RESUMO
The Bovine herpes virus type 5 glycoprotein D (gD) is essential for viral penetration into host permissive cells. The Herpes virus gD glycoprotein has been used for bovine immunization, being efficient in reduction of viral replication, shedding and clinical signs, however sterilizing immunity is still not achieved. Recombinant subunit vaccines are, in general, poorly immunogenic requiring additional adjuvant components. Interleukin 17A (IL17A) is a pro-inflammatory cytokine produced by T helper 17 cells that mediate mucosal immunity. IL17 production during vaccine-induced immunity is a requirement for mucosal protection to several agents. In this study, we investigated the potential of a recombinant IL17A to act as an adjuvant for a recombinant BoHV-5 glycoprotein D vaccine in cattle. Three cattle groups were divided as: group 1) rgD5 + alumen + rIL-17A; 2) rgD5 + alumen; and 3) PBS + alumen. The cattle (3 per group) received two doses of their respective vaccines at an interval of 21 days. The group that received rIL17 in its vaccine formulation at the 7th day after the prime immunization had significant higher levels of specific rgD-IgG than the alumen group. Addition of rIL17 also led to a significant fold increase in specific anti-rgD IgG and neutralizing antibodies to the virus, respectively, when compared with the alumen group. Cells stimulated with rIL17A responded with IL17 transcription, as well IL2, IL4, IL10, IL15, Bcl6 and CXCR5. Our findings suggest that the rIL17A has adjuvant potential for use in vaccines against BoHV-5 as well as potentially other pathogens of cattle.
Assuntos
Anticorpos Antivirais/imunologia , Doenças dos Bovinos/prevenção & controle , Encefalite Viral/veterinária , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 5/imunologia , Vacinas contra Herpesvirus/imunologia , Meningoencefalite/veterinária , Adjuvantes Imunológicos , Animais , Anticorpos Neutralizantes/imunologia , Bovinos , Encefalite Viral/prevenção & controle , Infecções por Herpesviridae/prevenção & controle , Herpesvirus Bovino 5/genética , Imunização/veterinária , Interleucina-17/genética , Interleucina-17/imunologia , Meningoencefalite/prevenção & controle , Vacinas Sintéticas , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologiaRESUMO
The aim of this study was to evaluate the humoral immune response in beef heifers supplemented with mineral supplementation with or without the addition of rumen-protected methionine. Forty-eight Brangus nulliparous heifers were distributed into four experimental groups with three replications each: control group without supplementation and without vaccination (CG01), control group without supplementation and with vaccination (CG02), treatment group with mineral supplementation and vaccination (TG01), and treatment group with mineral supplementation added with protected methionine and vaccination (TG02). The animals were maintained under native pasture with access to water ad libitum and the supplementation was available in high-consumption covered troughs. A supplementation period of 60 days prior to vaccinations was adopted until the first dose of a monovalent experimental vaccine inactivated for BoHV-5 was applied as a method of stimulating the immune response to evaluate the supplementation effects. After a 21-day interval, blood samples were collected to evaluate the humoral response and the second vaccine booster dose was applied following the 21-day interval for new blood samples in order to evaluate the immune response against the two-vaccination protocol. From the beginning of the experiment, the animals were weighed on the days −60, −10, 0, 21, and 42 in relation to the vaccine protocol. The experimental groups did not differ for body weight, mean daily weight gain, and body condition score after 102 days of supplementation regardless of the treatment. No animals belonging to CG01 seroconverted throughout the experiment, proving that there was no introduction of the agent (BoHV) in the studied area. When vaccinated animals were compared to the CG01 control group, statistically higher levels of neutralizing antibodies (P ≤ 0.0001) and IgG (P ≤ 0.0001) were verified 21 days after the second vaccine dose...(AU)
