AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus
Japanese encephalitis virus (JEV) is an enveloped icosahedral capsid virus with a prime neutralizing epitope present in E protein domain III (EDIII). E dimers are rearranged into a five-fold symmetry of icosahedrons. Cholera toxin B (CTB) and heat-labile enterotoxin B (LTB) of AB<sub>5</sub...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-06-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-6651/15/7/425 |
| _version_ | 1797587349483290624 |
|---|---|
| author | Jina Ahn Ji Eun Yu Hanna Kim Jemin Sung Gyoonhee Han Myung Hyun Sohn Baik-Lin Seong |
| author_facet | Jina Ahn Ji Eun Yu Hanna Kim Jemin Sung Gyoonhee Han Myung Hyun Sohn Baik-Lin Seong |
| author_sort | Jina Ahn |
| collection | DOAJ |
| description | Japanese encephalitis virus (JEV) is an enveloped icosahedral capsid virus with a prime neutralizing epitope present in E protein domain III (EDIII). E dimers are rearranged into a five-fold symmetry of icosahedrons. Cholera toxin B (CTB) and heat-labile enterotoxin B (LTB) of AB<sub>5</sub>-type toxin was used as the structural scaffold for emulating the pentameric axis of EDIII. We produced homo-pentameric EDIII through the genetic fusion of LTB or CTB in <i>E. coli</i> without recourse to additional refolding steps. Harnessing an RNA-mediated chaperone further enhanced the soluble expression and pentameric assembly of the chimeric antigen. The pentameric assembly was validated by size exclusion chromatography (SEC), non-reduced gel analysis, and a GM1 binding assay. CTB/LTB−EDIII chimeric antigen triggered high neutralizing antibodies against the JEV Nakayama strain after immunization in mice. Altogether, our proof-of-principle study creating a JEV-protective antigen via fusion with an AB<sub>5</sub>-type toxin as both a pentameric scaffold and a built-in adjuvant posits the bacterially produced recombinant chimeric antigen as a cost-effective alternative to conventional inactivated vaccines against JEV. |
| first_indexed | 2024-03-11T00:35:19Z |
| format | Article |
| id | doaj.art-83458f20a44245b4b01d356cad491e4f |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-11T00:35:19Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-83458f20a44245b4b01d356cad491e4f2023-11-18T21:38:00ZengMDPI AGToxins2072-66512023-06-0115742510.3390/toxins15070425AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis VirusJina Ahn0Ji Eun Yu1Hanna Kim2Jemin Sung3Gyoonhee Han4Myung Hyun Sohn5Baik-Lin Seong6The Interdisciplinary Graduate Program in Integrative Biotechnology & Translational Medicine, Yonsei University, Incheon 21983, Republic of KoreaDepartment of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03708, Republic of KoreaDepartment of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03708, Republic of KoreaDepartment of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03708, Republic of KoreaDepartment of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03708, Republic of KoreaDepartment of Pediatrics, College of Medicine, Yonsei University, Seoul 03722, Republic of KoreaDepartment of Microbiology, College of Medicine, Yonsei University, Seoul 03722, Republic of KoreaJapanese encephalitis virus (JEV) is an enveloped icosahedral capsid virus with a prime neutralizing epitope present in E protein domain III (EDIII). E dimers are rearranged into a five-fold symmetry of icosahedrons. Cholera toxin B (CTB) and heat-labile enterotoxin B (LTB) of AB<sub>5</sub>-type toxin was used as the structural scaffold for emulating the pentameric axis of EDIII. We produced homo-pentameric EDIII through the genetic fusion of LTB or CTB in <i>E. coli</i> without recourse to additional refolding steps. Harnessing an RNA-mediated chaperone further enhanced the soluble expression and pentameric assembly of the chimeric antigen. The pentameric assembly was validated by size exclusion chromatography (SEC), non-reduced gel analysis, and a GM1 binding assay. CTB/LTB−EDIII chimeric antigen triggered high neutralizing antibodies against the JEV Nakayama strain after immunization in mice. Altogether, our proof-of-principle study creating a JEV-protective antigen via fusion with an AB<sub>5</sub>-type toxin as both a pentameric scaffold and a built-in adjuvant posits the bacterially produced recombinant chimeric antigen as a cost-effective alternative to conventional inactivated vaccines against JEV.https://www.mdpi.com/2072-6651/15/7/425chapernaJapanese encephalitis virusprotein foldingrecombinant vaccine |
| spellingShingle | Jina Ahn Ji Eun Yu Hanna Kim Jemin Sung Gyoonhee Han Myung Hyun Sohn Baik-Lin Seong AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus Toxins chaperna Japanese encephalitis virus protein folding recombinant vaccine |
| title | AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus |
| title_full | AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus |
| title_fullStr | AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus |
| title_full_unstemmed | AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus |
| title_short | AB<sub>5</sub>-Type Toxin as a Pentameric Scaffold in Recombinant Vaccines against the Japanese Encephalitis Virus |
| title_sort | ab sub 5 sub type toxin as a pentameric scaffold in recombinant vaccines against the japanese encephalitis virus |
| topic | chaperna Japanese encephalitis virus protein folding recombinant vaccine |
| url | https://www.mdpi.com/2072-6651/15/7/425 |
| work_keys_str_mv | AT jinaahn absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT jieunyu absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT hannakim absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT jeminsung absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT gyoonheehan absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT myunghyunsohn absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus AT baiklinseong absub5subtypetoxinasapentamericscaffoldinrecombinantvaccinesagainstthejapaneseencephalitisvirus |