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1.
Arch Virol ; 164(12): 3059-3063, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31549301

RESUMO

Swine are the only known hosts of swinepox virus (SWPV), the sole member of the genus Suipoxvirus, family Poxviridae. Rapid diagnosis is recommended for appropriate interventions because of the high morbidity associated with this virus. This study describes a real-time quantitative PCR (qPCR) assay for rapid detection and quantification of SWPV. The detection limit, repeatability, reproducibility, and specificity of this assay were determined. The efficiency was 96%, and the R2 value was 0.996. The detection limit was 1 fg or 10-0.5 TCID50/50 µL. Tests showed that the greatest source of error in the SWPV qPCR assay was variation between analysts rather than different qPCR kits or equipment. All nucleic acids from other viruses or samples collected from swine were negative in the specificity test. qPCR for SWPV is a new method with tested variables that allows main sources of error in laboratory diagnosis and viral quantification to be identified.


Assuntos
Infecções por Poxviridae/diagnóstico , Suipoxvirus/genética , Doenças dos Suínos/virologia , Animais , DNA Viral/genética , Limite de Detecção , Infecções por Poxviridae/veterinária , Reação em Cadeia da Polimerase em Tempo Real , Suipoxvirus/classificação , Suipoxvirus/isolamento & purificação , Suínos
2.
Vet Res ; 49(1): 14, 2018 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-29415767

RESUMO

Swinepox virus (SPV) has several advantages as a potential clinical vector for a live vector vaccine. In this study, to obtain a safer and more efficient SPV vector, three SPV mutants, Δ003, Δ010, and ΔTK were successfully constructed. A virus replication experiment showed that these SPV mutants had lower replication abilities compared to wtSPV in 10 different host-derived cell lines. Animal experiments with mouse and rabbit models demonstrate that these three mutants and wtSPV did not cause any clinical signs of dermatitis. No fatalities were observed during a peritoneal challenge assay with these mutants and wtSPV in a mouse model. Additionally, the three mutants and wtSPV were not infectious at 60 h after vaccination in rabbit models. Furthermore, we evaluated biosafety, immunogenicity and effectiveness of the three mutants in 65 1-month-old piglets. The results show that there were no clinical signs of dermatitis in the Δ003 and ΔTK vaccination groups. However, mild signs were observed in the Δ010 vaccination groups when virus titres were high, and apparent clinical signs were observed at the sites of inoculation. Samples from all experimental pig groups were assessed by qPCR, and no SPV genomic DNA was found in five organs, faeces or blood. This suggests that the infectious abilities of wtSPV and the SPV mutants were poor and limited. In summary, this study indicates that two mutants of SPV, Δ003 and ΔTK, may be promising candidates for an attenuated viral vector in veterinary medicine.


Assuntos
Suipoxvirus/genética , Doenças dos Suínos/prevenção & controle , Vacinas Virais/administração & dosagem , Vacinas Virais/imunologia , Animais , Contenção de Riscos Biológicos , Vetores Genéticos/genética , Camundongos , Mutação , Coelhos , Suínos , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia , Vacinas Virais/genética , Replicação Viral
3.
Arch Virol ; 162(12): 3779-3789, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28916870

RESUMO

Porcine epidemic diarrhea virus (PEDV) causes significant loss to the swine industry. The emergence of novel PEDV strains in recent years has decreased the effectiveness of PEDV vaccines. We have developed a live recombinant vaccine, a swinepox virus vector that expresses a truncated S protein (rSPV-St) from a recent PEDV strain, SQ2014, and evaluated its immunogenicity and effectiveness in a swine model. Vaccination of swine with rSPV-St elicited a robust antibody response specific for the homologous PEDV SQ2014. Serum IgA titers in rSPV-St-vaccinated animals were significantly higher than in those immunized with inactivated vaccines. The effectiveness of antibodies induced by the rSPV-St vaccine in protection against PEDV was tested in a passive-transfer model in which piglets were challenged with the homologous virus SQ2014 and the heterologous strain CV777. When challenged with the homologous virus, sera from rSPV-St vaccination provided complete protection. However, sera from rSPV-St vaccination did not provide any protection against the heterologous virus challenge. Amino acid sequence differences in the S proteins of the two viruses were identified within neutralizing epitopes, which might have contributed to the divergent clinical results. Our data suggest that rSPV-St is potentially an effective vaccine against infection with emerging PEDV strains.


