RESUMEN
In our previous study, an ompP2 mutant of a Haemophilus parasuis SC096 strain showed sensitivity to serum bactericidal activity. In this study, we inactivated two gal genes, galU and galE, and these mutants were found to be serum sensitive to porcine sera. Furthermore, the galE mutant exhibited greater sensitivity than the galU mutant in serum sensitivity assays. Biofilm formation ability was also investigated. The galU mutant is unable to form biofilms, while more biofilm mass was produced by the galE mutant compared with SC096. Lack of expression of GalU protein by the galU mutant increased its tendency to autoagglutinate. The results indicated that the galU plays a role in autoagglutination and biofilm formation, while galE may affect the biofilm production indirectly. Both genes are significant for serum resistance in the H. parasuis SC096 strain.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Haemophilus parasuis/fisiología , UDPglucosa 4-Epimerasa/genética , UTP-Glucosa-1-Fosfato Uridililtransferasa/genética , Animales , Actividad Bactericida de la Sangre , Galactosemias , Haemophilus parasuis/enzimología , Haemophilus parasuis/genética , Haemophilus parasuis/inmunología , Mutación , Determinación de Anticuerpos Séricos Bactericidas , Porcinos , UDPglucosa 4-Epimerasa/inmunología , UTP-Glucosa-1-Fosfato Uridililtransferasa/inmunologíaRESUMEN
Schistosomiasis is an important global public health problem, as millions of people are at risk of acquiring this infection. An ideal method for sustainable control of schistosomiasis is using a vaccine alone or in combination with drugs. In the present study, we cloned the SjGALE gene and generated the expression product in E. coli. The expression level of SjGALE during different developmental stages of S. japonicum was evaluated by real-time RT-PCR and western blotting. Immunolocalization indicated that the protein was mainly located on the tegument of the parasite. Infection of rSjGALE-immunized mice demonstrated a 34% and 49% reduction of the mean worm burden and liver egg burden, respectively, in two independent experiments, indicating immune protection. The liver egg count from each female adult worm was significantly reduced by 63% in the two trials. The cytokine profile and IgG isotype analysis demonstrated the induction of a Th1 immune profile in response to immunization with this protein, further suggesting protection against infection. In conclusion, these findings indicated that SjGALE is a potential vaccine against S. japonicum.
Asunto(s)
Schistosoma japonicum/anatomía & histología , Schistosoma japonicum/enzimología , Esquistosomiasis Japónica/inmunología , UDPglucosa 4-Epimerasa/inmunología , UDPglucosa 4-Epimerasa/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos Antihelmínticos/inmunología , Clonación Molecular , ADN Complementario/genética , Femenino , Adyuvante de Freund/inmunología , Inmunidad Celular , Inmunidad Humoral , Inmunización , Estadios del Ciclo de Vida , Masculino , Ratones , Datos de Secuencia Molecular , Transporte de Proteínas , Schistosoma japonicum/crecimiento & desarrollo , Schistosoma japonicum/patogenicidad , Transcripción Genética , UDPglucosa 4-Epimerasa/química , UDPglucosa 4-Epimerasa/genéticaRESUMEN
Ganglioside-like structures in CAMPYLOBACTER JEJUNI ( C. JEJUNI) lipooligosaccharide (LOS) can induce antiganglioside antibodies, which might cause nerve damage. In this study, we injected the following three antisera directly into the sciatic nerve of guinea pigs, to investigate the role of anti-glycolipids antibody in inducing neural injury: (i) the wild strain antiserum, a mixture of the sera obtained from the guinea pigs immunized with C. JEJUNI wild-type strain (HS:19) that had a high titer anti-GM1 IgG antibody (range: 800-6,400; median: 2,400) and a high titer anti-LOS IgG antibody; (ii) the GALE mutant antiserum, a mixture of the sera obtained from the guinea pigs immunized with the GALE mutant strain that had only a high titer anti-LOS IgG antibody but no anti-GM1 antibody; and (iii) the control antiserum, a mixture of the sera obtained from the guinea pigs immunized with Freund's complete adjuvant alone which had no anti-GM1 or anti-LOS IgG antibody. Pathological examinations showed that the wild strain C. JEJUNI antiserum produced axonal degeneration in sciatic nerves. Demyelination was rare, and no inflammatory cells were present. The pathological features are consistent with those seen in human patients with axonal GBS. No such changes were observed in nerves injected with the GALE mutant antiserum. The experiment showed that passive transfer of serum containing high titer GM1 antibody caused axonal degeneration of peripheral nerves. The result, which reproduced our previous findings in an active immunization study, therefore further confirmed the critical role of the anti-glycolipid antibody in the induction of neuropathy.