RESUMEN

Edwardsiella tarda is a pathogen with a broad host range that infects both animals and humans. Resistance to phagocytic killing may be involved in the pathogenicity of this bacterium. Here we show that intracellular replication of E. tarda in murine macrophages is dependent on the type III secretion system and induces an anti-apoptotic effect by up-regulating anti-apoptotic NF-kappaB target genes. The wild-type strain replicates within the phagosomal membrane of macrophages; whereas the type III mutant does not. Microarray analysis shows the mRNA expression level of NF-kappaB target genes (e.g. pro-inflammatory cytokines and anti-apoptotic genes) in macrophages infected with the wild-type strain were up-regulated compared to macrophages infected with the type III mutant. Up-regulation of Bcl2a1a, Bcl2a1b, cIAP-2, and TRAF1 genes induced expression of anti-apoptotic proteins to protect macrophages from apoptosis induced by staurosporine. Further, this protection was inhibited by adding kamebakaurin, an inhibitor of NF-kappaB activation and was confirmed using an NF-kappaB reporter gene assay. Up-regulation of anti-apoptotic NF-kappaB target genes is responsible for the anti-apoptotic activity of E. tarda and is required for intracellular replication in murine macrophages.

Asunto(s)

Apoptosis/fisiología , Edwardsiella tarda/crecimiento & desarrollo , Infecciones por Enterobacteriaceae/microbiología , Macrófagos/microbiología , FN-kappa B/genética , Estaurosporina/farmacología , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Western Blotting , Línea Celular , ADN Bacteriano/genética , Diterpenos/farmacología , Edwardsiella tarda/genética , Edwardsiella tarda/metabolismo , Edwardsiella tarda/patogenicidad , Regulación Bacteriana de la Expresión Génica , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/fisiología , Ratones , Microscopía Electrónica de Transmisión , Antígenos de Histocompatibilidad Menor , Mutagénesis Insercional , FN-kappa B/antagonistas & inhibidores , FN-kappa B/biosíntesis , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Proteínas Proto-Oncogénicas c-bcl-2/genética , Regulación hacia Arriba