and had high pathogenesis in piglets. But its NSP2 gene did not have 30 amino acids deletion as highly pathogenic JXA1 strain. To elucidate the genetic characteristics of PRRSV NT0801 strain, the full-length genome of NT0801 isolate was sequenced and analyzed. The results showed that the genome of PRRSV NT0801 was 15439bp in length, including 29nt Poly(A) tail. Compared with the highly pathogenic JXA1 strain, it had the nucleotide sequence identity of 96.7%, amino acid sequence homology of 97.2% and 98.5% in GP3 and GP5, respectively. Phylogenetic analysis indicated that NT0801 isolate was located between the traditional strain and the highly pathogenic strain. But no obvious recombination signal was observed, compared with other PRRSV isolates with different virulence. The alignment of amino acid sequence of NT0801 with other PRRSV isolates demonstrated that three out of nine sites, being consistent with the highly pathogenic strain, were different from those in highly pathogenic while same as those in traditional strains and JXA1 vaccine strain. And one out of 9 sites was same as that of JXA1 vaccine strain exclusively, two out of 9 sites were different from all the strains. These results indicated that PRRSV NT0801 strain is closely related to highly pathogenic PRRSV, although there has no 30 amino acids deletions in NSP2 region. The epidemic PRRSV strains variation results from the gene mutation. It should be useful for studying on the virulence genes located in different ORFs of PRRSV in the future.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 11/2011; 27(6):542-8.
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ABSTRACT: Haemophilus parasuis is the etiological agent of Glässer's disease characterized by fibrinous polyserositis, polyarthritis, and meningitis in young pigs. But it is difficult to develop universal serological diagnostic tools and effective vaccines against this disease because of the serovar diversity of the isolates. In this study, enterobacterial repetitive intergenic consensus-polymerase chain reaction, were performed to investigate the gene profile of 111 isolates of H. parasuis from China. And a specific common gene of H. parasuis was cloned and identified as the outer-membrane protein (OMP) P2 gene. Sequencing results of OMP P2 genes of 22 isolates showed that they had high homology and could be divided into 2 genetic types. Moreover, the OMPP2 protein was expressed in Escherichia coli expressing system. And the purified recombinant protein provided partial protection against H. parasuis infection in mice. It suggested the OMP P2 was an immunogenic protein and had great potential to serve as a vaccine and diagnostic antigen.
Research in Veterinary Science 09/2011; 93(2):736-42. · 1.65 Impact Factor
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ABSTRACT: Infectious bronchitis virus (IBV) causes tremendous economic losses associated with production inefficiencies and mortality in poultry industry worldwide. In the present report, the recombinant adenoviruses expressing chicken granulocyte-macrophage colony stimulating factor (GM-CSF) and S1 gene of nephropathogenic IBV were constructed and characterized. Then, the immunological efficacy and protection against homologous IBV challenge were assessed in specific pathogen free (SPF) chickens. The results showed that the chickens vaccinated in ovo with rAd-S1, rAd-GM-S1 (GM-CSF fused with S1 using glycine linkers) and rAd-GM-CSF plus rAd-S1 (co-administered) developed specific anti-IBV HI antibodies. Moreover, the fusion of the GM-CSF markedly increased spleen cell proliferation and IFN-γ production while mild increased in IL-4 production, which demonstrated the enhancement of cell-mediated immune responses. Following challenge with IBV, the chickens in the group vaccinated with rAd-S1 fused or co-administered with GM-CSF had fewer nephropathic lesions and showed 100% protection as compared to that of rAd-S1 alone which showed 70% protection. It indicated that the single dose in ovo vaccination of the GM-CSF fused or co-administered with S1 of IBV could enhance significantly the humoral, cellular immune responses and provide complete protection against nephropathogenic IBV challenge. This finding may provide basic information for effective in ovo vaccines design against IBV.
Veterinary Microbiology 02/2011; 148(1):8-17. · 3.33 Impact Factor
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ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) has recently caused heavy economic losses in swine industry worldwide. Current vaccination strategies only provide a limited protective efficacy, thus immune modulators are being considered to enhance the effectiveness of PRRSV vaccines. In this study, the recombinant adenoviruses expressing porcine CD40 ligand (CD40L) and GP3/GP5 of PRRSV were constructed and the immune responses were examined in pigs. The results showed that rAd-CD40L-GP35 (co-expressing CD40L and GP3-GP5) or rAd-GP35 (expressing GP3-GP5) plus rAd-CD40L (expressing CD40L) could provide significant higher specific anti-PRRSV ELISA antibody and neutralizing antibody. And the levels of proliferative responses of peripheral blood mononuclear cells (PBMC), IFN-γ and IL-4 were markedly increased in rAd-CD40L-GP35 and rAd-CD40L plus rAd-GP35 groups than those in rAd-GP35 group. Following homologous challenge with Chinese isolate of the North-American genotype of PRRSV, pigs inoculated with recombinant rAd-CD40L-GP35 and rAd-CD40L plus rAd-GP35 showed lighter clinical signs and lower viremia, as compared to those in rAd-GP35 group. It indicated that porcine CD40L could effectively increase humoral and cell-mediated immune responses of GP3 and GP5 of PRRSV. Porcine CD40L might be used as an attractive adjuvant or immunotargeting strategies to enhance the PRRSV subunit vaccine responses in swine.
