RESUMO
Canine distemper virus (CDV) is the causative agent of canine distemper (CD), which is one of the most important infectious diseases affecting wild and domestic carnivores. Vaccination represents an effective approach to prevent CDV infection among domestic carnivores. Canarypox-vectored recombinant CD vaccines (such as Recombitek CDV, PureVax Ferret Distemper, and Merial) with the CDV hemagglutinin (H) and fusion (F) genes can induce a potent immune response in dogs and ferrets. However, the vaccine's effectiveness varies with the species. In the current study, we developed a highly efficient recombinant canarypox virus termed as "ALVAC-CDV-M-F-H/C5-" that contained CDV virus-like particles (VLPs) by using the CRISPR/Cas9 gene editing method, which enabled concurrent expression of the matrix (M), H, and F genes. The recombinant strain provided faster seroconversion than the parent strain among minks as well as provided higher rates of antibody positivity than the parent strain among foxes and minks even before the administration of a second booster vaccination. We demonstrated, for the first time, that the CRISPR/Cas9 system can be applied for the rapid and efficient modification of the ALVAC-CDV-F-H genome and also that a high-dose new recombinant strain that produces CDV VLPs may present good outcomes in the prevention of CD among foxes and minks.
Assuntos
Anticorpos Antivirais/sangue , Sistemas CRISPR-Cas , Vírus da Varíola dos Canários/genética , Vírus da Cinomose Canina/genética , Vírus da Cinomose Canina/imunologia , Cinomose/prevenção & controle , Edição de Genes/métodos , Vacinas Virais/imunologia , Animais , Vírus da Varíola dos Canários/imunologia , Embrião de Galinha/citologia , Galinhas , Chlorocebus aethiops , Cães , Feminino , Fibroblastos/virologia , Raposas/imunologia , Glicoproteínas/genética , Glicoproteínas/imunologia , Hemaglutininas Virais/genética , Hemaglutininas Virais/imunologia , Masculino , Vison/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia , Células Vero , Proteínas Virais de Fusão/genética , Proteínas Virais de Fusão/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/genéticaRESUMO
In the RV144 trial, vaccine-induced V1V2 IgG correlated with decreased HIV-1 risk. We investigated circulating antibody specificities in two phase 1 poxvirus prime-protein boost clinical trials conducted in South Africa: HVTN 097 (subtype B/E) and HVTN 100 (subtype C). With cross-subtype peptide microarrays and multiplex binding assays, we probed the magnitude and breadth of circulating antibody responses to linear variable loop 2 (V2) and conformational V1V2 specificities. Antibodies targeting the linear V2 epitope, a correlate of decreased HIV-1 risk in RV144, were elicited up to 100% and 61% in HVTN 097 and HVTN 100, respectively. Despite higher magnitude of envelope-specific responses in HVTN 100 compared to HVTN 097 (p's < 0.001), the magnitude and positivity for V2 linear epitope and V1V2 proteins were significantly lower in HVTN 100 compared to HVTN 097. Meanwhile, responses to other major linear epitopes including the variable 3 (V3) and constant 5 (C5) epitopes were higher in HVTN 100 compared to HVTN 097. Our data reveal substantial differences in the circulating antibody specificities induced by vaccination in these two canarypox prime-protein boost trials. Our findings suggest that the choice of viral sequences in prime-boost vaccine regimens, and potentially adjuvants and immunogen dose, influence the elicitation of V2-specific antibodies.