O objetivo deste estudo foi avaliar a resposta imune humoral em novilhas de corte suplementadas com suplemento mineral com ou sem adição de metionina protegida da degradação ruminal. Quarenta e oito novilhas nulíparas da raça Brangus, foram distribuídas em quatro grupos experimentais com três repetições cada: Grupo Controle sem suplementação e ausente de vacinação (GC01), Grupo Controle sem suplementação e com vacinação (GC02), Grupo Tratamento com suplementação mineral e com vacinação (GT01) e Grupo Tratamento com suplementação mineral adicionada de metionina protegida e com vacinação (GT02). Os animais foram mantidos em pastagem nativa com acesso à água ad libitum e a suplementação disponibilizada em cochos cobertos de alto consumo. Um período de suplementação prévio às vacinações de 60 dias foi adotado até a aplicação da primeira dose de uma vacina experimental monovalente inativada para BoHV-5, como método de estímulo da resposta imune para avaliar os efeitos da suplementação. Após 21 dias de intervalo, foram realizadas coletas de sangue para avaliação da resposta humoral e aplicada a segunda dose de reforço vacinal, seguindo o intervalo de 21 dias para novas coletas de sangue a fim de avaliar a resposta imune frente ao protocolo de duas vacinações. A partir do início do experimento, os animais foram pesados nos dias -60, -10, 0, 21 e 42 em relação ao protocolo vacinal. Os grupos experimentais, independente do tratamento, não diferiram quanto ao peso corporal, GMD e ECC após os 102 dias de suplementação. Nenhum animal pertencente ao GC01 soroconverteu ao longo do experimento, comprovando que não houve introdução do agente (BoHV) na área estudada. Quando os animais vacinados foram comparados ao grupo controle GC01, níveis estatisticamente superiores de anticorpos neutralizantes (P ≤ 0,0001) e de IgG (P ≤ 0,0001) foram observados 21 dias após a segunda dose vacinal...(AU)
Assuntos
Animais , Feminino , Bovinos , Criação de Animais Domésticos , Herpesvirus Bovino 5/efeitos dos fármacos , Herpesvirus Bovino 5/imunologia , Fatores Imunológicos/sangue , Vacinas/administração & dosagem , Vacinas/sangue , Imunidade Humoral/efeitos dos fármacos , Suplementos Nutricionais , MetioninaRESUMO
Bovine herpesvirus 5 (BoHV-5) is responsible for outbreaks of meningoencephalitis that cause important economic losses in young cattle. BoHV-5 glycoprotein D (gD5) is essential for attachment and penetration into permissive cells and targeting of host immune systems, inducing strong humoral and cellular immune responses. The aim of this study was to evaluate the vaccinal immune response of vaccines formulated with the recombinant BoHV-5 gD (rgD5) in bovines. For the experiment, 72 heifers were randomly allotted into 6 different groups with 12 animals each. Group 1: vaccine formulated using inactivated BoHV-5 (iBoHV-5) adjuvanted with ISA50V2; Group 2: iBoHV-5 associated with 100⯵g of rgD5 adjuvanted with ISA50V2; Group 3: 100⯵g of rgD5 adjuvanted with ISA50V2; Group 4: 100⯵g of rgD5 adjuvanted with Al(OH)3; Group 5: commercial vaccine; and Group 6: control group. Two doses were administered in a 26-day interval and the third after 357â¯days from primo vaccination. Cattle vaccinated with the vaccines formulated with iBoHV-5 plus rgD5 showed a significant (pâ¯<â¯0.01) five-fold increase in total immunoglobulin G (IgG) for BoHV-5, BoHV-1, and rgD5 as compared with the commercial and control groups. Also, a significant (pâ¯<â¯0.05) increase in IgG1 and IgG2a levels was induced in serum for rgD5. In addition, these same vaccines showed significant (pâ¯<â¯0.01) four-fold higher titers of BoHV-1 and -5 neutralizing antibodies. The results demonstrated that the rgD5 conserved important epitopes that were able to stimulate bovine humoral immunity response capable of viral neutralization of BoHV-1 and -5, suggesting it as a promising vaccine antigen to be used in vaccine for BoHV-1 and -5 endemic areas.
