Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study
Abstract Brucellosis is regarded as one of the world’s most severe zoonotic diseases. This study aimed to investigate the possibility of using recombinant Lactococcus lactis (L. lactis) as a live vector to produce recombinant Brucella abortus (B. abortus) Omp10. The gene sequences were obtained from...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
SpringerOpen
2024-01-01
|
| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-023-01638-4 |
| _version_ | 1797362864604839936 |
|---|---|
| author | Mahsa Kazemi-Roudsari Abbas Doosti Mohammad-Saeid Jami |
| author_facet | Mahsa Kazemi-Roudsari Abbas Doosti Mohammad-Saeid Jami |
| author_sort | Mahsa Kazemi-Roudsari |
| collection | DOAJ |
| description | Abstract Brucellosis is regarded as one of the world’s most severe zoonotic diseases. This study aimed to investigate the possibility of using recombinant Lactococcus lactis (L. lactis) as a live vector to produce recombinant Brucella abortus (B. abortus) Omp10. The gene sequences were obtained from GenBank. The proteins’ immunogenicity was assessed using Vaxijen. After confirming the cloning of the Omp10 gene in the pNZ8148 vector by enzymatic digestion and PCR, transformation into L. lactis was done. SDS-PAGE and western blot methods evaluated omp10 protein expression. Mice received oral recombinant L. lactis vaccines. IgG antibodies against Omp10 were tested using ELISA. Real-time PCR and ELISA were used to analyze cytokine responses. Survival rate and histopathological changes were evaluated after the challenge. Omp10 was chosen for its 1.5524 antigenicity score. Enzymatic digestion and PCR identified a 381-bp gene fragment. A 10 kDa band indicated the success of L. lactis transformation. Mice administered the L. lactis-pNZ8148-Omp10-Usp45 vaccination 14 days after priming showed significantly higher Omp10-specific total IgG and IgG1 (P < 0.001) than the PBS control group. The mice who received the L. lactis-pNZ8148-Omp10-Usp45 and IRBA vaccines had significantly elevated levels of IFN-γ, TNFα, IL-4, and IL-10 in samples collected on days 14 and 28 (P < 0.001). Inflammatory response, morphological damage, alveolar edema, and lymphocyte infiltration were reduced in the target group. A recombinant L. lactis expressing the Omp10 protein was constructed as an oral Lactococcus-based vaccine and compared to live attenuated vaccines for future brucellosis investigations. |
| first_indexed | 2024-03-08T16:13:33Z |
| format | Article |
| id | doaj.art-a33af7467a6b440b961e33c350455f47 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-03-08T16:13:33Z |
| publishDate | 2024-01-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-a33af7467a6b440b961e33c350455f472024-01-07T12:48:23ZengSpringerOpenAMB Express2191-08552024-01-0114111410.1186/s13568-023-01638-4Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo studyMahsa Kazemi-Roudsari0Abbas Doosti1Mohammad-Saeid Jami2Department of Biology, Shahrekord Branch, Islamic Azad UniversityBiotechnology Research Center, Shahrekord Branch, Islamic Azad UniversityDepartment of Biology, Shahrekord Branch, Islamic Azad UniversityAbstract Brucellosis is regarded as one of the world’s most severe zoonotic diseases. This study aimed to investigate the possibility of using recombinant Lactococcus lactis (L. lactis) as a live vector to produce recombinant Brucella abortus (B. abortus) Omp10. The gene sequences were obtained from GenBank. The proteins’ immunogenicity was assessed using Vaxijen. After confirming the cloning of the Omp10 gene in the pNZ8148 vector by enzymatic digestion and PCR, transformation into L. lactis was done. SDS-PAGE and western blot methods evaluated omp10 protein expression. Mice received oral recombinant L. lactis vaccines. IgG antibodies against Omp10 were tested using ELISA. Real-time PCR and ELISA were used to analyze cytokine responses. Survival rate and histopathological changes were evaluated after the challenge. Omp10 was chosen for its 1.5524 antigenicity score. Enzymatic digestion and PCR identified a 381-bp gene fragment. A 10 kDa band indicated the success of L. lactis transformation. Mice administered the L. lactis-pNZ8148-Omp10-Usp45 vaccination 14 days after priming showed significantly higher Omp10-specific total IgG and IgG1 (P < 0.001) than the PBS control group. The mice who received the L. lactis-pNZ8148-Omp10-Usp45 and IRBA vaccines had significantly elevated levels of IFN-γ, TNFα, IL-4, and IL-10 in samples collected on days 14 and 28 (P < 0.001). Inflammatory response, morphological damage, alveolar edema, and lymphocyte infiltration were reduced in the target group. A recombinant L. lactis expressing the Omp10 protein was constructed as an oral Lactococcus-based vaccine and compared to live attenuated vaccines for future brucellosis investigations.https://doi.org/10.1186/s13568-023-01638-4BrucellosisLactococcus lactisOmp10Oral vaccine |
| spellingShingle | Mahsa Kazemi-Roudsari Abbas Doosti Mohammad-Saeid Jami Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study AMB Express Brucellosis Lactococcus lactis Omp10 Oral vaccine |
| title | Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study |
| title_full | Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study |
| title_fullStr | Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study |
| title_full_unstemmed | Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study |
| title_short | Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study |
| title_sort | design of an oral vaccine using lactococcus lactis against brucellosis an in vitro and in vivo study |
| topic | Brucellosis Lactococcus lactis Omp10 Oral vaccine |
| url | https://doi.org/10.1186/s13568-023-01638-4 |
| work_keys_str_mv | AT mahsakazemiroudsari designofanoralvaccineusinglactococcuslactisagainstbrucellosisaninvitroandinvivostudy AT abbasdoosti designofanoralvaccineusinglactococcuslactisagainstbrucellosisaninvitroandinvivostudy AT mohammadsaeidjami designofanoralvaccineusinglactococcuslactisagainstbrucellosisaninvitroandinvivostudy |