RESUMEN
Porcine rotavirus (PoRV) and porcine epidemic diarrhea virus (PEDV) usually co-infect pigs in modern large-scale piggery, which both can cause severe diarrhea in newborn piglets and lead to significant economic losses to the pig industry. The VP7 protein is the main coat protein of PoRV, and the S protein is the main structural protein of PEDV, which are capable of inducing neutralizing antibodies in vivo. In this study, a DNA vaccine pPI-2.EGFP.VP7.S co-expressing VP7 protein of PoRV and S protein of PEDV was constructed. Six 8-week-old mice were immunized with the recombinant plasmid pPI-2.EGFP.VP7.S. The high humoral immune responses (virus specific antibody) and cellular immune responses (IFN-γ, IL-4, and spleen lymphocyte proliferation) were evaluated. The immune effect through intramuscular injection increased with plasmid dose when compared with subcutaneous injection. The immune-enhancing effect of IFN-α adjuvant was excellent compared with pig spleen transfer factor and IL-12 adjuvant. These results demonstrated that pPI-2.EGFP.VP7.S possess the immunological functions of the VP7 proteins of PoRV and S proteins of PEDV, indicating that pPI-2.EGFP.VP7.S is a candidate vaccine for porcine rotaviral infection (PoR) and porcine epidemic diarrhea (PED).
Asunto(s)
Antígenos Virales/inmunología , Proteínas de la Cápside/inmunología , Infecciones por Coronavirus/veterinaria , Plásmidos/inmunología , Infecciones por Rotavirus/veterinaria , Rotavirus/inmunología , Enfermedades de los Porcinos/prevención & control , Proteínas Virales de Fusión/inmunología , Vacunas Virales/inmunología , Animales , Antígenos Virales/administración & dosificación , Antígenos Virales/genética , Proteínas de la Cápside/administración & dosificación , Proteínas de la Cápside/genética , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/prevención & control , Infecciones por Coronavirus/virología , ADN Recombinante/administración & dosificación , ADN Recombinante/genética , ADN Recombinante/inmunología , Evaluación Preclínica de Medicamentos , Ratones , Plásmidos/administración & dosificación , Plásmidos/genética , Virus de la Diarrea Epidémica Porcina/genética , Virus de la Diarrea Epidémica Porcina/inmunología , Rotavirus/genética , Infecciones por Rotavirus/inmunología , Infecciones por Rotavirus/prevención & control , Infecciones por Rotavirus/virología , Porcinos , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/virología , Proteínas Virales de Fusión/administración & dosificación , Proteínas Virales de Fusión/genética , Vacunas Virales/administración & dosificación , Vacunas Virales/genéticaRESUMEN
BACKGROUND: Previous studies have established that mycobacterial infections ameliorate allergic inflammation. However, a non-infectious approach that controls allergic responses might represent a safer and more promising strategy. The 60-65 kDa heat shock protein (Hsp) family is endowed with anti-inflammatory properties, but it is still unclear whether and how single mycobacterial Hsp control allergic disorders. OBJECTIVE: Therefore, in this study we determined whether the administration of Mycobacterial leprae Hsp65 expressed by recombinant a DNA plasmid could attenuate a previously established allergic response. METHODS: We used an experimental model of airway allergic inflammation to test the effects of immunotherapy with DNA encoding Hsp65. Allergic mice, previously sensitized and challenged with ovalbumin, were treated with tree intramuscular doses of recombinant DNA encoding Hsp65. After treatment, mice received a second allergen challenge and the allergic response was measured. RESULTS: We found that immunotherapy attenuated eosinophilia, pulmonary inflammation, Th2 cytokine and mucus production. Moreover, we showed that the inhibition of allergic response is dependent on IL-10 production. Both Hsp65 and allergen-specific IL-10-producing cells contributed to this effect. Cells transferred from DNA-immunized mice to allergic mice migrated to allergic sites and down-modulated the Th2 response. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings clearly show that immunotherapy with DNA encoding Hsp65 can attenuate an established Th2 allergic inflammation through an IL-10-dependent mechanism; moreover, the migration of allergen- and Hsp65-specific cells to the allergic sites exerts a fundamental role. This work represents a novel contribution to the understanding of immune regulation by Hsp65 in allergic diseases.
