Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection
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....
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Hindawi Limited
2014-01-01
|
Series: | BioMed Research International |
Online Access: | http://dx.doi.org/10.1155/2014/421486 |
id |
doaj-39b1c6d413c54f2591c637ff747e62cd |
---|---|
record_format |
Article |
spelling |
doaj-39b1c6d413c54f2591c637ff747e62cd2020-11-25T00:44:08ZengHindawi LimitedBioMed Research International2314-61332314-61412014-01-01201410.1155/2014/421486421486Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis InfectionNoelia Olivera0Celina E. Castuma1Daniela Hozbor2María E. Gaillard3Martín Rumbo4Ricardo M. Gómez5Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT-La Plata, CONICET-UNLP, Calle 49 y 115, 1900 La Plata, ArgentinaInstituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT-La Plata, CONICET-UNLP, Calle 49 y 115, 1900 La Plata, ArgentinaInstituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT-La Plata, CONICET-UNLP, Calle 49 y 115, 1900 La Plata, ArgentinaInstituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT-La Plata, CONICET-UNLP, Calle 49 y 115, 1900 La Plata, ArgentinaLISIN, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Calle 48 y 115, 1900 La Plata, ArgentinaInstituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT-La Plata, CONICET-UNLP, Calle 49 y 115, 1900 La Plata, ArgentinaThis 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.http://dx.doi.org/10.1155/2014/421486 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Noelia Olivera Celina E. Castuma Daniela Hozbor María E. Gaillard Martín Rumbo Ricardo M. Gómez |
spellingShingle |
Noelia Olivera Celina E. Castuma Daniela Hozbor María E. Gaillard Martín Rumbo Ricardo M. Gómez Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection BioMed Research International |
author_facet |
Noelia Olivera Celina E. Castuma Daniela Hozbor María E. Gaillard Martín Rumbo Ricardo M. Gómez |
author_sort |
Noelia Olivera |
title |
Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection |
title_short |
Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection |
title_full |
Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection |
title_fullStr |
Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection |
title_full_unstemmed |
Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection |
title_sort |
immunization with the recombinant cholera toxin b fused to fimbria 2 protein protects against bordetella pertussis infection |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2014-01-01 |
description |
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. |
url |
http://dx.doi.org/10.1155/2014/421486 |
work_keys_str_mv |
AT noeliaolivera immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection AT celinaecastuma immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection AT danielahozbor immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection AT mariaegaillard immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection AT martinrumbo immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection AT ricardomgomez immunizationwiththerecombinantcholeratoxinbfusedtofimbria2proteinprotectsagainstbordetellapertussisinfection |
_version_ |
1725276166986661888 |