Assuntos
Infecções por Coronavirus/veterinária , Portadores de Fármacos , Vírus da Diarreia Epidêmica Suína/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Suipoxvirus/genética , Doenças dos Suínos/prevenção & controle , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Infecções por Coronavirus/prevenção & controle , Proteção Cruzada , Imunização Passiva , Imunoglobulina A/sangue , Vírus da Diarreia Epidêmica Suína/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Glicoproteína da Espícula de Coronavírus/genética , Suínos , Resultado do Tratamento , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/genética
4.
Sci Rep ; 7: 43990, 2017 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-28272485

RESUMO

To characterize neutralizing mimotopes, phages were selected from a 12-mer phage display library using three anti-porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing monoclonal antibodies: (1) A1; (2) A2; and (3) A7. Of these, A2 and A7 recognize the mimotope, P2, which contains the SRHDHIH motif, which has conserved consensus sequences from amino acid positions 156 to 161 in the N-terminal ectodomain of GP3. The artificial multi-epitope gene, mp2, was designed by combining three repeats of the mimotope P2. The resulting sequence was inserted into the swinepox virus (SPV) genome to construct a recombinant swinepox virus (rSPV-mp2). The rSPV-mp2 was able to stably express the multi-epitope peptide, mP2, in vitro. The rSPV-mp2 immunized pigs exhibited a significantly shorter fever duration compared with the wtSPV treated group (P < 0.05). There was an enhanced humoral and cellular immune response, decreased number of PRRSV genomic copies, and a significant reduction in the gross lung pathology (P < 0.05) was observed following PRRSV infection in rSPV-mp2-immunized animals. The results suggest that the recombinant rSPV-mp2 provided pigs with significant protection against PRRSV infection.


Assuntos
Epitopos/genética , Peptídeos/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , Suipoxvirus/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Temperatura Corporal , Epitopos/química , Epitopos/imunologia , Interferon gama/sangue , Interleucina-4/sangue , Pulmão/patologia , Biblioteca de Peptídeos , Peptídeos/química , Peptídeos/imunologia , Síndrome Respiratória e Reprodutiva Suína/metabolismo , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , RNA Viral/sangue , Suipoxvirus/genética , Suipoxvirus/metabolismo , Suínos , Vacinas Virais/genética , Vacinas Virais/imunologia , Vacinas Virais/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-27260812

RESUMO

Swinepox virus (SWPV), a member of the genus Suipoxvirus causes generalized pock-like lesions on the body of domestic and wild pigs. Although outbreak has been reported in India since 1987, virus isolation and genetic characterization remained elusive. In September 2013, an outbreak of acute skin infection occurred in piglets in a commercial piggery unit at Rohtak district in Haryana, India. The presence of SWPV in scab samples collected from piglets succumbed to infection was confirmed by virus isolation, PCR amplification of SWPV-specific gene segments and nucleotide sequencing. Phylogenetic analysis of host-range genes of the SWPV revealed that the Indian isolate is genetically closely related to reference isolate SWPV/pig/U.S.A/1999/Nebraska. To the best of our knowledge this is the first report on isolation and genetic characterization of SWPV from pigs in India.


Assuntos
Especificidade de Hospedeiro , Infecções por Poxviridae/veterinária , Suipoxvirus/genética , Suipoxvirus/isolamento & purificação , Doenças dos Suínos/virologia , Animais , Repetição de Anquirina/genética , Sequência de Bases , Surtos de Doenças , Índia/epidemiologia , Filogenia , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/virologia , Suínos/virologia
6.
J Microbiol Biotechnol ; 26(7): 1173-81, 2016 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-27012234

