RESUMEN
In September 2022, deaths of pigs manifesting pox-like lesions caused by swinepox virus were reported in Tshuapa Province, Democratic Republic of the Congo. Two human mpox cases were found concurrently in the surrounding community. Specific diagnostics and robust sequencing are needed to characterize multiple poxviruses and prevent potential poxvirus transmission.
Asunto(s)
Mpox , Poxviridae , Suipoxvirus , Humanos , Animales , Porcinos , Mpox/epidemiología , Monkeypox virus/genética , República Democrática del Congo/epidemiologíaRESUMEN
Swinepox is a sporadic virus disease of domestic and wild pigs that mainly occurs during the rainy season. Though the disease is known for a century, research on swinepox especially genetic characterization is scanty. Self-limiting nature of the disease, the non-availability of specific diagnostics as well as the resemblance of clinical signs with other pathogens are some of the issues in the slow progress in swinepox-related research. Recent whole genome sequencing data from the USA, India, and Germany enhanced our understanding of the biology of swinepox virus (SWPV). The objective of the present study is to investigate the molecular epidemiology of two swinepox outbreaks that occurred in 2015 and 2016 one each in Uttar Pradesh, and the Haryana states of India. The appearance of clinical signs in different swine breeds was recorded. The scab samples from infected pigs were collected, DNA extracted, host range genes of SWPV were PCR amplified, sequenced and analyzed for genetic and phylogenetic characterization. Desi (nondescript breed), Yorkshire White pigs, and Landrace cross were found to be infected with SWPV. Host range genes of SWPV analyzed from clinical samples showed very high nucleotide identity with each other. Phylogenetic analyses revealed that SWPVs circulating in India are distinct (Indian lineage) from the SWPV of the USA, Germany, and Russia (European-North American lineage). Our study affirms the existence of two distinct lineages of SWPV globally with differences in clinical lesions between breeds.
Asunto(s)
Infecciones por Poxviridae , Suipoxvirus , Enfermedades de los Porcinos , Porcinos , Animales , Suipoxvirus/genética , Filogenia , Epidemiología Molecular , Infecciones por Poxviridae/epidemiología , Infecciones por Poxviridae/veterinaria , Infecciones por Poxviridae/diagnóstico , Reacción en Cadena de la Polimerasa/veterinaria , India/epidemiología , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Salmonella spp. poses a great threat to the livestock, food safety and public health. A recombinant swinepox virus expressing a protective antigen sseB was constructed by homologous recombination to develop a vaccine against Salmonella infection. The rSPV-sseB was verified using PCR, Western blot and indirect immunofluorescence assay. The immune responses and protective efficacy of rSPV-sseB were assessed in piglets. Forty piglets were immunized with rSPV-sseB, inactive Salmonella vaccine, wild-type SPV (wtSPV), or PBS. The results showed that the level of the sseB-specific antibody of the rSPV-sseB-vaccinated piglets was significantly higher at all time points post-vaccination than those of the inactivated Salmonella vaccine (P < 0.05), wtSPV (P < 0.001) or mock treated piglets (P < 0.001). The IL-4 and IFN-γ in the rSPV-sseB group were significantly higher than the other three groups at all post-infection time points. rSPV-sseB provided piglets with strong protection against the challenge of S. typhimurium with lethal dose. These results suggest the possibility of using recombinant swinepox virus rSPV-sseB as a promising vaccine to prevent Salmonella infection.
Asunto(s)
Infecciones por Salmonella , Vacunas contra la Salmonella , Suipoxvirus , Animales , Porcinos , Suipoxvirus/genética , Salmonella typhimurium/genética , Interleucina-4 , Vacunas SintéticasRESUMEN
Swinepox virus (SWPV) is a globally distributed swine pathogen that causes sporadic cases of an acute poxvirus infection in domesticated pigs, characterized by the development of a pathognomonic proliferative dermatitis and secondary ulcerations. More severe disease with higher levels of morbidity and mortality is observed in congenitally SWPV-infected neonatal piglets. In this study, we investigated the evolutionary origins of SWPV strains isolated from domestic pigs and wild boar. Analysis of whole genome sequences of SWPV showed that at least two different virus strains are currently circulating in Germany. These were more closely related to a previously characterized North American SWPV strain than to a more recent Indian SWPV strain and showed a variation in the SWPV-specific genome region. A single nucleotide deletion in the wild boar (wb) SWPV strain leads to the fusion of the SPV019 and SPV020 open reading frames (ORFs) and encodes a new hypothetical 113 aa protein (SPVwb020-019). In addition, the domestic pig (dp) SWPV genome contained a novel ORF downstream of SPVdp020, which encodes a new hypothetical 71aa protein (SPVdp020a). In summary, we show that SWPV strains with altered coding capacity in the SWPV specific genome region are circulating in domestic pig and wild boar populations in Germany.
