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Biomed Res Int ; 2014: 421486, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24982881

RESUMEN

This study examined the immunogenic properties of the fusion protein fimbria 2 of Bordetella pertussis (Fim2)-cholera toxin B subunit (CTB) in the intranasal murine model of infection. To this end B. pertussis Fim2 coding sequence was cloned downstream of the cholera toxin B subunit coding sequence. The expression and assembly of the fusion protein into pentameric structures (CTB-Fim2) were evaluated by SDS-PAGE and monosialotetrahexosylgaglioside (GM1-ganglioside) enzyme-linked immunosorbent assay (ELISA). To evaluate the protective capacity of CTB-Fim2, an intraperitoneal or intranasal mouse immunization schedule was performed with 50 µg of CTB-Fim2. Recombinant (rFim2) or purified (BpFim2) Fim2, CTB, and phosphate-buffered saline (PBS) were used as controls. The results showed that mice immunized with BpFim2 or CTB-Fim2 intraperitoneally or intranasally presented a significant reduction in bacterial lung counts compared to control groups (P < 0.01 or P < 0.001 , resp.). Moreover, intranasal immunization with CTB-Fim2 induced significant levels of Fim2-specific IgG in serum and bronchoalveolar lavage (BAL) and Fim2-specific IgA in BAL. Analysis of IgG isotypes and cytokines mRNA levels showed that CTB-Fim2 results in a mixed Th1/Th2 (T-helper) response. The data presented here provide support for CTB-Fim2 as a promising recombinant antigen against Bordetella pertussis infection.


Asunto(s)
Antígenos Bacterianos/inmunología , Bordetella pertussis/inmunología , Toxina del Cólera/inmunología , Proteínas Fimbrias/inmunología , Inmunización , Proteínas Recombinantes de Fusión/inmunología , Factores de Virulencia de Bordetella/inmunología , Tos Ferina/microbiología , Tos Ferina/prevención & control , Animales , Líquido del Lavado Bronquioalveolar , Citocinas/genética , Citocinas/metabolismo , Femenino , Inmunidad Humoral , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Pulmón/metabolismo , Pulmón/microbiología , Pulmón/patología , Ratones Endogámicos BALB C , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes de Fusión/aislamiento & purificación , Tos Ferina/sangre , Tos Ferina/inmunología
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