Assuntos
Doenças dos Bovinos/prevenção & controle , Infecções por Herpesviridae/veterinária , Proteínas do Envelope Viral/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Antígenos Virais/genética , Antígenos Virais/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Feminino , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/prevenção & controle , Herpesvirus Bovino 1/imunologia , Herpesvirus Bovino 5/imunologia , Esquemas de Imunização , Imunoglobulina G/sangue , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas do Envelope Viral/genética , Vacinas Virais/administração & dosagem , Vacinas Virais/genéticaRESUMO
Bovine herpesvirus (BoHV) types 1 and 5 are neuroinvasive. Cases of BoHV-1-induced encephalitis are not as frequent as those caused by BoHV-5. In this study, the capability of BoHV-5 to induce apoptosis in cell cultures and in the trigeminal ganglion during acute infection of experimentally-infected cattle was analyzed. Apoptotic changes in cell cultures agree with the ability of the viral strains to replicate in each cell line. Marked differences were observed between the in vitro induction of apoptosis by BoHV-1Cooper and BoHV-5 97/613 strains. Apoptotic neurons were clearly evident in the trigeminal ganglion of BoHV-1-infected calves. For BoHV-5 a fewer number of positive neurons was observed. There is an association between the magnitude of bovine herpesviruses replication and the induction of apoptosis in trigeminal ganglion. These findings suggest that the induction of apoptosis and the innate immune response orchestrate the final outcome of alpha herpesviruses infection of the bovine nervous system.
Assuntos
Doenças dos Bovinos/virologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1/patogenicidade , Herpesvirus Bovino 5/patogenicidade , Neurônios/virologia , Gânglio Trigeminal/virologia , Animais , Apoptose , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/patologia , Linhagem Celular , Células Epiteliais/imunologia , Células Epiteliais/patologia , Células Epiteliais/virologia , Células HeLa , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/imunologia , Herpesvirus Bovino 5/imunologia , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Neurônios/imunologia , Neurônios/patologia , Especificidade da Espécie , Gânglio Trigeminal/imunologia , Gânglio Trigeminal/patologia , Replicação ViralRESUMO
There have been significant efforts toward the development of more efficient vaccines for animal health. A strategy that may be used to improve vaccine efficacy is the use of probiotics. Bovine herpesvirus 5 (BoHV-5) is an example of an important animal pathogen for which vaccines have provided only limited protection. In this study, we examined the use of the probiotics Bacillus toyonensis and Saccharomyces boulardii as a potential immune modulator to improve vaccine efficiency. Thirty, 5-month-old lambs were randomly grouped in three lots of 10 each and vaccinated at days 0, 21 and 42 of the experiment. They grazed on the same pasture and were fed ad libitum twice a day with commercial sheep feed supplemented with either B. toyonensis (1×106CFU/g of feed) or S. boulardii (1×107CFU/g of feed), or non-supplemented feed. The probiotic supplementation was suspended day 28; thereafter, the next 35days, they were fed with the same commercial feed as control group. Animals supplemented with probiotics showed a significant (p>0.001) increased seroconversions against BoHV-5, and higher neutralizing antibodies titres (p>0.05) to BoHV-5 than non-supplemented animals. At 63days of experiment, splenocytes from the supplemented sheep had higher mRNA transcription levels of cytokines IL-10 and IL-17A. These results suggest that these probiotics could provide a promising means of improving vaccine efficacy.
Assuntos
Bacillus/imunologia , Herpesvirus Bovino 5/imunologia , Saccharomyces boulardii/imunologia , Vacinas Virais/imunologia , Ração Animal , Animais , Bovinos , Imunomodulação/efeitos dos fármacos , Probióticos/farmacologia , OvinosRESUMO
Probiotics modulate the immune response and can increase the effectiveness of vaccines. Bacillus toyonensis is widely used as a probiotic in animal feed. The aim of this study was to assess the effects of B. toyonensis administration on the immune response to an experimental recombinant vaccine against bovine herpesvirus type 5 (BoHV-5) in mice. Mice were vaccinated with BoHV-5 recombinant glycoprotein D and supplemented with the probiotic B. toyonensis in two regimes: one group received the probiotic only during seven days prior to the initial vaccination while the second group was given the probiotic throughout the experimental period of seven weeks. Animals supplemented with probiotic B. toyonensis in two regimes showed an increase in total immunoglobulin (Ig)G, IgG1 and IgG2a levels in serum, in addition to higher titres of antibodies capable of neutralising the BoHV-5 virus than non-supplemented animals (P<0.05). Splenocytes from the supplemented mice had higher mRNA transcription levels of cytokines interleukin (IL)-4 and IL-12. These results show that the use of this probiotic may significantly contribute to the response elicited by recombinant vaccines, especially those that rely on increasing antibody and cell-mediated immune responses for efficacy. Further, the data support an immunomodulatory effect for probiotic B. toyonensis and imply that enhance effect on the immune response against a BoHV-5 recombinant vaccine in mice.