Assuntos
Vacinas contra a AIDS/imunologia , Anticorpos Antivirais/imunologia , Formação de Anticorpos/imunologia , HIV-1/imunologia , Especificidade de Anticorpos/imunologia , Vírus da Varíola dos Canários/imunologia , Epitopos/imunologia , Feminino , Proteína gp120 do Envelope de HIV/imunologia , Humanos , Imunização Secundária , MasculinoRESUMO
Viral vectors derived from different virus families, including poxvirus (canarypox virus vector ALVAC) and adenovirus (human Ad5 vector), have been widely used in vaccine development for a range of human diseases including HIV/AIDS. Less is known about the mechanisms underlying the host innate response to these vectors. Increasing evidence from clinical vaccine trials testing different viral vectors has suggested the importance of understanding basic elements of host-viral vector interactions. In this study, we investigated the innate interactions of APCs with two commonly used HIV vaccine vectors, ALVAC and Ad5, and identified AIM2 as an innate sensor for ALVAC, triggering strong inflammasome activation in both human and mouse APCs. Microarray and comprehensive gene-knockout analyses (CRISPR/Cas9) identified that ALVAC stimulated the cGAS/IFI16-STING-type I IFN pathway to prime AIM2, which was functionally required for ALVAC-induced inflammasome activation. We also provided evidence that, in contrast to ALVAC, the Ad5 vector itself was unable to induce inflammasome activation, which was related to its inability to stimulate the STING-type I IFN pathway and to provide inflammasome-priming signals. In preconditioned APCs, the Ad5 vector could stimulate inflammasome activation through an AIM2-independent mechanism. Therefore, our study identifies the AIM2 inflammasome and cGAS/IFI16-STING-type I IFN pathway as a novel mechanism for host innate immunity to the ALVAC vaccine vector.
Assuntos
Adenoviridae/imunologia , Células Apresentadoras de Antígenos/imunologia , Vírus da Varíola dos Canários/imunologia , Proteínas de Ligação a DNA/imunologia , Vetores Genéticos/imunologia , Imunidade Inata , Interferon Tipo I/imunologia , Proteínas de Membrana/imunologia , Proteínas Nucleares/imunologia , Nucleotidiltransferases/imunologia , Fosfoproteínas/imunologia , Transdução de Sinais/imunologia , Animais , Sistemas CRISPR-Cas , Proteínas de Ligação a DNA/genética , Feminino , Técnicas de Silenciamento de Genes , Humanos , Interferon Tipo I/genética , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Knockout , Proteínas Nucleares/genética , Nucleotidiltransferases/genética , Fosfoproteínas/genética , Transdução de Sinais/genéticaRESUMO
Canarypox viruses (CNPV) are excellent candidates to develop recombinant vector vaccines due to both their capability to induce protective immune responses and their incompetence to replicate in mammalian cells (safety profile). In addition, CNPV and the derived recombinants can be manipulated under biosafety level 1 conditions. There is no commercially available system to obtain recombinant CNPV; however, the methodology and tools required to develop recombinant vaccinia virus (VV), prototype of the Poxviridae family, can be easily adapted. This chapter provides protocols for the generation, plaque isolation, molecular characterization, amplification and purification of recombinant CNPV.
Assuntos
Vírus da Varíola dos Canários/crescimento & desenvolvimento , Fibroblastos/virologia , Vacinas Sintéticas/imunologia , Animais , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/imunologia , Linhagem Celular , Galinhas , Fibroblastos/imunologia , Vacinas Virais/imunologiaAssuntos
Infecções por Birnaviridae/veterinária , Vírus da Varíola dos Canários/imunologia , Vírus da Doença Infecciosa da Bursa/genética , Doenças das Aves Domésticas/imunologia , Proteínas Estruturais Virais/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Antígenos Virais/genética , Antígenos Virais/imunologia , Infecções por Birnaviridae/imunologia , Infecções por Birnaviridae/prevenção & controle , Bolsa de Fabricius/diagnóstico por imagem , Bolsa de Fabricius/patologia , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/metabolismo , Galinhas , Vetores Genéticos , Doenças das Aves Domésticas/prevenção & controle , Doenças das Aves Domésticas/virologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas Estruturais Virais/biossíntese , Proteínas Estruturais Virais/genética , Vacinas Virais/administração & dosagem , Vacinas Virais/genéticaRESUMO
Despite the growing use of poxvirus vectors as vaccine candidates for multiple pathogens and cancers, their innate stimulatory properties remain poorly characterized. Here we show that the canarypox virus-based vector ALVAC induced distinct systemic proinflammatory and antiviral cytokine and chemokine levels following the vaccination of rhesus monkeys compared to the vaccinia virus-based vectors MVA and NYVAC. These data suggest that there are substantial biological differences among leading poxvirus vaccine vectors that may influence resultant adaptive immune responses following vaccination.