RESUMO

Salmonella spp. are gram-negative flagellated bacteria that cause a variety of diseases in humans and animals, ranging from mild gastroenteritis to severe systemic infection. To explore development of a potent vaccine against Salmonella infections, the gene encoding outer membrane protein L (ompL) was inserted into the swinepox virus (SPV) genome by homologous recombination. PCR, western blot, and immunofluorescence assays were used to verify the recombinant swinepox virus rSPV-OmpL. The immune responses and protection efficacy of rSPV-OmpL were assessed in a mouse model. Forty mice were assigned to four groups, which were immunized with rSPV-OmpL, inactive Salmonella (positive control), wildtype SPV (wtSPV; negative control), or PBS (challenge control), respectively. The OmpLspecific antibody in the rSPV-OmpL-immunized group increased dramatically and continuously over time post-vaccination, and was present at a significantly higher level than in the positive control group (p < 0.05). The concentrations of IFN-γ and IL-4, which represent Th1-type and Th2-type cytokine responses, were significantly higher (p < 0.05) in the rSPVOmpL- vaccinated group than in the other three groups. After intraperitoneal challenge with a lethal dose of Salmonella typhimurium CVCC542, eight out of ten mice in the rSPV-OmpLvaccinated group were protected, whereas all the mice in the negative control and challenge control groups died within 3 days. Passive immune protection assays showed that hyperimmune sera against OmpL could provide mice with effective protection against challenge from S. typhimurium. The recombinant swinepox virus rSPV-OmpL might serve as a promising vaccine against Salmonella infection.


Assuntos
Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Vetores Genéticos/genética , Salmonella/genética , Salmonella/imunologia , Suipoxvirus/genética , Animais , Citocinas/sangue , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica , Ordem dos Genes , Camundongos , Filogenia , Salmonella/classificação , Infecções por Salmonella/imunologia , Infecções por Salmonella/microbiologia , Infecções por Salmonella/mortalidade , Suínos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo
7.
Methods Mol Biol ; 1349: 163-75, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26458836

RESUMO

Poxvirus-vectors have been widely used in vaccine development for several important human and animal diseases; some of these vaccines have been licensed and used extensively. Swinepox virus (SPV) is well suited to develop recombinant vaccines because of its large packaging capacity for recombinant DNA, its host range specificity, and its ability to induce appropriate immune responses.


Assuntos
Suipoxvirus/genética , Vacinação/métodos , Vacinas Sintéticas/genética , Animais , DNA Recombinante/genética , DNA Recombinante/imunologia , Vetores Genéticos , Humanos , Suipoxvirus/imunologia , Vacinação/veterinária , Vacinas Sintéticas/biossíntese
8.
Vaccine ; 33(32): 3900-6, 2015 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-26116254

RESUMO

To explore the possibility of developing a vaccine against transmissible gastroenteritis virus (TGEV) infection, a recombinant swinepox virus (rSPV-SA) expressing a TGEV protective antigen has been constructed. Immune responses and protection efficacy of the vaccination vector were assessed in both mice and pig models. An indirect ELISA assay suggested that when mice were vaccinated with rSPV-SA, the level of IgG against TGEV was enhanced dramatically. The cytokine assays were employed and the results indicated that both the Th1-type and Th2-type cytokine levels raised after vaccination with rSPV-SA in mice models. Results from the passive immunity protection test of new born piglets demonstrated that the recombinant live-vector vaccine, rSPV-SA, could 100% protect piglets from the SPV infection, and there was no significant clinical symptom in the rSPV-SA treatment group during this experiment. The data suggest that the novel recombinant swinepox virus is a potential vaccine against TGEV infection.


Assuntos
Portadores de Fármacos , Epitopos/imunologia , Gastroenterite Suína Transmissível/prevenção & controle , Suipoxvirus/genética , Vírus da Gastroenterite Transmissível/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Citocinas/metabolismo , Epitopos/genética , Gastroenterite Suína Transmissível/imunologia , Vetores Genéticos , Imunização Passiva , Imunoglobulina G/sangue , Camundongos Endogâmicos BALB C , Suínos , Linfócitos T/imunologia , Vírus da Gastroenterite Transmissível/genética , Resultado do Tratamento , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/genética
9.
Vet Microbiol ; 171(1-2): 198-205, 2014 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-24726504