Asunto(s)
Infecciones por Poxviridae/veterinaria , Infecciones por Poxviridae/virología , Suipoxvirus/aislamiento & purificación , Sus scrofa/virología , Enfermedades de los Porcinos/virología , Porcinos/virología , Animales , Evolución Molecular , Alemania , Secuenciación de Nucleótidos de Alto Rendimiento , Sistemas de Lectura Abierta , Filogenia , Poxviridae/clasificación , Poxviridae/genética , Especificidad de la Especie , Suipoxvirus/clasificación , Suipoxvirus/genéticaRESUMEN
In this study, we report the complete genome sequence of swinepox virus from a clinical sample from a naturally occurring infection in India. The sequencing was done on a Nanopore MinION sequencer from Oxford Nanopore Technologies. Two new annotations were added to the genome. Three of the genes were found to have frameshifts, which might be of importance in relation to infection. When compared to the only other reported whole genome sequence of swinepox virus, which was obtained from an isolate from America in 1999, our sequence is only 98.19% identical at the nucleotide level. The average amino acid sequence identity of the viral proteins, based on the common 149 annotations, is also 98.19%, demonstrating that these viruses are distinctly divergent. Owing to the fact that swinepox virus infects only swine, it could not have entered America until the introduction of swine in the 16th century from Europe. The swinepox viruses in both continents have continued to evolve independently. The sequence divergence identified here indicates a Eurasian-lineage virus that is geographically distinct from the American-lineage swinepox virus.
Asunto(s)
Genoma Viral/genética , Infecciones por Poxviridae/veterinaria , Suipoxvirus/genética , Enfermedades de los Porcinos/virología , Animales , ADN Viral/genética , Variación Genética , India , Infecciones por Poxviridae/virología , Análisis de Secuencia de ADN , Homología de Secuencia , Suipoxvirus/clasificación , Suipoxvirus/aislamiento & purificación , Porcinos , Proteínas Virales/genéticaRESUMEN
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.
Asunto(s)
Infecciones por Poxviridae/diagnóstico , Suipoxvirus/genética , Enfermedades de los Porcinos/virología , Animales , ADN Viral/genética , Límite de Detección , Infecciones por Poxviridae/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa , Suipoxvirus/clasificación , Suipoxvirus/aislamiento & purificación , PorcinosRESUMEN
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.
Asunto(s)
Suipoxvirus/genética , Enfermedades de los Porcinos/prevención & control , Vacunas Virales/administración & dosificación , Vacunas Virales/inmunología , Animales , Contención de Riesgos Biológicos , Vectores Genéticos/genética , Ratones , Mutación , Conejos , Porcinos , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Vacunas Virales/genética , Replicación ViralRESUMEN
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.
Asunto(s)
Infecciones por Coronavirus/veterinaria , Portadores de Fármacos , Virus de la Diarrea Epidémica Porcina/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Suipoxvirus/genética , Enfermedades de los Porcinos/prevención & control , Vacunas Virales/inmunología , Animales , Anticuerpos Antivirales/sangre , Infecciones por Coronavirus/prevención & control , Protección Cruzada , Inmunización Pasiva , Inmunoglobulina A/sangre , Virus de la Diarrea Epidémica Porcina/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Glicoproteína de la Espiga del Coronavirus/genética , Porcinos , Resultado del Tratamiento , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Virales/administración & dosificación , Vacunas Virales/genéticaRESUMEN
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.
Asunto(s)
Epítopos/genética , Péptidos/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/metabolismo , Suipoxvirus/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Temperatura Corporal , Epítopos/química , Epítopos/inmunología , Interferón gamma/sangre , Interleucina-4/sangre , Pulmón/patología , Biblioteca de Péptidos , Péptidos/química , Péptidos/inmunología , Síndrome Respiratorio y de la Reproducción Porcina/metabolismo , Síndrome Respiratorio y de la Reproducción Porcina/prevención & control , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/inmunología , ARN Viral/sangre , Suipoxvirus/genética , Suipoxvirus/metabolismo , Porcinos , Vacunas Virales/genética , Vacunas Virales/inmunología , Vacunas Virales/metabolismoRESUMEN
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.
Asunto(s)
Especificidad del Huésped , Infecciones por Poxviridae/veterinaria , Suipoxvirus/genética , Suipoxvirus/aislamiento & purificación , Enfermedades de los Porcinos/virología , Animales , Repetición de Anquirina/genética , Secuencia de Bases , Brotes de Enfermedades , India/epidemiología , Filogenia , Infecciones por Poxviridae/epidemiología , Infecciones por Poxviridae/virología , Porcinos/virologíaRESUMEN
This article describes five outbreaks of swinepox in backyard pigs in Northeastern Brazil. It affected backyard pigs from herds of poor hygienic-sanitary conditions with severe fly and lice infestations. The morbidity ranged from 33.3 to 100% among affected herds, with mortality reaching up to 60%. The affected pigs developed multifocal to coalescent gray to white papules and blisters in the skin, with eventual eruptions, evolving to erosions and crusts. In addition to skin lesions, affected piglets presented apathy, anorexia and fever. The disease was auto-limiting, resolving within 15 to 25 days. Histological examination revealed proliferative and ulcerative vesiculopustular dermatitis with ballooning degeneration of epithelial cells, perivascular inflammatory infiltrates of lymphocytes, plasma cells, neutrophils, eosinophils and some macrophages in the dermis. Intracytoplasmic eosinophilic inclusions were consistently observed in keratinocytes. Total DNA extracted from fresh tissue fragments obtained from one outbreak and formalin-fixed, paraffin-embedded (FFPE) tissue from the other four outbreaks was submitted to polymerase chain reaction (PCR) for Swinepox virus (SWPV) and Vaccinia virus (VACV). Genetic SWPV material was identified by PCR in fresh material from one outbreak. Nucleotide sequencing and phylogenetic analysis of the PCR amplicons (viral polymerase gene) demonstrated 100% homology with sequences from SWPV. All tissues were PCR negative for VACV. Swine poxvirus is present in backyard pigs in Northeastern Brazil, indicating the need of including SWPV in the differential diagnosis of dermatitis in pigs.(AU)
Em cinco surtos de varíola em suínos no Nordeste do Brasil foram acometidos leitões e suínos adultos, de rebanhos domésticos criados em condições higiênico-sanitárias precárias, que apresentavam graves infestações por moscas e piolhos. A morbidade variou de 33,3-100% entre os rebanhos afetados e a mortalidade atingindo 60%. Os animais afetados desenvolveram pápulas cinzentas ou esbranquiçadas coalescentes e vesículas, que evoluíram para erosões e crostras. Além das lesões de pele, os leitões afetados apresentavam apatia, anorexia e febre. A doença foi autolimitante, com resolução em 15 a 25 dias. Histologicamente, observou-se dermatite proliferativa e ulcerativa com degeneração balonosa das células do epitélio, infiltrado inflamatório perivascular de linfócitos, plasmócitos, neutrófilos, eosinófilos e escassos macrófagos na derme. Inclusões eosinofílicas intracitoplasmáticas foram consistentemente observadas em queratinócitos. DNA total extraído a partir de fragmentos de tecido frescos obtidos a partir de um surto, e de tecido fixado em formol e embebido em parafina dos outros quatro surtos, foram submetidos à reação em cadeia da polimerase (PCR) para o vírus da varíola suína (SWPV) e o vírus vaccínia (VACV). Material genético do SWPV foi identificado por PCR em material fresco de um surto. O sequenciamento e análise filogenética dos produtos de amplificação da PCR (gene da polimerase viral) demonstraram 100% de homologia com sequências do SWPV. Todos os fragmentos de tecidos foram negativos para VACV na PCR. Este artigo relata a circulação de poxvírus suíno no Nordeste do Brasil, indicando a necessidade de incluir SWPV no diagnóstico diferencial de dermatite em suínos.(AU)
Asunto(s)
Animales , Viruela/epidemiología , Viruela/etiología , Suipoxvirus/aislamiento & purificación , Porcinos/virología , Dermatitis/veterinaria , Reacción en Cadena de la Polimerasa/veterinariaRESUMEN
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.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/inmunología , Vectores Genéticos/genética , Salmonella/genética , Salmonella/inmunología , Suipoxvirus/genética , Animales , Citocinas/sangre , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Orden Génico , Ratones , Filogenia , Salmonella/clasificación , Infecciones por Salmonella/inmunología , Infecciones por Salmonella/microbiología , Infecciones por Salmonella/mortalidad , Porcinos , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/metabolismoRESUMEN
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.
