RESUMEN
In order to develop a specific assay for the measurement of goat IL-18 level, two stable hybridoma cell lines were established which secreted IgG1 monoclonal antibodies (mAbs) against goat IL-18. Specific binding of two mAbs named 2E8 and 4C4 to recombinant goat IL-18 expressed in Escherichia coli was demonstrated in an ELISA and Western blotting. Results also showed that mAbs 2E8 and 4C4 bound to distinct epitopes in the ELISA additivity test. These two mAbs were applied in IFA analysis for the detection of goat IL-18 expressed in 293FT cells and in the sandwich ELISA for the measurement of goat IL-18 levels in LPS-stimulated PBMC. Results from this study demonstrated that mAbs against goat IL-18 recognize bovine and human IL-18 and could be used to measure IL-18 levels in different inflammations or immune responses in future studies.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Cabras/inmunología , Interleucina-18/análisis , Interleucina-18/inmunología , Animales , Secuencia de Bases , Bovinos , Cartilla de ADN/genética , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Células HEK293 , Humanos , Técnicas In Vitro , Interleucina-18/genética , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Lipopolisacáridos/farmacología , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/análisis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Especificidad de la EspecieRESUMEN
Avian hepatitis E virus (HEV) has been identified in chickens; however, only 4 complete or near-complete genomic sequences have been reported. We found that the near-complete genomic sequence of avian HEV in chickens from China shared the highest identity (98.3%) with avian HEV from Europe and belonged to avian HEV genotype 3.