RESUMEN
Acinetobacter baumannii (A. Baumannii) is an emerging opportunistic pathogen responsible for hospital-acquired infections, and which now constitutes a sufficiently serious threat to public health to necessitate the development of an effective vaccine. In this study, a recombinant fused protein named OmpK/Omp22 and two individual proteins OmpK and Omp22 were obtained using recombinant expression and Ni-affinity purification. Groups of BALB/c mice were immunized with these proteins and challenged with a clinically isolated strain of A. baumannii. The bacterial load in the blood, pathological changes in the lung tissue and survival rates after challenge were evaluated. Mice immunized with OmpK/Omp22 fused protein provided significantly greater protection against A. baumannii challenge than those immunized with either of the two proteins individually. The results provide novel clues for future design of vaccines against A. baumannii.
Asunto(s)
Animales , Femenino , Infecciones por Acinetobacter , Patología , Acinetobacter baumannii , Genética , Alergia e Inmunología , Anticuerpos Antibacterianos , Sangre , Carga Bacteriana , Proteínas de la Membrana Bacteriana Externa , Genética , Alergia e Inmunología , Vacunas Bacterianas , Alergia e Inmunología , Modelos Animales de Enfermedad , Ratones Endogámicos BALB C , Neumonía Bacteriana , Patología , Proteínas Recombinantes de Fusión , Genética , Alergia e InmunologíaRESUMEN
OBJECTIVE:To study the characteristics of ceftazidime-resistant?-lactamase produced by Escherichia coli.METHODS:The types of2strains of drug fast?-lactamase produced by Escherichia coli were determined initially by K-B slip diffusion method and ampholine electrophoresis method;The plasmid was extracted by alkaline lysis and the PCR ampli?fication and sequencing were conducted;?-Lactamase was counter-extracted by saturated ammonium sulfate,filtrated by Sephadex G-75gel and purified by DE-52anion exchange chromatography;The molecular weight of which was determined by SDS-PAGE and the enzyme kinetics parameters of?-lactamas were determined by ultraviolet spectrophotometry.RE?SULTS:The2strains produced a super-broad spectrum?-lactamase(CTX-M-1V)with isoionic point at8.7and the molecular weight at29kDa,which can hydrolyze cefotaxim and aztreonam but imipenem and which was sensitive to sulbactam(IC 50 =94nmol/L)and tazobactam(IC 50 =5nmol/L).CONCLUSION:CTX-M-1V is a CTX-M type super-broad spectrum?-lactamase sensitive to suppressants.