RESUMO

To develop a vaccine against Porcine circovirus type 2 (PCV2) and Streptococcus equi ssp. zooepidemicus (SEZ) co-infection, the genes of porcine IL-18, capsid protein (Cap) of PCV2 and M-like protein (SzP) of SEZ were inserted into the swinepox virus (SPV) genome by homologous recombination. The recombinant swinepox virus rSPV-ICS was verified by PCR and indirect immunofluorescence assays. To evaluate the immunogenicity of rSPV-ICS, 28 PCV2 and SEZ seronegative Bama minipigs were immunized with rSPV-ICS (n=8), commercial PCV2 vaccine and SEZ vaccine (n=8) or wild type SPV (n=8). The results showed that SzP-specific antibody and PCV2 neutralizing antibody of the rSPV-ICS immunized group increased significantly compared to the wild type SPV treated group after vaccination and increased continuously over time. The levels of IL-4 and IFN-γ in the rSPV-ICS immunized group were significantly higher than the other three groups, respectively. After been co-challenged with PCV2 and SEZ, 87.5% piglets in rSPV-ICS immunized group were survived. Significant reductions in gross lung lesion score, histopathological lung lesion score, and lymph node lesion score were noticed in the rSPV-ICS immunized group compared with the wtSPV treated group. The results suggested that the recombinant rSPV-ICS provided piglets with significant protection against PCV2-SEZ co-infection; thus, it offers proof-of-principle for the development of a vaccine for the prevention of these swine diseases.


Assuntos
Vacinas Bacterianas , Infecções por Circoviridae/veterinária , Imunização/veterinária , Infecções Estreptocócicas/veterinária , Doenças dos Suínos/prevenção & controle , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/normas , Infecções por Circoviridae/complicações , Infecções por Circoviridae/patologia , Infecções por Circoviridae/prevenção & controle , Circovirus/genética , Circovirus/imunologia , Coinfecção , Infecções Estreptocócicas/complicações , Infecções Estreptocócicas/patologia , Infecções Estreptocócicas/prevenção & controle , Streptococcus equi/genética , Streptococcus equi/imunologia , Suipoxvirus/genética , Suínos , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas Virais/genética , Vacinas Virais/imunologia , Vacinas Virais/normas
10.
Arch Virol ; 158(3): 629-37, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23135159

RESUMO

Swine influenza (SI) is an acute respiratory infectious disease of swine caused by swine influenza virus (SIV). SIV is not only an important respiratory pathogen in pigs but also a potent threat to human health. Here, we report the construction of a recombinant swinepox virus (rSPV/H3-2A-H1) co-expressing hemagglutinin (HA1) of SIV subtypes H1N1 and H3N2. Immune responses and protection efficacy of the rSPV/H3-2A-H1 were evaluated in guinea pigs. Inoculation of rSPV/H3-2A-H1 yielded neutralizing antibodies against SIV H1N1 and H3N2. The IFN-γ and IL-4 concentrations in the supernatant of lymphocytes stimulated with purified SIV HA1 antigen were significantly higher (P < 0.01) than those of the control groups. Complete protection of guinea pigs against SIV H1N1 or H3N2 challenge was observed. No SIV shedding was detected from guinea pigs vaccinated with rSPV/H3-2A-H1 after challenge. Most importantly, the guinea pigs immunized with rSPV/H3-2A-H1 did not show gross and micrographic lung lesions. However, the control guinea pigs experienced distinct gross and micrographic lung lesions at 7 days post-challenge. Our data suggest that the recombinant swinepox virus encoding HA1 of SIV H1N1 and H3N2 might serve as a promising candidate vaccine for protection against SIV H1N1 and H3N2 infections.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza , Infecções por Orthomyxoviridae/veterinária , Suipoxvirus/genética , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Linhagem Celular , Citocinas/biossíntese , Citocinas/imunologia , Cães , Cobaias , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/genética , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/virologia , Suipoxvirus/imunologia , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/prevenção & controle , Células Th1/imunologia , Células Th2/imunologia , Vacinação/veterinária , Vacinas Sintéticas/imunologia
11.
Vet Microbiol ; 162(1): 259-64, 2013 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-23265244

RESUMO

The recombinant swine poxvirus rSPV/H3-2A-H1 co-expressing HA1 genes of H3N2 and H1N1 subtype SIV has been constructed and identified. Inoculations of rSPV/H3-2A-H1 yielded ELISA and neutralization antibodies against SIV H1N1 and H3N2, and elicited potent H1N1 and H3N2 SIV-specific INF-γ response from T-lymphocytes in mice and pigs in this study. Complete protection against SIV H1N1 or H3N2 challenge in pigs was observed.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/genética , Vacinas contra Influenza/genética , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/veterinária , Suipoxvirus/genética , Doenças dos Suínos/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza/farmacologia , Interferon gama/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/virologia , Distribuição Aleatória , Suipoxvirus/imunologia , Sus scrofa , Suínos , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/virologia , Linfócitos T/imunologia , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/farmacologia
12.
Vaccine ; 30(44): 6307-13, 2012 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-22884664