Assuntos
Vírus da Varíola dos Canários/imunologia , Citocinas/imunologia , Vetores Genéticos/imunologia , Vaccinia virus/imunologia , Vacinas Virais/imunologia , Animais , Vírus da Varíola dos Canários/genética , Vetores Genéticos/genética , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/imunologia , HIV-1/fisiologia , Humanos , Macaca mulatta , Proteínas dos Retroviridae/administração & dosagem , Proteínas dos Retroviridae/genética , Proteínas dos Retroviridae/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/imunologia , Vírus da Imunodeficiência Símia/fisiologia , Vaccinia virus/genética , Vacinas Virais/administração & dosagem , Vacinas Virais/genéticaRESUMO
Morbilliviruses pose a significant threat to marine mammal populations around the world and have been associated with multiple epizootics in pinnipeds and cetaceans. As part of a preventive veterinary medical program, five harbor seals (Phoca vitulina) in a managed collection were vaccinated with a recombinant canarypox-vectored canine distemper virus vaccine. The vaccine was evaluated for safety (by monitoring seals for local and systemic adverse effects and by testing for shedding of the canarypox vector) and efficacy (by testing for serum neutralizing antibodies). None of the seals showed signs of local or systemic adverse reactions to the vaccination. Three seals vaccinated once did not seroconvert, but the recombinant vaccine induced a persistent serum virus neutralizing titer (12 mo) in the two seals that were vaccinated twice, 1 mo apart.
Assuntos
Vírus da Cinomose Focina/imunologia , Infecções por Morbillivirus/veterinária , Phoca , Vacinas Virais/administração & dosagem , Animais , Anticorpos Antivirais/sangue , Vírus da Varíola dos Canários/imunologia , Vírus da Cinomose Canina/imunologia , Morbillivirus , Infecções por Morbillivirus/prevenção & controle , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/efeitos adversos , Vacinas Virais/efeitos adversosRESUMO
Development and preliminary assessment of a recombinant canarypox virus as an antirabic vaccine candidate. In Argentina, rabies is limited to some northern provinces. Availability of new vaccines abolishing the handling of the rabies virus and allowing disease control has regional and national strategic importance. Vaccines based on recombinant poxviruses have been successfully used as antirabic vaccines worldwide. Although these systems are not commercially available, the platform to obtain recombinant canarypox viruses (CNPV) has been previously set up in our laboratory. The aim of this work was the development and evaluation of an antirabic vaccine candidate based on recombinant CNPV expressing the rabies virus (RV) glycoprotein G (RG). A recombinant virus (CNPV-RG) expressing the RG coding sequence was designed. Inoculation of mice with this virus induced high RV seroneutralizing antibodies (3.58 and 9.76 IU/ml after 1 or 2 immunizations, respectively) and protected 78% of intracerebrally RV-challenged animals. In addition, it was determined that CNPV-RG has a relative potency of 3.5 IU/ml. The obtained results constituted the first stage of CNPV-RG evaluation as antirabic vaccine candidate. Further assays will be necessary to confirm its utility in species of veterinary interest.
Assuntos
Antígenos Virais/imunologia , Vírus da Varíola dos Canários/imunologia , Glicoproteínas/imunologia , Vacina Antirrábica , Proteínas do Envelope Viral/imunologia , Animais , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/imunologia , Antígenos Virais/genética , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/crescimento & desenvolvimento , Vírus da Varíola dos Canários/isolamento & purificação , Linhagem Celular/virologia , Embrião de Galinha , Chlorocebus aethiops , Cricetinae , Fibroblastos/virologia , Glicoproteínas/genética , Rim , Mesocricetus , Camundongos , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Raiva/prevenção & controle , Vacina Antirrábica/imunologia , Organismos Livres de Patógenos Específicos , Vacinas Sintéticas/imunologia , Células Vero/virologia , Proteínas do Envelope Viral/genética , Cultura de VírusRESUMO
En la Argentina, la rabia está circunscripta a algunas provincias del norte. La disponibilidad de nuevas vacunas que eliminen la manipulación del virus rábico y que permitan el control de la enfermedad es de importancia estratégica nacional y regional. Las vacunas basadas en poxvirus recombinantes se han utilizado con éxito como vacunas antirrábicas a nivel mundial. SI bien estos sistemas no están disponibles comercialmente, la plataforma de obtención de virus canarypox (CNPV) recombinantes ya ha sido implementada en nuestro laboratorio. El objetivo de este trabajo fue obtener y evaluar un candidato a vacuna antirrábica basado en CNPV recombinantes que expresan la glicoproteína G (RG) del virus rábico (RV). Se construyó un virus recombinante que expresa la secuencia codificante de RG (CNPV-RG). La inoculación de ratones con este virus indujo altos títulos de anticuerpos seroneutralizantes de RV (3,58 y 9,76 Ul/ml después de una o dos inmunizaciones, respectivamente) y protegió al 78 % de los animales desafiados intracerebralmente con RV. Además, se determinó que el CNPV-RG posee una potencia relativa de 3,5 Ul/ml. Los resultados obtenidos constituyen la primera etapa en la evaluación del CNPV-RG como candidato a vacuna antirrábica. Se requerirán nuevos ensayos para confirmar su utilidad en especies de interés veterinario.
In Argentina, rabies is limited to some northern provinces. Availability of new vaccines abolishing the handling of the rabies virus and allowing disease control has regional and national strategic importance. Vaccines based on recombinant poxviruses have been successfully used as antirabic vaccines worldwide. Although these systems are not commercially available, the platform to obtain recombinant canarypox viruses (CNPV) has been previously set up in our laboratory. The aim of this work was the development and evaluation of an antirabic vaccine candidate based on recombinant CNPV expressing the rabies virus (RV) glycoprotein G (RG). A recombinant virus (CNPV-RG) expressing the RG coding sequence was designed. Inoculation of mice with this virus induced high RV seroneutralizing antibodies (3.58 and 9.76 lU/ml after 1 or 2 immunizations, respectively) and protected 78% of intracerebrally RV-challenged animals. In addition, it was determined that CNPV-RG has a relative potency of 3.5 lU/ml. The obtained results constituted the first stage of CNPV-RG evaluation as antirabic vaccine candidate. Further assays will be necessary to confirm its utility in species of veterinary Interest.
Assuntos
Animais , Embrião de Galinha , Cricetinae , Camundongos , Antígenos Virais/imunologia , Vírus da Varíola dos Canários/imunologia , Glicoproteínas/imunologia , Vacina Antirrábica , Proteínas do Envelope Viral/imunologia , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/imunologia , Antígenos Virais/genética , Chlorocebus aethiops , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/crescimento & desenvolvimento , Vírus da Varíola dos Canários/isolamento & purificação , Linhagem Celular/virologia , Fibroblastos/virologia , Glicoproteínas/genética , Rim , Mesocricetus , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Vacina Antirrábica/imunologia , Raiva/prevenção & controle , Organismos Livres de Patógenos Específicos , Cultura de Vírus , Vacinas Sintéticas/imunologia , Células Vero/virologia , Proteínas do Envelope Viral/genéticaRESUMO
En la Argentina, la rabia está circunscripta a algunas provincias del norte. La disponibilidad de nuevas vacunas que eliminen la manipulación del virus rábico y que permitan el control de la enfermedad es de importancia estratégica nacional y regional. Las vacunas basadas en poxvirus recombinantes se han utilizado con éxito como vacunas antirrábicas a nivel mundial. SI bien estos sistemas no están disponibles comercialmente, la plataforma de obtención de virus canarypox (CNPV) recombinantes ya ha sido implementada en nuestro laboratorio. El objetivo de este trabajo fue obtener y evaluar un candidato a vacuna antirrábica basado en CNPV recombinantes que expresan la glicoproteína G (RG) del virus rábico (RV). Se construyó un virus recombinante que expresa la secuencia codificante de RG (CNPV-RG). La inoculación de ratones con este virus indujo altos títulos de anticuerpos seroneutralizantes de RV (3,58 y 9,76 Ul/ml después de una o dos inmunizaciones, respectivamente) y protegió al 78 % de los animales desafiados intracerebralmente con RV. Además, se determinó que el CNPV-RG posee una potencia relativa de 3,5 Ul/ml. Los resultados obtenidos constituyen la primera etapa en la evaluación del CNPV-RG como candidato a vacuna antirrábica. Se requerirán nuevos ensayos para confirmar su utilidad en especies de interés veterinario.(AU)
In Argentina, rabies is limited to some northern provinces. Availability of new vaccines abolishing the handling of the rabies virus and allowing disease control has regional and national strategic importance. Vaccines based on recombinant poxviruses have been successfully used as antirabic vaccines worldwide. Although these systems are not commercially available, the platform to obtain recombinant canarypox viruses (CNPV) has been previously set up in our laboratory. The aim of this work was the development and evaluation of an antirabic vaccine candidate based on recombinant CNPV expressing the rabies virus (RV) glycoprotein G (RG). A recombinant virus (CNPV-RG) expressing the RG coding sequence was designed. Inoculation of mice with this virus induced high RV seroneutralizing antibodies (3.58 and 9.76 lU/ml after 1 or 2 immunizations, respectively) and protected 78% of intracerebrally RV-challenged animals. In addition, it was determined that CNPV-RG has a relative potency of 3.5 lU/ml. The obtained results constituted the first stage of CNPV-RG evaluation as antirabic vaccine candidate. Further assays will be necessary to confirm its utility in species of veterinary Interest.(AU)
Assuntos
Animais , Embrião de Galinha , Cricetinae , Camundongos , Antígenos Virais/imunologia , Vírus da Varíola dos Canários/imunologia , Glicoproteínas/imunologia , Vacina Antirrábica , Proteínas do Envelope Viral/imunologia , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/imunologia , Antígenos Virais/genética , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/crescimento & desenvolvimento , Vírus da Varíola dos Canários/isolamento & purificação , Linhagem Celular/virologia , Chlorocebus aethiops , Fibroblastos/virologia , Glicoproteínas/genética , Rim , Mesocricetus , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Raiva/prevenção & controle , Vacina Antirrábica/imunologia , Organismos Livres de Patógenos Específicos , Vacinas Sintéticas/imunologia , Células Vero/virologia , Proteínas do Envelope Viral/genética , Cultura de VírusRESUMO
Vaccine strategies aimed at generating CD8(+) T cell memory responses are likely to show augmented efficacy against chronic challenges like tumor. The abundance in variety of memory CD8(+) T cells behooves development of vaccine strategies that generate distinct memory responses and evaluate them for tumor efficacy. In this study, we demonstrate the ability of a variety of rapamycin treatment regimens to regulate virus vaccination-induced CD8(+) T cell memory responses and tumor efficacy. Strikingly, a short course of high-dose, but not low-dose, rapamycin treatment transiently blocks viral vaccination-induced mammalian target of rapamycin activity in CD8(+) T cells favoring persistence and Ag-recall responses over type 1 effector maturation; however, prolonged high-dose rapamycin administration abrogated memory responses. Furthermore, a short course of high-dose rapamycin treatment generated CD8(+) T cell memory responses that were independent of IL-15 and IL-7 and were programmed early for sustenance and greater tumor efficacy. These results demonstrate the impact a regimen of rapamycin treatment has on vaccine-induced CD8(+) T cell responses and indicates that judicious application of rapamycin can augment vaccine efficacy for chronic challenges.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/uso terapêutico , Memória Imunológica/efeitos dos fármacos , Proteínas/imunologia , Sirolimo/uso terapêutico , Timoma/imunologia , Neoplasias do Timo/imunologia , Vacinação , Adjuvantes Imunológicos/farmacologia , Transferência Adotiva , Animais , Antígenos de Neoplasias/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/transplante , Vírus da Varíola dos Canários/imunologia , Vacinas Anticâncer/administração & dosagem , Seleção Clonal Mediada por Antígeno , Interleucina-15/deficiência , Interleucina-15/imunologia , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Complexos Multiproteicos , Transplante de Neoplasias , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Sirolimo/farmacologia , Especificidade do Receptor de Antígeno de Linfócitos T , Serina-Treonina Quinases TOR , Timoma/terapia , Neoplasias do Timo/terapiaRESUMO
Vaccines may be effective tools for protecting small populations of highly susceptible endangered, captive-reared, or translocated Hawaiian honeycreepers from introduced Avipoxvirus, but their efficacy has not been evaluated. An attenuated Canarypox vaccine that is genetically similar to one of two passerine Avipoxvirus isolates from Hawai'i and distinct from Fowlpox was tested to evaluate whether Hawai'i 'Amakihi (Hemignathus virens) can be protected from wild isolates of Avipoxvirus from the Hawaiian Islands. Thirty-one (31) Hawai'i 'Amakihi were collected from high-elevation habitats on Mauna Kea Volcano, where pox transmission is rare, and randomly divided into two groups. One group was vaccinated with Poximune C, whereas the other group received a sham vaccination with sterile water. Four of 15 (27%) vaccinated birds developed life-threatening disseminated lesions or lesions of unusually long duration, whereas one bird never developed a vaccine-associated lesion or "take." After vaccine lesions healed, vaccinated birds were randomly divided into three groups of five and challenged with either a wild isolate of Fowlpox (FP) from Hawai'i, a Hawai'i 'Amakihi isolate of a Canarypox-like virus (PV1), or a Hawai'i 'Amakihi isolate of a related, but distinct, passerine Avipoxvirus (PV2). Similarly, three random groups of five unvaccinated 'Amakihi were challenged with the same virus isolates. Vaccinated and unvaccinated 'Amakihi challenged with FP had transient infections with no clinical signs of infection. Mortality in vaccinated 'Amakihi challenged with PV1 and PV2 ranged from 0% (0/5) for PV1 to 60% (3/5) for PV2. Mortality in unvaccinated 'Amakihi ranged from 40% (2/5) for PV1 to 100% (5/5) for PV2. Although the vaccine provided some protection against PV1, both potential for vaccine reversion and low efficacy against PV2 preclude its use in captive or wild honeycreepers.
Assuntos
Vírus da Varíola dos Canários/imunologia , Passeriformes , Infecções por Poxviridae/veterinária , Vacinas Virais/imunologia , Animais , Vírus da Varíola dos Canários/patogenicidade , Espécies em Perigo de Extinção , Regulação Viral da Expressão Gênica , Havaí/epidemiologia , Filogenia , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/prevenção & controle , Infecções por Poxviridae/virologia , Vacinas Atenuadas , Proteínas do Core Viral/genética , Proteínas do Core Viral/metabolismo , VirulênciaRESUMO
The results of an accelerated immunisation schedule for horses used as part of the emergency response plan to contain and eradicate equine influenza in Australia in 2007 is described. The horses studied were vaccinated with a recombinant canarypox-vectored vaccine (ProteqFlu®, Merial) with a shorter interdose interval. Vaccinated horses included foals aged less than 4 months.
Assuntos
Surtos de Doenças/veterinária , Doenças dos Cavalos/prevenção & controle , Doenças dos Cavalos/virologia , Vírus da Influenza A Subtipo H3N8/imunologia , Vacinas contra Influenza/administração & dosagem , Infecções por Orthomyxoviridae/veterinária , Animais , Animais Recém-Nascidos , Anticorpos Antivirais/sangue , Austrália/epidemiologia , Vírus da Varíola dos Canários/imunologia , Surtos de Doenças/prevenção & controle , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/imunologia , Cavalos , Vírus da Influenza A Subtipo H3N8/genética , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , RNA Viral/química , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Estudos Soroepidemiológicos , Vacinação/métodos , Vacinação/normas , Vacinação/veterinária , Vacinas Sintéticas/imunologiaRESUMO
Highly attenuated poxviruses are promising vectors for protective and therapeutic vaccines. These vectors do not replicate in human cells and can therefore be safely given even to immunocompromised recipients. They can accommodate very large inserts and provide strong stimulation of the immune system against the vectored antigen. Disadvantages include that very high numbers of infectious units are required per dose for full efficacy. Because they are difficult to produce, improved cellular substrates and processes are urgently needed to facilitate programs intended to reach a large number of vaccinees. We have developed a fully scalable and very efficient chemically-defined production process for modified vaccinia Ankara (MVA), canarypox (CNPV, strain ALVAC) and fowlpox viruses (FPV) based on a continuous cell line.
Assuntos
Vetores Genéticos/genética , Poxviridae/genética , Animais , Reatores Biológicos , Células CHO , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/imunologia , Linhagem Celular , Proliferação de Células , Cricetinae , Cricetulus , Vírus da Varíola das Aves Domésticas/genética , Vírus da Varíola das Aves Domésticas/imunologia , Vetores Genéticos/imunologia , Humanos , Poxviridae/imunologia , Vacinas Atenuadas/imunologia , Vaccinia virus/genética , Vaccinia virus/imunologia , Vacinas Virais/imunologia , Replicação Viral/genéticaRESUMO
BACKGROUND: HIV-specific T-cell-based vaccines have been extensively studied in both prevention and therapeutic settings, with most studies failing to show benefit, and some suggesting harm. We previously performed a multicenter, double-blind, placebo-controlled phase II clinical trial in which 65 antiretroviral-treated patients were randomized to receive an HIV-1 recombinant canarypox vaccine (vCP1452) or placebo, followed by analytical treatment interruption. Patients exposed to vaccine had higher levels of viral replication and more rapid time to treatment resumption. OBJECTIVE: In the present study we report the results from extensive immunological investigations to test whether the preferential expansion of HIV-specific CD4(+), rather than CD8(+) T cells, could account for these unexpected results. METHODS: Polychromatic flow cytometry was used to characterize the functional and phenotypic profile of antigen-specific CD8(+) and CD4(+) T cells induced by the immunization. RESULTS: We found a significant increase in HIV-specific CD4(+) T cells producing IFN-γ and IL-2 in the 4 injections arm compared to the placebo arm following vaccination. In contrast, no difference was observed following vaccination in the phenotype and functional capacity within the CD8(+) T-cell compartment. Neither HLA biases, nor immune hyper-activation, or Env-specific facilitating antibodies were associated with the enhanced virus rebound observed in vaccinees. CONCLUSION: Our data suggest that a vaccine-induced transient activation of HIV-specific CD4(+) but not CD8(+) T cells may have a detrimental effect on HIV outcomes. These findings may provide a mechanistic basis for higher rates of HIV acquisition or replication that have been associated with some T-cell vaccines.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Infecções por HIV/imunologia , HIV-1 , RNA Viral/imunologia , Vacinas Virais/uso terapêutico , Anticorpos Neutralizantes/imunologia , Vírus da Varíola dos Canários/imunologia , Método Duplo-Cego , Infecções por HIV/terapia , Infecções por HIV/virologia , Humanos , RNA Viral/efeitos dos fármacos , Carga Viral , Vacinas Virais/imunologiaRESUMO
Equine influenza virus is a leading cause of respiratory disease in the horse population; however, the susceptibility of old horses to EIV infection remains unknown. While advanced age in horses (>20 years) is associated with age-related changes in immune function, there are no specific recommendations regarding the vaccination of older horses even though a well-characterized effect of aging is a reduced antibody response to standard vaccination. Therefore, we evaluated the immunological and physiological response of aged horses to a live non-replicating canarypox-vectored EIV vaccine and subsequent challenge infection. Vaccination of the aged horses induced EIV-specific IgGb and HI antibodies. No specific increase in cell-mediated immune (CMI) response was induced by the vaccine as determined by EIV-specific lymphoproliferation and the detection of EIV-specific IFNγ(+) CD5(+)T cells, IFNγ, IL-2, IL-4 and IL-13 mRNA expression. Non-vaccinated aged horses exhibited clinical signs of the disease (coughing, nasal discharge, dyspnea, depression, anorexia) as well as increased rectal temperature and viral shedding following challenge. Concomitant with the febrile episodes, we also observed increased production of pro-inflammatory cytokine mRNA production in vivo using RT-PCR. Naïve horses were included in this study for vaccine and challenge controls only. As expected, the canarypox-vectored EIV vaccine stimulated significant CMI and humoral immune responses and provided significant protection against clinical signs of disease and reduced virus shedding in naive horses. Here, we show that aged horses remain susceptible to infection with equine influenza virus despite the presence of circulating antibodies and CMI responses to EIV and vaccination with a canarypox-vectored EIV vaccine provides protection from clinical disease.
Assuntos
Envelhecimento/imunologia , Vírus da Varíola dos Canários/imunologia , Doenças dos Cavalos/prevenção & controle , Vírus da Influenza A Subtipo H3N8/imunologia , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/imunologia , Animais , Cavalos , Imunidade Celular , Imunidade Humoral , Vacinas Sintéticas/imunologiaRESUMO
Canarypox and fowlpox viruses represent alternative vaccine vectors due to their natural host-range restriction to avian species. Although they cannot replicate in mammals, they correctly express transgenes in human cells and elicit a complete immune response in vaccinated subjects. Several studies have evaluated their genomic differences and protective efficacy in preclinical trials, but detailed information is not available for their transgene expression, cytokine modulation and abortive replication in mammals. This study demonstrates that the heterologous HIV gag/pol and env genes are more efficiently expressed by fowlpox in non-immune and immune cells. The production of retrovirus-like particles, the longer transgene expression, and a balanced cytokine induction may confer to fowlpox-based recombinants the ability to elicit a better immune response.
