An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli
Objective(s): Enterotoxigenic Escherichia coli (ETEC) is known as the most common bacterial causes of diarrheal diseases related to morbidity and mortality. Heat-labile enterotoxin (LT) is a part of major virulence factors in ETEC pathogenesis. Antigen entrapment into nanoparticles (NPs) can protect...
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| Format: | Article |
| Language: | English |
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Mashhad University of Medical Sciences
2018-05-01
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| Series: | Iranian Journal of Basic Medical Sciences |
| Subjects: | |
| Online Access: | http://ijbms.mums.ac.ir/article_10545_abc4c98e982432fb8391d05f1a9faaf5.pdf |
| _version_ | 1818522206890622976 |
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| author | Emad Kordbacheh Shahram Nazarian Davood Sadeghi Abbas Hajizade |
| author_facet | Emad Kordbacheh Shahram Nazarian Davood Sadeghi Abbas Hajizade |
| author_sort | Emad Kordbacheh |
| collection | DOAJ |
| description | Objective(s): Enterotoxigenic Escherichia coli (ETEC) is known as the most common bacterial causes of diarrheal diseases related to morbidity and mortality. Heat-labile enterotoxin (LT) is a part of major virulence factors in ETEC pathogenesis. Antigen entrapment into nanoparticles (NPs) can protect them and enhance their immunogenicity.Materials and Methods: In the present study, recombinant LTB protein was expressed in E. coli BL21 (DE3) and purified by an Ni-NTA agarose column. The protein was entrapped in PLGA polymer by the double emulsion method. NPs were characterized physicochemically and the protein release from the NPs was evaluated. ELISA assay was performed for investigation of raised antibody against the recombinant protein in mice. The anti-toxicity and anti-adherence attributes of the immune sera against ETEC were also evaluated.Results: It showed the successful cloning of a 313 bp DNA fragment encoding LTB protein in the pET28a vector. Over-expression in BL21 (DE3) led to the formation of corresponding 15.5 kDa protein bands in the SDS-PAGE gel. Western blotting by using anti-CTX confirmed the purified LTB. Protein-entrapped NPs had a spherical shape with the size of 238 nm mean diameter and 85% entrapment efficiency. Immunological analyses showed the production of a high titer of specific IgG antibody in immunized animals. The neutralizing antibody in the sera of immunized animals was approved by GM1 binding and Ileal loop assays.Conclusion: The results indicate the efficacy of the entrapped LTB protein as an effective immunogen which induces the humoral responses. |
| first_indexed | 2024-12-11T05:30:14Z |
| format | Article |
| id | doaj.art-302aff78ae644a5bbd014ad2bd6b21ca |
| institution | Directory Open Access Journal |
| issn | 2008-3866 2008-3874 |
| language | English |
| last_indexed | 2024-12-11T05:30:14Z |
| publishDate | 2018-05-01 |
| publisher | Mashhad University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Basic Medical Sciences |
| spelling | doaj.art-302aff78ae644a5bbd014ad2bd6b21ca2022-12-22T01:19:27ZengMashhad University of Medical SciencesIranian Journal of Basic Medical Sciences2008-38662008-38742018-05-0121551752410.22038/ijbms.2018.27017.660910545An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coliEmad Kordbacheh0Shahram Nazarian1Davood Sadeghi2Abbas Hajizade3Imam Hossein University, Faculty of Science, Department of Biology, Tehran, IranImam Hossein University, Faculty of Science, Department of Biology, Tehran, IranImam Hossein University, Faculty of Science, Department of Biology, Tehran, IranImam Hossein University, Faculty of Science, Department of Biology, Tehran, IranObjective(s): Enterotoxigenic Escherichia coli (ETEC) is known as the most common bacterial causes of diarrheal diseases related to morbidity and mortality. Heat-labile enterotoxin (LT) is a part of major virulence factors in ETEC pathogenesis. Antigen entrapment into nanoparticles (NPs) can protect them and enhance their immunogenicity.Materials and Methods: In the present study, recombinant LTB protein was expressed in E. coli BL21 (DE3) and purified by an Ni-NTA agarose column. The protein was entrapped in PLGA polymer by the double emulsion method. NPs were characterized physicochemically and the protein release from the NPs was evaluated. ELISA assay was performed for investigation of raised antibody against the recombinant protein in mice. The anti-toxicity and anti-adherence attributes of the immune sera against ETEC were also evaluated.Results: It showed the successful cloning of a 313 bp DNA fragment encoding LTB protein in the pET28a vector. Over-expression in BL21 (DE3) led to the formation of corresponding 15.5 kDa protein bands in the SDS-PAGE gel. Western blotting by using anti-CTX confirmed the purified LTB. Protein-entrapped NPs had a spherical shape with the size of 238 nm mean diameter and 85% entrapment efficiency. Immunological analyses showed the production of a high titer of specific IgG antibody in immunized animals. The neutralizing antibody in the sera of immunized animals was approved by GM1 binding and Ileal loop assays.Conclusion: The results indicate the efficacy of the entrapped LTB protein as an effective immunogen which induces the humoral responses.http://ijbms.mums.ac.ir/article_10545_abc4c98e982432fb8391d05f1a9faaf5.pdfEnterotoxigenic Escherichia coliHeat-labile enterotoxin ImmunizationNanoparticlePLGA |
| spellingShingle | Emad Kordbacheh Shahram Nazarian Davood Sadeghi Abbas Hajizade An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli Iranian Journal of Basic Medical Sciences Enterotoxigenic Escherichia coli Heat-labile enterotoxin Immunization Nanoparticle PLGA |
| title | An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli |
| title_full | An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli |
| title_fullStr | An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli |
| title_full_unstemmed | An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli |
| title_short | An LTB-entrapped protein in PLGA nanoparticles preserves against enterotoxin of enterotoxigenic Escherichia coli |
| title_sort | ltb entrapped protein in plga nanoparticles preserves against enterotoxin of enterotoxigenic escherichia coli |
| topic | Enterotoxigenic Escherichia coli Heat-labile enterotoxin Immunization Nanoparticle PLGA |
| url | http://ijbms.mums.ac.ir/article_10545_abc4c98e982432fb8391d05f1a9faaf5.pdf |
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