<i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes
(1) Background and Purpose: Ebola virus (EBOV) is the causative agent of Ebola virus disease (EVD), which causes extremely high mortality and widespread epidemics. The only glycoprotein (GP) on the surface of EBOV particles is the key to mediating viral invasion into host cells. DNA vaccines for EBO...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-10-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/11/10/1620 |
| _version_ | 1797572066500673536 |
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| author | Junqi Zhang Baozeng Sun Wenyang Shen Zhenjie Wang Yang Liu Yubo Sun Jiaxing Zhang Ruibo Liu Yongkai Wang Tianyuan Bai Zilu Ma Cheng Luo Xupeng Qiao Xiyang Zhang Shuya Yang Yuanjie Sun Dongbo Jiang Kun Yang |
| author_facet | Junqi Zhang Baozeng Sun Wenyang Shen Zhenjie Wang Yang Liu Yubo Sun Jiaxing Zhang Ruibo Liu Yongkai Wang Tianyuan Bai Zilu Ma Cheng Luo Xupeng Qiao Xiyang Zhang Shuya Yang Yuanjie Sun Dongbo Jiang Kun Yang |
| author_sort | Junqi Zhang |
| collection | DOAJ |
| description | (1) Background and Purpose: Ebola virus (EBOV) is the causative agent of Ebola virus disease (EVD), which causes extremely high mortality and widespread epidemics. The only glycoprotein (GP) on the surface of EBOV particles is the key to mediating viral invasion into host cells. DNA vaccines for EBOV are in development, but their effectiveness is unclear. The lack of immune characteristics resides in antigenic MHC class II reactivity. (2) Methods: We selected MHC-II molecules from four human leukocyte antigen II (HLA-II) superfamilies with 98% population coverage and eight mouse H2-I alleles. IEDB, NetMHCIIpan, SYFPEITHI, and Rankpep were used to screen MHC-II-restricted epitopes with high affinity for EBOV GP. Further immunogenicity and conservation analyses were performed using VaxiJen and BLASTp, respectively. EpiDock was used to simulate molecular docking. Cluster analysis and binding affinity analysis of EBOV GP epitopes and selected MHC-II molecules were performed using data from NetMHCIIpan. The selective GP epitopes were verified by the enzyme-linked immunospot (ELISpot) assay using splenocytes of BALB/c (H2d), C3H, and C57 mice after DNA vaccine pVAX-GP<sub>EBO</sub> immunization. Subsequently, BALB/c mice were immunized with Protein-GP<sub>EBO</sub>, plasmid pVAX-GP<sub>EBO</sub>, and pVAX-LAMP/GP<sub>EBO</sub>, which encoded EBOV GP. The dominant epitopes of BALB/c (H-2-I-AdEd genotype) mice were verified by the enzyme-linked immunospot (ELISpot) assay. It is also used to evaluate and explore the advantages of pVAX-LAMP/GP<sub>EBO</sub> and the reasons behind them. (3) Results: Thirty-one HLA-II-restricted and 68 H2-I-restricted selective epitopes were confirmed to have high affinity, immunogenicity, and conservation. Nineteen selective epitopes have cross-species reactivity with good performance in MHC-II molecular docking. The ELISpot results showed that pVAX-GP<sub>EBO</sub> could induce a cellular immune response to the synthesized selective peptides. The better immunoprotection of the DNA vaccines pVAX-LAMP/GP<sub>EBO</sub> coincides with the enhancement of the MHC class II response. (4) Conclusions: Promising MHC-II-restricted candidate epitopes of EBOV GP were identified in humans and mice, which is of great significance for the development and evaluation of Ebola vaccines. |
| first_indexed | 2024-03-10T20:49:29Z |
| format | Article |
| id | doaj.art-0b60db9d70c644ce8974a0976212b490 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T20:49:29Z |
| publishDate | 2023-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-0b60db9d70c644ce8974a0976212b4902023-11-19T18:25:29ZengMDPI AGVaccines2076-393X2023-10-011110162010.3390/vaccines11101620<i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted EpitopesJunqi Zhang0Baozeng Sun1Wenyang Shen2Zhenjie Wang3Yang Liu4Yubo Sun5Jiaxing Zhang6Ruibo Liu7Yongkai Wang8Tianyuan Bai9Zilu Ma10Cheng Luo11Xupeng Qiao12Xiyang Zhang13Shuya Yang14Yuanjie Sun15Dongbo Jiang16Kun Yang17Department of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaInstitute of AIDS Prevention and Control, Shaanxi Provincial Center for Disease Control and Prevention, