RESUMEN
Seven colostrum-deprived, 3-4-wk-old Rambouillet-Hampshire lambs were inoculated via the mucous membranes with deer adenovirus (DAdV) and monitored for clinical signs for 21 d post-inoculation at which time animals were euthanized and postmortem examinations were performed. Pre-inoculation and post-inoculation serum samples were tested for antibodies to DAdV, ovine adenovirus 7, bovine adenovirus 7, and goat adenovirus 1. Evidence for DAdV infection was determined by virus isolation, PCR tests, and histopathology with immunohistochemistry tests for DAdV. No clinical signs or lesions consistent with adenoviral hemorrhagic disease (AHD) in deer were seen in the lambs, and the lambs did not seroconvert to DAdV. DAdV was not detected by PCR, virus isolation, or immunohistochemistry in any of the samples tested from the lambs. A positive control deer similarly inoculated with DAdV developed fatal AHD 1 wk post-inoculation. Our colostrum-deprived lambs did not become infected when inoculated with DAdV.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Atadenovirus/aislamiento & purificación , Calostro/inmunología , Enfermedades de las Ovejas/virología , Infecciones por Adenoviridae/inmunología , Animales , Animales Domésticos , Animales Recién Nacidos , Animales Lactantes , Atadenovirus/inmunología , Femenino , Inmunohistoquímica/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Embarazo , Ovinos , Enfermedades de las Ovejas/inmunologíaRESUMEN
Identification of adenoviral isolates of nonhuman origin has fostered development of vectors with potential to overcome preexisting immunity in the human population that may affect clinical applications. Ovine adenoviral isolate, OAdV287 (OAdV7), the prototype of the genus Atadenovirus, has been previously characterized as a gene delivery vector although the receptor(s) used for infection remain to be identified. Here, we report the first use of recombinant OAdV7 as a vaccine for inducing an antitumor immune response in a mouse model. Treatment of murine BMDC with OAdV7 vectors expressing ovalbumin (OVA) resulted in upregulation of costimulatory markers and production of IL-12. Splenocytes isolated from immunized mice responded to antigen restimulation in vitro by proliferation and production of IFNγ. In vivo cytotoxicity assays revealed efficient killing of antigenic peptide-pulsed target cells 1 week after immunization, with an average killing efficiency of 75%. In mice inoculated with B16-OVA tumor cells immunization with OAdV7-OVA retarded and essentially prevented tumor growth in prophylactic and therapeutic tumor trials, respectively. Generation of a robust memory response was confirmed on tumor rechallenge in the prophylactic model. Therefore, OAdV7 is a novel vector with potential for further development of tumor vaccines.
Asunto(s)
Atadenovirus/inmunología , Células Dendríticas/metabolismo , Vectores Genéticos , Melanoma Experimental/inmunología , Melanoma Experimental/terapia , Animales , Antígenos de Neoplasias/inmunología , Diferenciación Celular/inmunología , Proliferación Celular , Células Dendríticas/inmunología , Células Dendríticas/patología , Células Dendríticas/virología , Inmunización , Memoria Inmunológica , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Ovalbúmina/genética , Ovalbúmina/inmunología , Ovinos , Carga TumoralRESUMEN
The desire to induce HIV-1-specific responses soon after birth to prevent breast milk transmission of HIV-1 led us to propose a vaccine regimen which primes HIV-1-specific T cells using a recombinant Mycobacterium bovis bacillus Calmette-Guérin (rBCG) vaccine. Because attenuated live bacterial vaccines are typically not sufficiently immunogenic as stand-alone vaccines, rBCG-primed T cells will likely require boost immunization(s). Here, we compared modified Danish (AERAS-401) and Pasteur lysine auxotroph (222) strains of BCG expressing the immunogen HIVA for their potency to prime HIV-1-specific responses in adult BALB/c mice and examined four heterologous boosting HIVA vaccines for their immunogenic synergy. We found that both BCG.HIVA(401) and BCG.HIVA(222) primed HIV-1-specific CD8(+) T-cell-mediated responses. The strongest boosts were delivered by human adenovirus-vectored HAdV5.HIVA and sheep atadenovirus-vectored OAdV7.HIVA vaccines, followed by poxvirus MVA.HIVA; the weakest was plasmid pTH.HIVA DNA. The prime-boost regimens induced T cells capable of efficient in vivo killing of sensitized target cells. We also observed that the BCG.HIVA(401) and BCG.HIVA(222) vaccines have broadly similar immunologic properties, but display a number of differences mainly detected through distinct profiles of soluble intercellular signaling molecules produced by immune splenocytes in response to both HIV-1- and BCG-specific stimuli. These results encourage further development of the rBCG prime-boost regimen.
Asunto(s)
Vacunas contra el SIDA/inmunología , Vacuna BCG/inmunología , Linfocitos T CD8-positivos/inmunología , Epítopos de Linfocito T/inmunología , VIH-1/inmunología , Vacunas Virales/inmunología , Adenoviridae/inmunología , Animales , Atadenovirus/inmunología , Femenino , Vectores Genéticos/inmunología , Infecciones por VIH/inmunología , Inmunización/métodos , Inmunización Secundaria/métodos , Ratones , Ratones Endogámicos BALB C , Ovinos , Transducción de Señal/inmunología , Vacunación/métodos , Vacunas Sintéticas/inmunologíaAsunto(s)
Anticuerpos Antivirales/sangre , Ciervos/virología , Vigilancia de Guardia/veterinaria , Infecciones por Adenoviridae/epidemiología , Infecciones por Adenoviridae/veterinaria , Animales , Animales Domésticos/virología , Animales Salvajes/virología , Atadenovirus/inmunología , Diarrea Mucosa Bovina Viral/epidemiología , Bovinos , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/veterinaria , Coronavirus Bovino/inmunología , Virus de la Diarrea Viral Bovina/inmunología , Japón/epidemiología , Estudios SeroepidemiológicosRESUMEN
Ovine adenovirus type 7 (OAdV) is the prototype member of the genus Atadenovirus. No immunity to the virus has so far been detected in human sera. We describe the construction and evaluation of a candidate HIV-1 vaccine based on OAdV and its utilisation alone and in combination with plasmid-, human adenovirus type 5 (HAdV5; a Mastadenovirus)-, and modified vaccinia Ankara (MVA)-vectored vaccines. All vectors expressed HIVA, an immunogen consisting of HIV-1 clade A consensus Gag-derived protein coupled to a T cell polyepitope. OAdV.HIVA was genetically stable, grew well and expressed high levels of protein from the Rous sarcoma virus promoter. OAdV.HIVA was highly immunogenic in mice and efficiently primed and boosted HIV-1-specific T cell responses together with heterologous HIVA-expressing vectors. There were significant differences between OAdV and HAdV5 vectors in priming of naïve CD8(+) T cell responses to HIVA and in the persistence of MHC class I-restricted epitope presentation in the local draining lymph nodes. OAdV.HIVA primed T cells more rapidly but was less persistent than AdV5.HIVA and thus induced a qualitatively distinct T cell response. Nevertheless, both vectors primed a response in mice that reduced viral titres in a surrogate challenge model by three to four orders of magnitude. Thus, OAdV is a novel, underexplored vaccine vector with potential for further development for HIV-1 and other vaccines. The data are discussed in the context of the latest HIV-1 vaccine developments.
Asunto(s)
Vacunas contra el SIDA/genética , Vacunas contra el SIDA/inmunología , Atadenovirus/genética , Atadenovirus/inmunología , Vectores Genéticos/genética , Vectores Genéticos/inmunología , Animales , Western Blotting , Electroforesis en Gel de Poliacrilamida , Citometría de Flujo , VIH-1/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Linfocitos T/inmunologíaRESUMEN
Egg drop syndrome (EDS) virus vaccines are routinely produced in embryonated duck eggs (Solyom et al., 1982). This procedure poses the risk of dissemination of pathogens, such as avian influenza virus, as the eggs used are not from specific pathogen free birds. To address this problem, the knob and part of the shaft domain of the fibre protein of the EDS virus (termed knob-s) were expressed in Escherichia coli and assessed as a subunit vaccine. A single vaccination with the recombinant protein induced the production of anti-EDS virus antibodies, as detected by haemagglutination inhibition, enzyme-linked immunosorbent assay and virus neutralization tests, for at least 20 weeks. A positive correlation was demonstrated between these three assays. A dose-response assessment showed that the vaccine was effective over the range of 2 to 64 microg protein per dose. Two vaccinations with the recombinant protein, administered before the onset of lay, induced high haemagglutination inhibition antibody titres, comparable with those induced by an inactivated whole-virus vaccine. The vaccine did not have any adverse effects on egg production, quality or weight. The present study has shown that two vaccinations with the recombinant knob-s protein elicited high neutralizing antibody titres that persisted for more than 50 weeks of lay.
Asunto(s)
Atadenovirus/inmunología , Pollos , Patos , Óvulo/virología , Vacunas Sintéticas/inmunología , Vacunas Virales/inmunología , Cultivo de Virus/veterinaria , Animales , Anticuerpos Antivirales/sangre , Clonación Molecular , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Pruebas de Inhibición de Hemaglutinación/veterinaria , Pruebas de Neutralización/veterinaria , Factores de Tiempo , Proteínas Virales/genética , Cultivo de Virus/métodosRESUMEN
Four 3-month-old Jersey calves and three 3-month-old Holstein calves were inoculated with cervid adenovirus and monitored for clinical signs until necropsied between 10 and 42 days postinoculation. The neonatal Jersey calves had received colostrum, and the Holstein calves were colostrum deprived. Preinoculation and postinoculation serum samples were tested for antibodies to the cervid adenovirus, bovine adenovirus type 6, bovine adenovirus type 7, and goat adenovirus type 1. Virus isolation was performed on kidney, nasal secretion, and/or lung homogenates in fetal white-tailed deer lung cells. Negatively stained preparations of feces from Jersey calves were examined weekly using an electron microscope, and weekly blood samples were collected for complete blood counts. Full necropsies were performed on all calves. A complete selection of tissues was evaluated for microscopic changes, and immunohistochemistry was performed on all tissues using a polyclonal antibody to deer adenovirus. No clinical signs were observed in the calves during the study period. Following inoculation, colostrum-deprived calves developed low antibody titers to deer adenovirus, while the Jersey calves that received colostrum did not. Calves that received colostrum had high antibody titers to bovine adenovirus type 7 and goat adenovirus type 1. No consistent gross or microscopic lesions were seen. Adenovirus was not observed in negatively stained preparations of feces. Immunohistochemistry results did not demonstrate virus in all tissues examined microscopically, and virus was not isolated from lungs, nasal secretions, and kidneys.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Atadenovirus/patogenicidad , Enfermedades de los Bovinos/virología , Bovinos/virología , Infecciones por Adenoviridae/inmunología , Infecciones por Adenoviridae/virología , Animales , Animales Recién Nacidos , Anticuerpos Antivirales/sangre , Atadenovirus/inmunología , Recuento de Células Sanguíneas/veterinaria , Bovinos/inmunología , Enfermedades de los Bovinos/inmunología , Calostro/inmunología , Ciervos , Heces/microbiología , Inmunohistoquímica/veterinaria , Masculino , Microscopía Electrónica/veterinaria , Pruebas de Neutralización/veterinariaRESUMEN
In current study, phagocytosis product (PP) of peripheral blood monocytes was detected among 920 dwarf chickens (460 per sex) at 20 wk of age, and based on discrepancies of PP, the flock was grouped (the highest group, the medium group, and the lowest group). Then serum hemagglutination inhibition antibody titers and subpopulations of T-lymphocytes of each group were examined after inoculations of avian influenza virus H5N2 inactivated vaccine (20 wk of age), avian influenza virus H9 inactivated vaccine (24 wk of age), and Newcastle disease virus-egg drop syndrome virus bigeminal inactivated vaccine (28 wk of age), respectively, to study the relationship between PP and immune response. To gain insight into effects of selection for PP on number of eggs, mean egg weight, fertilization rate, hatchability, and rate of healthy chicks, 9 (3 x 3) mating combinations were conducted. The results showed that (1) selection for higher PP in both sexes benefited to humoral immunity but not CD8(+) T-lymphocyte mediated immunity in dwarf chickens; (2) there were effects of selection for higher PP in hens on fertilization rate (P < 0.05), hatchability (P < 0.05), rate of healthy chicks (P < 0.05), and level of IgY antibody (P < 0.0001); however, hens' PP had no effects on number of eggs (P > or = 0.05) or egg weight (P > or = 0.05) and cocks' PP had no effect (P > or = 0.05) on any trait mentioned above. The results indicated that phagocytosis of peripheral blood monocytes might be an indicator of humoral immunity in dwarf chickens; furthermore, selection of hens with higher PP was not only beneficial to fertilization rate, but also benefited to hatchability and rate of healthy chicks in that the hens had stronger humoral immunity, which might contribute to maternal antibody in eggs.