RESUMO

To explore development of a vaccine against PCV2 infections, the gene of capsid protein (Cap) was inserted into the swinepox virus (SPV) genome by homologous recombination. The recombinant swinepox virus expressing capsid protein (rSPV-cap) was verified by PCR, western blot and immunofluorescence assays. To evaluate the immunogenicity of rSPV-cap, twenty-four PCV2 seronegative minipigs were immunized with rSPV-cap, wild type SPV (wtSPV; negative control), or PBS (challenge control). After inoculation with PCV2, pigs in the rSPV-cap immunized group showed significantly higher average daily weight gain (ADG) and shorter fever duration compared with the wtSPV treated group (P<0.05). Cap-specific antibody in the rSPV-cap immunized group increased dramatically after vaccination and increased continuously over time. PCV2 genomic copies in the serum of rSPV-cap immunized pigs were significantly less compared with the wtSPV treated group at all time points after inoculation (P<0.01). Significant reduction in gross lung lesion scores, histopathological lung lesion scores, and lymph node lesion scores were noted in the rSPV-cap immunized group compared with the wtSPV treated group (P<0.01). The results suggested that the recombinant rSPV-cap provided pigs with significant protection from PCV2-associated disease; thus, it offers proof-of-principle for the development of a vaccine for the prevention of PCV2-associated disease in pigs.


Assuntos
Proteínas do Capsídeo/imunologia , Infecções por Circoviridae/veterinária , Circovirus/imunologia , Suipoxvirus/genética , Doenças dos Suínos/prevenção & controle , Vacinas Virais/administração & dosagem , Vacinas Virais/imunologia , Animais , Peso Corporal , Proteínas do Capsídeo/genética , Infecções por Circoviridae/patologia , Infecções por Circoviridae/prevenção & controle , Circovirus/genética , Portadores de Fármacos , Febre/prevenção & controle , Vetores Genéticos , Histocitoquímica , Pulmão/patologia , Linfonodos/patologia , Suínos , Doenças dos Suínos/patologia , Porco Miniatura , Fatores de Tempo , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Virais/genética
13.
Virus Res ; 167(2): 188-95, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22584406

RESUMO

Swine influenza virus (SIV) is not only an important respiratory pathogen in pigs but also a potent threat to human health. Even though immunization with recombinant vaccinia poxviruses expressing protective antigens as a vaccination strategy has been widely used for many infectious diseases, development of recombinant swinepox virus (rSPV) vector for this purpose has been less successful. Here, we report the construction of a recombinant swinepox virus (rSPV) expressing hemagglutinin (HA1) of H3N2 SIV (rSPV-H3). Immune responses and protection efficacy of the vaccination vector were assessed in both mouse and pig models. Prime and boost inoculations of rSPV-H3 yielded neutralization antibody against SIV and elicited potent H3N2 SIV-specific INF-γ response from T-lymphocytes. Complete protection of pigs against H3N2 SIV challenge was achieved. No pigs showed severe systemic and local reactions and no SIV was found shed from the pigs vaccinated with rSPV-H3 after challenge. The data suggest that the SPV-based recombinant vector expressing HA1 of H3N2 SIV might serve as a promising SIV vaccine for protection against SIV infection.


Assuntos
Portadores de Fármacos/administração & dosagem , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Suipoxvirus/genética , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Feminino , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H3N2/genética , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Interferon gama/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/patologia , Infecções por Orthomyxoviridae/virologia , Suínos , Linfócitos T/imunologia , Vacinação/métodos , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Eliminação de Partículas Virais
14.
Berl Munch Tierarztl Wochenschr ; 125(3-4): 144-52, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22515033