Asunto(s)
Proteínas Bacterianas , Chaperonina 60 , ADN Recombinante/administración & dosificación , Inmunoterapia/métodos , Interleucina-10/metabolismo , Hipersensibilidad Respiratoria/terapia , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Chaperonina 60/genética , Chaperonina 60/inmunología , ADN Recombinante/inmunología , Modelos Animales de Enfermedad , Femenino , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Interleucina-10/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Mycobacterium leprae/inmunología , Hipersensibilidad Respiratoria/inmunología , Resultado del TratamientoRESUMEN
By using retroviral insertional mutagenesis in zebrafish, we have identified a recessive lethal mutation in the not really started (nrs) gene. The nrs mutation disrupts a gene located in linkage group 3 that is highly homologous to the spinster gene identified in Drosophila and to spinster orthologs identified in mammals. In flies, spinster encodes a membrane protein involved in lysosomal metabolism and programmed cell death in the central nervous system and in the ovary. In nrs mutant fish embryos, we detect an opaque substance in the posterior yolk cell extension at approximately 1 day after fertilization. This material progressively accumulates and by 48 hr after fertilization fills the entire yolk. By day 3 of embryogenesis, mutant embryos are severely reduced in size compared with their wild-type siblings and they die a few hours later. By in situ hybridization, we show that the nrs mRNA is expressed in the yolk cell at the time the mutant phenotype becomes apparent. In wild-type embryos, nrs message is present maternally and zygotically throughout embryogenesis and is also detected in adult animals. In nrs homozygous mutant embryos, nrs transcripts are undetectable at the time the phenotype becomes apparent, indicating that the retroviral insertion has most likely abolished expression of the nrs gene. Finally, the nrs phenotype can be partially rescued by microinjection of nrs encoding DNA. These results suggest that the nrs mutation affects an essential gene encoding a putative membrane-bound protein expressed specifically in the yolk cell during zebrafish embryogenesis.
Asunto(s)
Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Proteínas del Huevo/genética , Genes , Proteínas de la Membrana/genética , Proteínas de Pez Cebra/genética , Pez Cebra/genética , Secuencia de Aminoácidos , Animales , Proteínas de Ciclo Celular , Mapeo Cromosómico , ADN Recombinante/administración & dosificación , ADN Recombinante/genética , Proteínas de Drosophila/fisiología , Proteínas del Huevo/fisiología , Etiquetas de Secuencia Expresada , Femenino , Regulación del Desarrollo de la Expresión Génica , Marcación de Gen , Genes Letales , Ligamiento Genético , Humanos , Hibridación in Situ , Proteínas de la Membrana/deficiencia , Proteínas de la Membrana/fisiología , Proteínas de Transporte de Membrana , Ratones , Microinyecciones , Proteínas Asociadas a Microtúbulos , Datos de Secuencia Molecular , Mutagénesis Insercional , Fosfoproteínas , ARN Mensajero/genética , Retroviridae/genética , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Especificidad de la Especie , Pez Cebra/embriología , Proteínas de Pez Cebra/deficiencia , Proteínas de Pez Cebra/fisiologíaRESUMEN
Reactogenicity was measured after applying the Heberbiovac-HB recombinant vaccine against hepatitis B virus to three groups of children aged 6-9 years. The vaccine was derived from yeast cells, administered at doses of 10, 5, and 2.5 g, with a schedule of 0, 1 and 6 months. The overall observed symptomatology was low (12.2%) in the three groups with 10.7%, 13.5%, and 12.3% for 10, 5, and 2.5 g, respectively. The predominant symptoms and signs were febricula (7.0%), local pain (3.1%), and erythema (1.2%). No significant differences were found when making a comparison between groups and sexes. An acceptable reactogenicity of the immunogen was confirmed, thus its use is recommended for the protection against hepatitis B virus.
Asunto(s)
Vacunas contra Hepatitis B/efectos adversos , Vacunas de ADN/efectos adversos , Niño , Cuba , ADN Recombinante/administración & dosificación , ADN Recombinante/efectos adversos , Relación Dosis-Respuesta Inmunológica , Hepatitis B/prevención & control , Vacunas contra Hepatitis B/administración & dosificación , Humanos , Inyecciones Intramusculares , Factores de Tiempo , Vacunas de ADN/administración & dosificaciónRESUMEN
The development of recombinant vector vaccines will be guided by nearly two centuries of research in vaccinology and immunology. Experimental vector vaccines may be of viral, bacterial or genetic composition and their acceptability will depend on safety, efficacy, and practicality as seen by the user, the developer, and the licensing authority. Recombinant vector vaccines will need to compete with alternative vaccine approaches and may find special use for non-propagable agents, immunization in early life, fertility control, and emerging infectious agents. Needs of the developing nations that include low cost, temperature stability, and ease of administration may be met, in part at least, by the vector vaccine approach.