Asunto(s)
Pollos/genética , Pollos/inmunología , Fagocitosis/genética , Fagocitosis/inmunología , Animales , Anticuerpos Antivirales/sangre , Formación de Anticuerpos/inmunología , Atadenovirus/inmunología , Huevos , Femenino , Citometría de Flujo/veterinaria , Pruebas de Inhibición de Hemaglutinación/veterinaria , Inmunización/métodos , Inmunización/veterinaria , Inmunoglobulinas/inmunología , Subtipo H2N2 del Virus de la Influenza A/inmunología , Subtipo H5N2 del Virus de la Influenza A/inmunología , Masculino , Virus de la Enfermedad de Newcastle/inmunología , Oviposición/inmunología , Selección Genética , Linfocitos T/inmunología , Vacunas de Productos Inactivados/inmunologíaRESUMEN
A simple, user-friendly, and rapid method to detect the presence of antibodies to egg drop syndrome 76 (EDS) virus in chicken sera based on an immunofiltration (flow-through) test was developed. Purified EDS virus antigen was coated onto nitrocellulose membranes housed in a plastic module with layers of absorbent filter pads underneath. Following addition of serum to be tested and washing, monoclonal antibodies or polyclonal serum to chicken immunoglobulin G (IgG) was used as a bridge antibody to mediate binding between EDS virus-specific IgG and protein A gold conjugate. The appearance of a pink dot indicated the presence of antibodies to EDS virus in the sample tested. The results could be obtained within 5-10 min. The developed immunofiltration test could detect antibodies in the sera of experimentally vaccinated chickens from 2 wk postvaccination. With field sera samples, this test was positive in samples having hemagglutination inhibition titers of 8 and above. This test has the potential to be used as a field-based kit to assess seroconversion in EDS-vaccinated flocks.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Anticuerpos Antivirales/sangre , Atadenovirus/inmunología , Pollos/inmunología , Inmunoensayo/veterinaria , Enfermedades de las Aves de Corral/inmunología , Infecciones por Adenoviridae/sangre , Infecciones por Adenoviridae/inmunología , Animales , Pollos/sangre , Pruebas de Inhibición de Hemaglutinación/veterinaria , Inmunoensayo/métodos , Enfermedades de las Aves de Corral/sangre , Enfermedades de las Aves de Corral/virología , Sensibilidad y EspecificidadRESUMEN
Adenoviruses have become a popular vehicle for gene transfer into animal and human cells. However, wide prevalence of preexisting immunity to human adenovirus (HAdV) and the promiscuous nature of the virus have made the use of nonhuman adenoviruses an attractive alternative. Moreover, readministration of viral vectors is often required to maintain therapeutic levels of transgene expression, resulting in vector-specific immune responses. Although a number of features of bovine adenovirus (BAdV)-3 make it attractive for use as a vector in human vaccination, BAdV-3 transduces nonbovine cells, including human cells, poorly. However, genetic modification of capsid proteins (e.g., fiber, pIX) has helped in increasing the utility of BAdV-3 as a vector for transducing nonbovine cells. Here, we will describe the methods used to construct recombinant BAdV-3 expressing chimeric fiber or chimeric pIX proteins.
