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1.
Vet Immunol Immunopathol ; 190: 65-72, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28778325

RESUMEN

We have recently reported that grass pollen allergoids conjugated with nonoxidized mannan of Saccharomyces cerevisae using glutaraldehyde results in a novel hypoallergenic mannan-allergen complex with improved properties for allergen vaccination. Using this approach, human dendritic cells show a better allergen uptake and cytokine profile production (higher IL-10/IL-4 ratio) for therapeutic purposes. Here we aim to address whether a similar approach can be extended to dogs using canine dendritic cells. Six healthy Spanish Greyhound dogs were used as blood donors to obtain canine dendritic cells (DC) derived from peripheral blood monocytes. Allergens from Dermatophagoides farinae mite were polymerized and conjugated with nonoxidized mannan. Nuclear magnetic resonance (NMR), gel electrophoresis (SDS-PAGE), immunoblotting and IgE-ELISA inhibition studies were conducted to evaluate the main characteristics of the allergoid obtained. Mannan-allergen conjugate and controls were assayed in vitro for canine DC uptake and production of IL-4 and IL-10. The results indicate that the conjugation of D. farinae allergens with nonoxidized mannan was feasible using glutaraldehyde. The resulting product was a polymerized structure showing a high molecular weight as detected by NMR and SDS-PAGE analysis. The mannan-allergen conjugate was hypoallergenic with a reduced reactivity with specific dog IgE. An increase in both allergen uptake and IL-10/IL-4 ratio was obtained when canine DCs were incubated with the mannan-allergen conjugate, as compared with the control allergen preparations (unmodified D. farinae allergens and oxidized mannan-allergen conjugate). We conclude that hypoallergenic D. farinae allergens coupled to nonoxidized mannan is a novel allergen preparation suitable for canine allergy immunotherapy targeting dendritic cells.


Asunto(s)
Antígenos Dermatofagoides/inmunología , Células Dendríticas/inmunología , Enfermedades de los Perros/terapia , Hipersensibilidad/veterinaria , Inmunoterapia/veterinaria , Mananos/inmunología , Saccharomyces cerevisiae/inmunología , Animales , Enfermedades de los Perros/inmunología , Perros , Electroforesis en Gel de Poliacrilamida/veterinaria , Hipersensibilidad/inmunología , Hipersensibilidad/terapia , Immunoblotting/veterinaria , Inmunoterapia/métodos , Espectroscopía de Resonancia Magnética
2.
J Rheumatol ; 32(3): 405-9, 2005 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-15742429

RESUMEN

OBJECTIVE: Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology characterized by cartilage and bone destruction. The main genetic determinant to RA, the shared epitope, maps to the HLA-DR locus, although this is not the only risk factor. The osteopontin (OPN) gene, with pleiotropic functions in inflammatory and immune responses, has been implicated in the pathogenesis of RA. We studied the association of polymorphisms in the OPN gene and predisposition to RA. METHODS: Analysis was performed in a case-control study with 263 patients and 478 controls. Four single nucleotide polymorphisms (SNP), 327T/C, 795C/T, 1128A/G, and 1284A/C, of the OPN gene were genotyped by primer-specific amplification in the presence of SYBR Green. RESULTS: Distorted transmission of these polymorphisms was studied in 58 RA trios and 61 affected sibling pairs. These SNP demonstrated strong linkage disequilibrium. No statistically significant association was observed (80% power to exclude a genotypic relative risk of 1.49 at the 5% significance level, with minor allele frequencies of 28%). This lack of association with RA was found after stratification for the shared epitope as well. CONCLUSION: Our data suggest that, unlike the reported effect of the OPN SNP conferring predisposition to common diseases such as multiple sclerosis or systemic lupus erythematosus, these OPN gene polymorphisms do not contribute to RA susceptibility in the Spanish population we studied.


Asunto(s)
Artritis Reumatoide/genética , Polimorfismo Genético , Sialoglicoproteínas/genética , Artritis Reumatoide/inmunología , Estudios de Casos y Controles , Epítopos , Predisposición Genética a la Enfermedad , Antígenos HLA-DR/genética , Antígenos HLA-DR/inmunología , Haplotipos , Humanos , Osteopontina , Factores de Riesgo , España
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