Assuntos
Bacillus/imunologia , Herpesvirus Bovino 5/imunologia , Vacinas contra Herpesvirus/imunologia , Imunomodulação/efeitos dos fármacos , Probióticos/farmacologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Citocinas/genética , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Vacinas contra Herpesvirus/administração & dosagem , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Fatores Imunológicos/administração & dosagem , Fatores Imunológicos/farmacologia , Camundongos , Probióticos/administração & dosagem , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologiaRESUMO
There have been significant efforts towards the development of more efficient vaccines for animal health. A strategy that may be used to improve vaccine efficacy is the use of probiotics to enhance the immune response of the host, leading to increased immunogenicity of antigen preparations. Bovine herpesvirus 5 (BoHV-5) is an example of an important animal pathogen for which vaccines have provided only limited protection. In this study, we examined the use of the probiotic Saccharomyces boulardii (Sb) as a potential adjuvant to improve vaccine efficiency. We found that the supplemented animals exhibited an enhanced systemic IgG antibody response toward a Th1 response in favor of IgG2a and increased mRNA expression levels of the cytokines IFN-y, IL-12, IL-17 and IL-10 in the spleen. These results suggest that Sb supplementation may provide a promising means for improving the efficiency of vaccines, particularly those that rely on a cell-mediated immune response.(AU)
Esforços significativos têm sido realizados para o desenvolvimento de vacinas mais eficientes em saúde animal. Uma estratégia que pode ser usado para melhorar a eficácia da vacina é o uso de probióticos para melhorar a resposta imune do hospedeiro, conduzindo ao aumento da imunogenicidade de preparações de antígenos. Herpesvírus bovino 5 (BoHV-5) é um exemplo de um importante patógeno animal para os quais vacinas têm fornecido apenas uma protecção limitada. Neste estudo, examinou-se o uso do probiótico Saccharomyces boulardii (Sb) como um adjuvante potencial para melhorar a eficiência da vacina. Verificou-se que os animais suplementados apresentaram uma produção de anticorpos IgG superior e com desvio para Th1 em favor de IgG2, além do aumento dos níveis de expressão de mRNA para as citocinas IFN-γ, IL-12, IL-17 e IL-10. Esses resultados sugerem que a suplementação de Sb pode fornecer um meio promissor para melhorar a eficiência de vacinas, particularmente aquelas que dependem de uma resposta imune mediada por células.(AU)
Assuntos
Encefalite Viral , Infecções por Herpesviridae , Herpesvirus Bovino 5/imunologia , Meningoencefalite , Saccharomyces boulardii/classificaçãoRESUMO
There have been significant efforts towards the development of more efficient vaccines for animal health. A strategy that may be used to improve vaccine efficacy is the use of probiotics to enhance the immune response of the host, leading to increased immunogenicity of antigen preparations. Bovine herpesvirus 5 (BoHV-5) is an example of an important animal pathogen for which vaccines have provided only limited protection. In this study, we examined the use of the probiotic Saccharomyces boulardii (Sb) as a potential adjuvant to improve vaccine efficiency. We found that the supplemented animals exhibited an enhanced systemic IgG antibody response toward a Th1 response in favor of IgG2a and increased mRNA expression levels of the cytokines IFN-y, IL-12, IL-17 and IL-10 in the spleen. These results suggest that Sb supplementation may provide a promising means for improving the efficiency of vaccines, particularly those that rely on a cell-mediated immune response.(AU)
Esforços significativos têm sido realizados para o desenvolvimento de vacinas mais eficientes em saúde animal. Uma estratégia que pode ser usado para melhorar a eficácia da vacina é o uso de probióticos para melhorar a resposta imune do hospedeiro, conduzindo ao aumento da imunogenicidade de preparações de antígenos. Herpesvírus bovino 5 (BoHV-5) é um exemplo de um importante patógeno animal para os quais vacinas têm fornecido apenas uma protecção limitada. Neste estudo, examinou-se o uso do probiótico Saccharomyces boulardii (Sb) como um adjuvante potencial para melhorar a eficiência da vacina. Verificou-se que os animais suplementados apresentaram uma produção de anticorpos IgG superior e com desvio para Th1 em favor de IgG2, além do aumento dos níveis de expressão de mRNA para as citocinas IFN-γ, IL-12, IL-17 e IL-10. Esses resultados sugerem que a suplementação de Sb pode fornecer um meio promissor para melhorar a eficiência de vacinas, particularmente aquelas que dependem de uma resposta imune mediada por células.(AU)