RESUMO

To explore the potential of the swinepox virus (SPV) as vector for Streptococcus suis vaccines, a vector system was developed for the construction of a recombinant SPV carrying bacterial genes. Using this system, a recombinant virus expressing truncated muramidase-released protein (MRP) of S. suis type 2 (SS2), designated rSPV-MRP, was produced and identified by PCR, western blotting and immunofluorescence assays. The rSPV-MRP was found to be only slightly attenuated in PK-15 cells, when compared with the wild-type virus. After immunization intramuscularly with rSPV-MRP, SS2 inactive vaccine (positive control), wild-type SPV (negative control) and PBS (blank control) respectively, all CD1 mice were challenged with a lethal dose or a sublethal dose of SS2 highly virulent strain ZY05719. While SS2 inactive vaccine protected all mice, immunization with rSPV-MRP resulted in 60% survival and protected mice against a lethal dose of the highly virulent SS2 strain, compared with the negative control (P < 0.05). Our data indicate that animals immunized with rSPV-MRP had a significantly reduced bacterial burden in all organs examined, compared to negative controls and blank controls (P <0.05). Antibody titers of the rSPV-MRP-vaccinated group were significantly higher (P <0.001), when compared to negative controls and blank controls. Antibody titers were also significantly higher in the vaccinated group at all time points post-vaccination (P <0.001), compared with the positive controls. These initial results demonstrated that the rSPV-MRP provided mice with protection from systemic SS2 infection. If SPV recombinants have the potential as S. suis vaccines for the use in pigs has to be evaluated in further studies.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas , Infecções Estreptocócicas/veterinária , Streptococcus suis/imunologia , Suipoxvirus/genética , Doenças dos Suínos/prevenção & controle , Animais , Anticorpos Antibacterianos/biossíntese , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Vacinas Bacterianas/genética , Vacinas Bacterianas/imunologia , Linhagem Celular , Feminino , Regulação Bacteriana da Expressão Gênica , Vetores Genéticos , Camundongos , Reação em Cadeia da Polimerase/veterinária , Distribuição Aleatória , Vírus Reordenados/genética , Vírus Reordenados/imunologia , Inoculações Seriadas , Infecções Estreptocócicas/prevenção & controle , Streptococcus suis/genética , Suipoxvirus/imunologia , Suínos , Vacinação/veterinária , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
15.
Vaccine ; 30(20): 3119-25, 2012 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-22391400

RESUMO

Swine influenza virus (SIV) is not only an important respiratory pathogen in pigs but also a potent threat to human health. Although immunization with recombinant poxviruses expressing protective antigens as vaccines has been widely used for against many infectious diseases, development of recombinant swinepox virus (rSPV) vector for the purpose has been less successful. Here, we report the construction of a recombinant swinepox virus (rSPV-HA1) expressing hemagglutinin (HA1) of H1N1 SIV. Immune responses and protection efficacy of the vaccination vector were evaluated in both the mouse model and the natural host: pig. Prime and boost inoculations of rSPV-HA1 yielded high levels of neutralization antibody against SIV and elicited potent H1N1 SIV-specific IFN-γ response from T-lymphocytes. Complete protection of pigs against H1N1 SIV challenge was observed. No pigs showed evident systemic and local reactions to the vaccine and no SIV shedding was detected from pigs vaccinated with rSPV-HA1 after challenge. Our data demonstrated that the recombinant swinepox virus encoding HA1 of SIV H1N1 may serve as a promising SIV vaccine for protection against SIV infection.


Assuntos
Vetores Genéticos , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vacinas contra Influenza/imunologia , Suipoxvirus/genética , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Modelos Animais de Doenças , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Imunização Secundária/métodos , Vírus da Influenza A Subtipo H1N1/genética , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Interferon gama/metabolismo , Camundongos , Infecções por Orthomyxoviridae/prevenção & controle , Suínos , Linfócitos T/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
16.
J Virol ; 86(8): 4538-47, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22345458

