Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus
Porcine epidemic diarrhea virus (PEDV) is a causative agent of a highly infectious disease with a high mortality rate, especially in newborn piglets in Asian countries resulting in serious economic loss. The development of a rapid, safe, effective and cost-efficient vaccine is crucial to protect pig...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-09-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.02152/full |
| _version_ | 1818200725747924992 |
|---|---|
| author | Thuong Thi Ho Thuong Thi Ho Giang Thu Nguyen Ngoc Bich Pham Ngoc Bich Pham Van Phan Le Thi Bich Ngoc Trinh Trang Huyen Vu Hoang Trong Phan Udo Conrad Ha Hoang Chu Ha Hoang Chu |
| author_facet | Thuong Thi Ho Thuong Thi Ho Giang Thu Nguyen Ngoc Bich Pham Ngoc Bich Pham Van Phan Le Thi Bich Ngoc Trinh Trang Huyen Vu Hoang Trong Phan Udo Conrad Ha Hoang Chu Ha Hoang Chu |
| author_sort | Thuong Thi Ho |
| collection | DOAJ |
| description | Porcine epidemic diarrhea virus (PEDV) is a causative agent of a highly infectious disease with a high mortality rate, especially in newborn piglets in Asian countries resulting in serious economic loss. The development of a rapid, safe, effective and cost-efficient vaccine is crucial to protect pigs against PEDV infection. The COE antigen is regarded to be a major target for subunit vaccine development against PEDV infection. The naturally assembled COE protein forms a homotrimeric structure. In the present study, we successfully produced a trimeric COE protein as a native structure by fusion with the C-terminal isoleucine zipper trimerization (GCN4pII) motif in Nicotiana benthamiana, with a high expression level shown via semi-quantified Western blots. Trimeric COE protein was purified via immobilized metal affinity chromatography (IMAC), and its trimeric structure was successfully demonstrated by a cross-linking reaction, and a native PAGE gel. A crude extract containing the COE trimer was used for evaluating immunogenicity in mice. After 1 and 2 booster immunizations, the crude extract containing trimeric COE elicited elevated PEDV-specific humoral responses, as demonstrated by ELISA and Western blot analyses. Notably, a virus-neutralizing antibody assay indicated that the neutralization activities of sera of mice vaccinated with the crude extract containing COE-GCN4pII were similar to those of mice vaccinated with a commercial vaccine. These results suggest that crude extract containing trimeric COE is a promising plant-based subunit vaccine candidate for PEDV prevention. |
| first_indexed | 2024-12-12T02:42:14Z |
| format | Article |
| id | doaj.art-98357ce16cb7471592915dc2e5d01659 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-12T02:42:14Z |
| publishDate | 2020-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-98357ce16cb7471592915dc2e5d016592022-12-22T00:41:08ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-09-011110.3389/fimmu.2020.02152552837Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea VirusThuong Thi Ho0Thuong Thi Ho1Giang Thu Nguyen2Ngoc Bich Pham3Ngoc Bich Pham4Van Phan Le5Thi Bich Ngoc Trinh6Trang Huyen Vu7Hoang Trong Phan8Udo Conrad9Ha Hoang Chu10Ha Hoang Chu11Graduate University of Science and Technology, Vietnam Academy of Science and Technology, Ha Noi, VietnamInstitute of Biotechnology, Vietnam Academy of Science and Technology, Ha Noi, VietnamInstitute of Biotechnology, Vietnam Academy of Science and Technology, Ha Noi, VietnamGraduate University of Science and Technology, Vietnam Academy of Science and Technology, Ha Noi, VietnamInstitute of Biotechnology, Vietnam Academy of Science and Technology, Ha Noi, VietnamVietnam National University of Agriculture, Ha Noi, VietnamVietnam National University of Agriculture, Ha Noi, VietnamInstitute of Biotechnology, Vietnam Academy of Science and Technology, Ha Noi, VietnamLeibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, GermanyLeibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, GermanyGraduate University of Science and Technology, Vietnam Academy of Science and Technology, Ha Noi, VietnamInstitute of Biotechnology, Vietnam Academy of Science and Technology, Ha Noi, VietnamPorcine epidemic diarrhea virus (PEDV) is a causative agent of a highly infectious disease with a high mortality rate, especially in newborn piglets in Asian countries resulting in serious economic loss. The development of a rapid, safe, effective and cost-efficient vaccine is crucial to protect pigs against PEDV infection. The COE antigen is regarded to be a major target for subunit vaccine development against PEDV infection. The naturally assembled COE protein forms a homotrimeric structure. In the present study, we successfully produced a trimeric COE protein as a native structure by fusion with the C-terminal isoleucine zipper trimerization (GCN4pII) motif in Nicotiana benthamiana, with a high expression level shown via semi-quantified Western blots. Trimeric COE protein was purified via immobilized metal affinity chromatography (IMAC), and its trimeric structure was successfully demonstrated by a cross-linking reaction, and a native PAGE gel. A crude extract containing the COE trimer was used for evaluating immunogenicity in mice. After 1 and 2 booster immunizations, the crude extract containing trimeric COE elicited elevated PEDV-specific humoral responses, as demonstrated by ELISA and Western blot analyses. Notably, a virus-neutralizing antibody assay indicated that the neutralization activities of sera of mice vaccinated with the crude extract containing COE-GCN4pII were similar to those of mice vaccinated with a commercial vaccine. These results suggest that crude extract containing trimeric COE is a promising plant-based subunit vaccine candidate for PEDV prevention.https://www.frontiersin.org/article/10.3389/fimmu.2020.02152/fulltrimeric COEPEDVneutralizing antibodyplant-based vaccinerecombinant protein |
| spellingShingle | Thuong Thi Ho Thuong Thi Ho Giang Thu Nguyen Ngoc Bich Pham Ngoc Bich Pham Van Phan Le Thi Bich Ngoc Trinh Trang Huyen Vu Hoang Trong Phan Udo Conrad Ha Hoang Chu Ha Hoang Chu Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus Frontiers in Immunology trimeric COE PEDV neutralizing antibody plant-based vaccine recombinant protein |
| title | Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus |
| title_full | Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus |
| title_fullStr | Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus |
| title_full_unstemmed | Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus |
| title_short | Plant-Derived Trimeric CO-26K-Equivalent Epitope Induced Neutralizing Antibodies Against Porcine Epidemic Diarrhea Virus |
| title_sort | plant derived trimeric co 26k equivalent epitope induced neutralizing antibodies against porcine epidemic diarrhea virus |
| topic | trimeric COE PEDV neutralizing antibody plant-based vaccine recombinant protein |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.02152/full |
| work_keys_str_mv | AT thuongthiho plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT thuongthiho plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT giangthunguyen plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT ngocbichpham plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT ngocbichpham plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT vanphanle plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT thibichngoctrinh plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT tranghuyenvu plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT hoangtrongphan plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT udoconrad plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT hahoangchu plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus AT hahoangchu plantderivedtrimericco26kequivalentepitopeinducedneutralizingantibodiesagainstporcineepidemicdiarrheavirus |