Assuntos
/classificação , Herpesvirus Bovino 5/imunologia , Infecções por Herpesviridae , Encefalite Viral , MeningoencefaliteRESUMO
Bovine herpesvirus type 5 (BoHV-5) is an important pathogen that causes meningoencephalitis in cattle. Few studies have used the mouse as a model for BoHV-5 infection. Despite the fact that BoHV-5 can infect mice with immune deficiencies, little is known about viral replication, immune response, and the course of infection in the central nervous system (CNS) of wild-type mice. Therefore, the aim of this study was to evaluate the response in the CNS of BALB/c mice acutely infected with BoHV-5 at different days post-inoculation (dpi). BoHV-5, when inoculated intracranially, was able to infect and replicate within the CNS of BALB/c mice. Until 15 dpi, the mice were able to survive without showing prominent neurological signs. The infection was accompanied by a Th1 immune response, with a significant expression of the cytokines IFN-γ and TNF-α and chemokine CCL-2. The expression of these cytokines and chemokines was most significant in the early course of infection (3 and 4 dpi), and it was followed by meningoencephalitis with perivascular cuffing and periventriculitis, composed mainly of macrophages and lymphocytes. After the expression of cytokines and chemokine, the mice were able to curb BoHV-5 acute infection in the brain, since there was a decrease in the number of BoHV-5 DNA copies after 3 dpi and viable viral particles were not detected after 6 dpi. Importantly, BoHV-5 was able to infect the trigeminal ganglia during acute infection, since a large number of BoHV-5 DNA copies were detected on 1 and 2 dpi.
Assuntos
Sistema Nervoso Central/imunologia , Suscetibilidade a Doenças/imunologia , Infecções por Herpesviridae/imunologia , Herpesvirus Bovino 5/imunologia , Animais , Sistema Nervoso Central/virologia , Citocinas/sangue , Citocinas/genética , Citocinas/imunologia , Modelos Animais de Doenças , Regulação da Expressão Gênica/imunologia , Herpesvirus Bovino 5/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Gânglio Trigeminal/imunologia , Gânglio Trigeminal/virologia , Replicação ViralRESUMO
Bovine herpesviruses (BoHVs) types 1 (BoHV-1) and 5 (BoHV-5) are alphaherpesviruses of major importance to the bovine production chain. Such viruses are capable of establishing latent infections in neuronal tissues. Infected animals tend to develop a serological response to infection; however, such response-usually investigated by antibody assays in serum-may eventually not be detected in laboratory assays. Nevertheless, serological tests such as virus neutralization (VN) and various enzyme-linked immunosorbent assays (ELISAs) are widely employed to check individual or herd status of BoHV infections. The correlation between detection of antibodies and the presence of viral nucleic acids as indicatives of infection in infected cattle has not been deeply examined. In order to investigate such correlation, 248 bovine serum samples were tested by VN to BoHV-1 and BoHV-5, as well as in a widely employed (though not type-differential) gB ELISA (IDEXX IBR gB X2 Ab Test) in search for antibodies to BoHVs. Immediately after blood withdrawal, cattle were slaughtered and trigeminal ganglia (TG) excised for DNA extraction and viral nucleic acid detection (NAD) by nested PCR. Neutralizing antibodies to BoHV-1 and/or BoHV-5 were detected in 44.8% (111/248) of sera, whereas the gB ELISA detected antibodies in 51.2% (127/248) of the samples. However, genomes of either BoHV-1, BoHV-5, or both, were detected in TGs of 85.9% (213/248) of the animals. These findings reveal that the assays designed to detect antibodies to BoHV-1 and/or BoHV-5 employed here may fail to detect a significant number of latently infected animals (in this study, 35.7%). From such data, it is clear that antibody assays are poorly correlated with detection of viral genomes in BoHV-1 and BoHV-5-infected animals.