RESUMO

Vaccinia virus (VACV) K1L and C7L function equivalently in many mammalian cells to support VACV replication and antagonize antiviral activities induced by type I interferons (IFNs). While K1L is limited to orthopoxviruses, genes that are homologous to C7L are found in diverse mammalian poxviruses. In this study, we showed that the C7L homologues from sheeppox virus and swinepox virus could rescue the replication defect of a VACV mutant deleted of both K1L and C7L (vK1L(-)C7L(-)). Interestingly, the sheeppox virus C7L homologue could rescue the replication of vK1L(-)C7L(-) in human HeLa cells but not in murine 3T3 and LA-4 cells, in contrast to all other C7L homologues. Replacing amino acids 134 and 135 of the sheeppox virus C7L homologue, however, made it functional in the two murine cell lines, suggesting that these two residues are critical for antagonizing a putative host restriction factor which has some subtle sequence variation in human and murine cells. Furthermore, the C7L family of host range genes from diverse mammalian poxviruses were all capable of antagonizing type I IFN-induced antiviral activities against VACV. Screening of a library of more than 350 IFN-stimulated genes (ISGs) identified interferon-regulated factor 1 (IRF1) as an inhibitor of vK1L(-)C7L(-) but not wild-type VACV. Expression of either K1L or C7L, however, rendered vK1L(-)C7L(-) resistant to IRF1-induced antiviral activities. Altogether, our data show that K1L and C7L antagonize IRF1-induced antiviral activities and that the host modulation function of C7L is evolutionally conserved in all poxviruses that can readily replicate in tissue-cultured mammalian cells.


Assuntos
Especificidade de Hospedeiro/genética , Fator Regulador 1 de Interferon/metabolismo , Interferon Tipo I/metabolismo , Poxviridae/genética , Poxviridae/metabolismo , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Capripoxvirus/genética , Capripoxvirus/metabolismo , Linhagem Celular , Chlorocebus aethiops , Ordem dos Genes , Humanos , Fator Regulador 1 de Interferon/genética , Camundongos , Dados de Sequência Molecular , Alinhamento de Sequência , Suipoxvirus/genética , Suipoxvirus/metabolismo , Vírus Vaccinia/genética , Vírus Vaccinia/metabolismo , Proteínas Virais/metabolismo , Replicação Viral/genética
18.
Vaccine ; 29(40): 7027-34, 2011 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-21807055

RESUMO

To develop a safer, more immunogenic and efficacious vaccine against Streptococcus equi ssp. zooepidemicus (SEZ) infections, the gene of M-like protein (SzP) was placed under the strong vaccinia virus promoter P28 and then inserted into swinepox virus (SPV) genome. The recombinant swinepox virus (rSPV-szp) was isolated in a non-selective medium by the co-expression of Escherichia coli LacZ gene and verified by PCR, western blotting and immunofluorescence assays. To evaluate the immunogenicity of this rSPV-szp, ICR mice were immunized with the rSPV-szp, inactivated SEZ vaccine (positive control), wild type SPV (negative control), or PBS (challenge control). All mice were intraperitoneally challenged with 5 LD(50) of homogenous ATCC 35246 strain 14 days post-vaccination. The results showed that at least 70% mice in rSPV-szp-vaccinated group were protected against homogenous ATCC 35246 challenge, the survival rate was significantly higher compared with mice in the negative control group and the challenge control group (P<0.001). The antibody titers of the rSPV-szp-vaccinated group were significantly higher (P<0.05) than the other three groups. Passive immune protection assays showed that the hyperimmune sera against M-like protein could provide mice with complete protection against challenge of ATCC 35246. Semi-quantitative RT-PCR analysis showed a marked increased in levels of IL-4 and IFN-γ mRNA in immunized mice. The results suggested that the recombinant rSPV-szp provided mice with significant protection from the SEZ infections. It is a promising candidate for the vaccine development against SEZ infections.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas de Transporte/imunologia , Vacinas Antiestafilocócicas/genética , Vacinas Antiestafilocócicas/imunologia , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/prevenção & controle , Streptococcus equi/imunologia , Suipoxvirus/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Células Cultivadas , Escherichia coli/genética , Feminino , Imunidade Celular/imunologia , Imunização Passiva/métodos , Interferon gama/genética , Interferon gama/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Camundongos , Camundongos Endogâmicos ICR , Plasmídeos/genética , Regiões Promotoras Genéticas , Vacinas Antiestafilocócicas/farmacologia , Suipoxvirus/genética , Suínos , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/farmacologia , Vírus Vaccinia/genética , Vírus Vaccinia/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia
19.
Dtsch Tierarztl Wochenschr ; 115(4): 162-6, 2008 Apr.
Artigo em Alemão | MEDLINE | ID: mdl-18500151

