RESUMO
Aquatic animals are frequently threated by bacterial pathogens. The most economic and efficient protection against bacterial infection are through vaccine immunization. The various serotypes of the pathogens, such as Vibrios, hurdle the development of the vaccines, especially polyvalent vaccines. Here, we demonstrate that recombinant bacterial ghost is a good candidate for multivalent vaccine. By expressing PhiX174 gene E alone or co-expressing the gene E with two genes encoding outer membrane proteins (VP1667 and VP2369) in V. parahaemolyticus, we generated the recombinant V. parahaemolyticus ghosts VPG and rVPGs respectively. Fish immunized with either VPG or rVPG showed increased survival against the infection by either V. parahaemolyticus or V. alginolyticus, with a better protective effect by immunization with rVPG. Our furthermore studies show that rVPG stimulates stronger innate immune responses by increasing the expression of tnfα, il1ß, il6, il8 and il10 as well as that of c3b, lyz, and tlr5, the key players linking the innate and adaptive immune responses upon microbial stimulation. In summary, VPG and rVPG can protect zebrafish against the infection from at least two Vibrio species, suggesting its potential value for further aquaculture vaccines development.
Assuntos
Antibacterianos/farmacologia , Vacinas Bacterianas/farmacologia , Doenças dos Peixes/prevenção & controle , Vibrioses/veterinária , Vibrio parahaemolyticus/imunologia , Peixe-Zebra , Animais , Antibacterianos/metabolismo , Vacinas Bacterianas/genética , Vacinas Bacterianas/metabolismo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/prevenção & controleRESUMO
Vibrio cholerae, the causative agent of severe watery diarrheal disease cholera, requires production of a number of virulence factors during infection which results from the activity of a cascading system of regulatory factors by sensing to different environmental signals. TcpP, a membrane-localized transcription activator in V. cholerae, activates virulence factors production by responding to human host signals. To better characterize the transmembrane helix in regard to its roles on TcpP positive effectors sensitivity, site-directed mutagenesis was performed to identify specific mutations in this region which could enhance TcpP transcription activity in the absence of stimuli, like bile salts. We found that TcpP L152A constitutively forms homodimer and activates toxT expression in the absence of bile salts. However, being active, TcpP L152A needs to form disulfide bonds between the cysteine residues in the periplasmic domain of TcpP. We also found that TcpP L152A showed a competitive advantage in the infant mouse colonization model by coadministrating the bile salt-sequestering resin cholestyramine. All these results demonstrate that the transmembrane helix of TcpP plays an important role in regulating TcpP transcription activity in response to its positive effectors.