Asunto(s)
Suipoxvirus/genética , Vacunación/métodos , Vacunas Sintéticas/genética , Animales , ADN Recombinante/genética , ADN Recombinante/inmunología , Vectores Genéticos , Humanos , Suipoxvirus/inmunología , Vacunación/veterinaria , Vacunas Sintéticas/biosíntesisRESUMEN
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.
Asunto(s)
Portadores de Fármacos , Epítopos/inmunología , Gastroenteritis Porcina Transmisible/prevención & control , Suipoxvirus/genética , Virus de la Gastroenteritis Transmisible/inmunología , Vacunas Virales/inmunología , Animales , Anticuerpos Antivirales/sangre , Citocinas/metabolismo , Epítopos/genética , Gastroenteritis Porcina Transmisible/inmunología , Vectores Genéticos , Inmunización Pasiva , Inmunoglobulina G/sangre , Ratones Endogámicos BALB C , Porcinos , Linfocitos T/inmunología , Virus de la Gastroenteritis Transmisible/genética , Resultado del Tratamiento , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Virales/administración & dosificación , Vacunas Virales/genéticaRESUMEN
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.
Asunto(s)
Vacunas Bacterianas , Infecciones por Circoviridae/veterinaria , Inmunización/veterinaria , Infecciones Estreptocócicas/veterinaria , Enfermedades de los Porcinos/prevención & control , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/normas , Infecciones por Circoviridae/complicaciones , Infecciones por Circoviridae/patología , Infecciones por Circoviridae/prevención & control , Circovirus/genética , Circovirus/inmunología , Coinfección , Infecciones Estreptocócicas/complicaciones , Infecciones Estreptocócicas/patología , Infecciones Estreptocócicas/prevención & control , Streptococcus equi/genética , Streptococcus equi/inmunología , Suipoxvirus/genética , Porcinos , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Proteínas Virales/genética , Vacunas Virales/inmunología , Vacunas Virales/normasRESUMEN
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.
Asunto(s)
Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/genética , Vacunas contra la Influenza/genética , Vacunas contra la Influenza/inmunología , Infecciones por Orthomyxoviridae/veterinaria , Suipoxvirus/genética , Enfermedades de los Porcinos/inmunología , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Subtipo H1N1 del Virus de la Influenza A/inmunología , Subtipo H3N2 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/farmacología , Interferón gamma/inmunología , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Infecciones por Orthomyxoviridae/virología , Distribución Aleatoria , Suipoxvirus/inmunología , Sus scrofa , Porcinos , Enfermedades de los Porcinos/prevención & control , Enfermedades de los Porcinos/virología , Linfocitos T/inmunología , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/farmacologíaRESUMEN
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.
Asunto(s)
Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Subtipo H1N1 del Virus de la Influenza A/inmunología , Subtipo H3N2 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza , Infecciones por Orthomyxoviridae/veterinaria , Suipoxvirus/genética , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Línea Celular , Citocinas/biosíntesis , Citocinas/inmunología , Perros , Cobayas , Glicoproteínas Hemaglutininas del Virus de la Influenza/biosíntesis , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/genética , Vacunas contra la Influenza/inmunología , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Infecciones por Orthomyxoviridae/virología , Suipoxvirus/inmunología , Porcinos , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/prevención & control , Células TH1/inmunología , Células Th2/inmunología , Vacunación/veterinaria , Vacunas Sintéticas/inmunologíaRESUMEN
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.
Asunto(s)
Proteínas de la Cápside/inmunología , Infecciones por Circoviridae/veterinaria , Circovirus/inmunología , Suipoxvirus/genética , Enfermedades de los Porcinos/prevención & control , Vacunas Virales/administración & dosificación , Vacunas Virales/inmunología , Animales , Peso Corporal , Proteínas de la Cápside/genética , Infecciones por Circoviridae/patología , Infecciones por Circoviridae/prevención & control , Circovirus/genética , Portadores de Fármacos , Fiebre/prevención & control , Vectores Genéticos , Histocitoquímica , Pulmón/patología , Ganglios Linfáticos/patología , Porcinos , Enfermedades de los Porcinos/patología , Porcinos Enanos , Factores de Tiempo , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Virales/genéticaRESUMEN
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.
Asunto(s)
Portadores de Fármacos/administración & dosificación , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Subtipo H3N2 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Suipoxvirus/genética , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Femenino , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Subtipo H3N2 del Virus de la Influenza A/genética , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/genética , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/patología , Infecciones por Orthomyxoviridae/virología , Porcinos , Linfocitos T/inmunología , Vacunación/métodos , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Esparcimiento de VirusRESUMEN
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.