Assuntos
Vacinas contra a AIDS , Vírus da Varíola dos Canários , Vírus da Varíola das Aves Domésticas , Vetores Genéticos , HIV-1/genética , Vacinas Sintéticas , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Animais , Antígeno B7-1/análise , Vírus da Varíola dos Canários/genética , Vírus da Varíola dos Canários/imunologia , Vírus da Varíola dos Canários/fisiologia , Vírus da Varíola dos Canários/ultraestrutura , Linhagem Celular , Citocinas/imunologia , Células Dendríticas/imunologia , Imunofluorescência , Vírus da Varíola das Aves Domésticas/genética , Vírus da Varíola das Aves Domésticas/imunologia , Vírus da Varíola das Aves Domésticas/fisiologia , Vírus da Varíola das Aves Domésticas/ultraestrutura , Expressão Gênica , Genes env , Genes gag , Genes pol , HIV-1/imunologia , Humanos , Imunização , Macrófagos/imunologia , Microscopia Eletrônica de Transmissão , Transgenes , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Replicação Viral/genéticaRESUMO
Several felid species have been shown to be susceptible to infection with highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype. Infection of felids by H5N1 HPAI virus is often fatal, and cat-to-cat transmission has been documented. Domestic cats may then be involved in the transmission of infection to other animals but also to humans. A particular concern is the hypothetical role of the cat in the adaptation of the virus to mammalian species, thus increasing the pandemic risk. Therefore, the development of a HPAI vaccine for domestic cats should be considered a veterinary and also a public health priority. Here we show that vaccination of cats with a recombinant canarypox (ALVAC)(1)) virus, expressing the hemagglutinin (HA) of influenza virus A/chicken/Indonesia/03 (H5N1) confers protection against challenge infection with two antigenically distinct H5N1 virus isolates from humans. Despite low hemagglutination inhibiting (HI) antibody titers at the time of challenge, all vaccinated cats were protected against mortality and had reduced histopathological changes in the lungs. Importantly, viral shedding was reduced in vaccinated cats as compared to controls, suggesting that vaccination of cats could reduce the risk of viral transmission. In conclusion this study showed that the recombinant canarypox virus protected cats against homologous and heterologous H5N1 HPAI virus challenges.
Assuntos
Proteção Cruzada , Virus da Influenza A Subtipo H5N1/imunologia , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/veterinária , Animais , Anticorpos Antivirais/sangue , Formação de Anticorpos , Vírus da Varíola dos Canários/imunologia , Gatos , Testes de Inibição da Hemaglutinação , Virus da Influenza A Subtipo H5N1/fisiologia , Pulmão/patologia , Pulmão/virologia , Infecções por Orthomyxoviridae/imunologia , Eliminação de Partículas ViraisAssuntos
Vacinas contra a AIDS/imunologia , Infecções por HIV/prevenção & controle , HIV/imunologia , Anticorpos Neutralizantes/análise , Anticorpos Neutralizantes/imunologia , Vírus da Varíola dos Canários/imunologia , Vírus da Varíola dos Canários/metabolismo , Ensaios Clínicos como Assunto , HIV/patogenicidade , Anticorpos Anti-HIV/análise , Anticorpos Anti-HIV/imunologia , Proteína gp120 do Envelope de HIV/imunologia , Infecções por HIV/imunologia , Infecções por HIV/virologia , Humanos , Imunidade Celular , Imunização Secundária , Tailândia/epidemiologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologiaRESUMO
Due to their natural host-range restriction to avian species, canarypox virus (CP) and fowlpox virus (FP) represent efficient and safe vaccine vectors, as they correctly express transgenes in human cells, elicit complete immune responses, and show protective efficacy in preclinical animal models. At present, no information is available on the differences in the abortive replication of these two avipox viruses in mammalian cells. In the present study, the replicative cycles of CP and FP, wild-type and recombinants, are compared in permissive and non-permissive cells, using transmission electron microscopy. We demonstrate that in non-permissive cells, the replicative cycle is more advanced in FP than in CP, that human cells, whether immune or not, are less permissive to avipox replication than monkey cells, and that the presence of virus-like particles only occurs after FP infection. Overall, these data suggest that the use of FP recombinants is more appropriate than the use of CP for eliciting an immune response.