Asunto(s)
Atadenovirus/genética , Proteínas de la Cápside/genética , Recombinación Genética , Animales , Atadenovirus/inmunología , Atadenovirus/aislamiento & purificación , Bovinos , Técnicas de Transferencia de Gen , Vectores Genéticos , Humanos , Plásmidos , Mapeo Restrictivo , Vacunas ViralesRESUMEN
To investigate the pathogens that racing pigeons in Taiwan are exposed to, a total of 3764 pigeons from 90 lofts were analysed by collection of blood samples in the period between October 2000 and September 2001. The haemagglutination inhibition (HI) test was performed to detect antibodies against Newcastle disease virus (NDV), type 2 avian paramyxovirus (APMV-2), and egg drop syndrome '76 virus (EDS-76V). The agar-gel precipitin (AGP) test was used to detect antibodies against fowl adenovirus (FAV), goose parvovirus (GPV), and avian reovirus (REO). The virus neutralisation (VN) test was applied to detect antibodies against the serotypes FAV-1 and FAV-8. A rapid serum agglutination test was applied for the detection of antibodies against Mycoplasma spp. Antibodies to several infectious agents were found, including NDV (43.3%), EDS-76V (19.2%), FAV (0.8%), REO (0.5%), APMV-2 (0.2%), Mycoplasma columbinum (10.3%), M. columborale (7.1%), M. synoviae (1.8%) and M. gallisepticum (1.3%). Antibodies against GPV, FAV-1, and FAV-8 were not detected in any serum sample. NDV seroprevalence was significantly higher in pigeons of more than one year of age than in pigeons younger than one year. ND or EDS-76 seroprevalence of pigeons vaccinated with ND vaccine or EDS-76 vaccine was significantly higher than that of pigeons that did not receive any vaccination.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Atadenovirus/inmunología , Infecciones por Avulavirus/veterinaria , Avulavirus/inmunología , Enfermedades de las Aves/epidemiología , Columbidae , Infecciones por Mycoplasma/veterinaria , Infecciones por Adenoviridae/epidemiología , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antivirales/sangre , Infecciones por Avulavirus/epidemiología , Enfermedades de las Aves/microbiología , Enfermedades de las Aves/virología , Mycoplasma/inmunología , Infecciones por Mycoplasma/epidemiología , Enfermedad de Newcastle/epidemiología , Virus de la Enfermedad de Newcastle/inmunología , Estudios Seroepidemiológicos , Taiwán/epidemiologíaRESUMEN
Mucosal sites are one of the main natural ports of entry into the body. Stimulation of a local response by antibodies as the systemic protection may enhance the efficacy of non-living vaccines, and allow for vaccination by subunit vaccines without the need for injection. Mucosal or skin vaccination necessitates a suitable adjuvant and carrier. Escherichia coli heat-labile enterotoxin (LT) and its B subunit (LTB) have been found to be effective adjuvants. The aim of this study was to efficiently produce and purify recombinant LTB (brLTB), and examine its adjuvant and carrier properties. The gene encoding LTB was cloned and expressed in E. coli, and the product was found to have a pentameric form with the ability to bind the cell receptor, GM1 ganglioside. A one-step method for efficient purification and concentration of brLTB was developed. Both oral and intramuscular vaccination with purified brLTB yielded high antibody titers, which detected the whole toxin. In an attempt to test its adjuvant characteristics, brLTB was mixed with either BSA or a recombinant protein (rKnob of egg drop syndrome adenovirus) and delivered intramuscularly, orally or transcutaneously. The addition of brLTB significantly elevated the antibody response in groups vaccinated orally and transcutaneously, but had no influence in injected groups. Vaccination with another recombinant protein, (viral protein 2 of infectious bursal disease virus) supplemented with brLTB did not elevate the antibody response, as compared to vaccination with the antigen alone. These results demonstrate that the addition of brLTB makes oral and transcutaneous vaccination with protein antigens possible.