RESUMO

Swinepox virus infection results in an acute, mild or subclinical course and is characterised by typical poxvirus skin lesions in affected pigs. Additionally, sporadic vertical swinepox virus transmission leads to congenital generalised infection and subsequent abortion or stillbirth. The present report describes the occurrence of epidermal efflorescences in two piglets after intrauterine natural suipoxvirus infection. No clinical abnormalities of the gilt and littermates as well as in other pigs from this herd were present. One of the affected piglets was stillborn and submitted for necropsy, the other animal was alive at birth, but died 3 days later. Histologically, a proliferative to ulcerative dermatitis with epithelial ballooning degeneration and characteristic intracytoplasmatic inclusion bodies was observed. The pathomorphological and histopathological suspected diagnosis of a poxvirus infection was confirmed by electron microscopy. Furthermore, the agent was identified as suipoxvirus by polymerase chain reaction. As demonstrated here, obvious skin lesions in suipoxvirus infection leads to a suspected diagnosis in newborn piglets on macroscopic examination. However, further post mortem examinations, including electron microscopy as well as molecular techniques are essential for the identification of the aetiology and the exclusion of differential diagnoses. Because the disease only affected two pigs there was only a small economic loss. A valid diagnostic plays an important role in advising farmers and for herd health monitoring.


Assuntos
Transmissão Vertical de Doença Infecciosa/veterinária , Infecções por Poxviridae/veterinária , Complicações Infecciosas na Gravidez/veterinária , Suipoxvirus , Doenças dos Suínos/transmissão , Animais , Animais Recém-Nascidos , Evolução Fatal , Feminino , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/patologia , Infecções por Poxviridae/transmissão , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Complicações Infecciosas na Gravidez/patologia , Pele/patologia , Pele/virologia , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/patologia
20.
Vet Microbiol ; 111(1-2): 1-13, 2005 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-16181751

RESUMO

Costimulatory ligands, B7.1 and B7.2, have been incorporated into viral and DNA vectors as potential nonchemical adjuvants to enhance CTL and humoral immune responses against viral pathogens. In addition, soluble B7 proteins, minus their transmembrane and cytoplasmic domains, have been shown to block the down regulation of T-cell activation through blockade of B7/CTLA-4 interactions in mouse tumor models. Recently, we developed swinepox virus (SPV) vectors for delivery of feline leukemia antigens for vaccine use in cats [Winslow, B.J., Cochran, M.D., Holzenburg, A., Sun, J., Junker, D.E., Collisson, E.W., 2003. Replication and expression of a swinepox virus vector delivering feline leukemia virus Gag and Env to cell lines of swine and feline origin. Virus Res. 98, 1-15]. To explore the use of feline B7.1 and B7.2 ligands as nonchemical adjuvants, SPV vectors containing full-length feline B7.1 and B7.2 ligands were constructed and analyzed. Full-length feline B7.1 and B7.2 produced from SPV vectors were natively processed and costimulated Jurkat cells to produce IL-2, in vitro. In addition, we explored the feasibility of utilizing SPV as a novel expression vector to produce soluble forms of feline B7.1 (sB7.1) and B7.2 (sB7.2) in tissue culture. The transmembrane and cytoplasmic regions of the B7.1 and B7.2 genes were replaced with a poly-histidine tag and purified via a two-step chromatography procedure. Receptor binding and costimulation activity was measured. Although feline sB7.1-his and sB7.2-his proteins bound to the human homolog receptors, CTLA-4 and CD28, both soluble ligands possessed greater affinity for CTLA-4, compared to CD28. However, both retained the ability to partially block CD28-mediated costimulation in vitro. Results from these studies establish the use of SPV as a mammalian expression vector and suggest that full-length-vectored and purified soluble feline B7 ligands may be valuable, nonchemical immune-modulators.


Assuntos
Adjuvantes Imunológicos , Antígeno B7-1/imunologia , Antígeno B7-2/imunologia , Infecções por Poxviridae/veterinária , Suipoxvirus/imunologia , Animais , Antígenos CD , Antígenos de Diferenciação , Antígeno B7-1/genética , Antígeno B7-2/genética , Antígeno CTLA-4 , Gatos , Linhagem Celular , Produtos do Gene env/metabolismo , Produtos do Gene gag/metabolismo , Vetores Genéticos , Humanos , Imunoconjugados , Interleucina-2/biossíntese , Células Jurkat , Leucemia Felina/prevenção & controle , Ativação Linfocitária , Infecções por Poxviridae/imunologia , Replicação Viral
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