Vaccine 11/2010; 28(47):7514-22. · 3.77 Impact Factor
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ABSTRACT: Porcine circovirus type 2 (PCV2) is the essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In the present study, we obtained sequences of 31 PCV2 isolates from different farms of 11 provinces of eastern China and analyzed the genetic characterization of 136 eastern China-derivate PCV2 isolated during 2001-2009. The results showed that these PCV2 isolates could be divided into two groups, PCV2b (108 of 1A/1B, 19 of 1C) and PCV2a (1 of 2A, 2 of 2D, 6 of 2E). Among the 9 PCV2a isolates, eight were found before the year 2005. Meanwhile, three major heterogenic regions were observed in amino acid positions 53-91, 121-151, and 190-210; a few specific substitution patterns were found in each subgroup and several variant or conserved epitopes were also observed in the Cap protein. It indicated that PCV2b predominated in PMWS occurrences in eastern China and a few recombinants of different genotypes existed in nature.
Virus Genes 04/2010; 40(2):244-51. · 1.85 Impact Factor
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ABSTRACT: In ovo vaccination remains an attractive option for a cost effective, uniform and mass application of vaccines for commercial poultry. However, the vaccines which can be delivered safely by this method are limited and there is no currently licensed embryo-safe vaccine against infectious bronchitis virus (IBV). In this study, a recombinant adenovirus expressing the S1 gene of nephropathogenic IBV (rAd-S1) was constructed and the immune responses and protective efficacy against homologous challenge were evaluated after in ovo vaccination. The results showed that the rAd-S1 led to dramatic augmentation of humoral and cellular responses in birds vaccinated in ovo followed by an intramuscular inoculation. Both IFN-gamma and IL-4 in chicken's lymphocytes were produced by this strategy. Following challenge with IBV, the chickens vaccinated with recombinant adenovirus showed fewer nephropathic lesions and less severe clinical signs as compared to those receiving wild-type adenovirus or PBS. The construction of non-replicating human adenovirus vector encoding S1 gene of IBV and its in ovo delivery demonstrated the potential of an alternative vaccination strategy against IBV.
Journal of virological methods 02/2010; 166(1-2):54-9. · 2.13 Impact Factor
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ABSTRACT: Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of postweaning multisystemic wasting syndrome (PMWS). Co-infection of PCV2 and porcine reproductive and respiratory syndrome virus (PRRSV) can result in severe economic losses to the swine industry. In this study, we constructed the recombinant adenovirus rAd-Cap-GP5 expressing Cap of PCV2 and GP5 of PRRSV. And the expression of Cap and GP5 protein in the HEK-293 cells inoculated with rAd-Cap-GP5 were confirmed by immunoperoxidase monolayer assay (IPMA), indirect immunofluorescence assay (IFA) and Western blotting, respectively. The immunogenicity of recombinant adenoviruses rAd-Cap-GP5 was examined in mice by vaccination with the recombinant adenovirus. The results showed that the mice could produce anti-PCV2 and PRRSV antibodies detected by indirect ELISA and virus neutralization assay. It indicated that rAd-Cap-GP5 could provide humoral immunity responses in mice. The recombinant adenovirus rAd-Cap-GP5 might be an attractive candidate vaccine for preventing the disease associated with PCV2 and PRRSV infection.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 11/2009; 25(11):1639-45.
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ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) has recently caused catastrophic losses in swine industry worldwide. Current vaccination strategies only provide a limited protection against PRRSV infection. This study was aimed to construct the recombinant adenovirus co-expressing GP3 and GP5 of highly pathogenic PRRSV fused with swine granulocyte-macrophage colony stimulating factor (GM-CSF) (rAd-GF35), and to detect the immune response in mice and pigs. The results showed that the rAd-GF35 could induce significantly higher PRRSV-specific neutralizing antibodies than the recombinant adenovirus only expressing GP3 and GP5 (rAd-GP35). Moreover, the fusion of GM-CSF markedly increased the secretion of IFN-gamma and IL-4 in PRRSV-stimulated mice lymphocytes culture and pigs sera. Following challenge with PRRSV, piglets inoculated with recombinant rAd-GF35 had lighter clinical signs, lower viremia and less gross lesion of lungs, as compared to that of rAd-GP35 immunized group. It demonstrated that GM-CSF fused with GP3 and GP5 of PRRSV could significantly enhance the humoral and cellular immune responses and provide protection against PRRSV challenge in pigs. The recombinant adenovirus rAd-GF35 might be an attractive candidate vaccine for the prevention and control of highly pathogenic PRRSV infection.
Virus Research 08/2009; 143(1):24-32. · 2.94 Impact Factor
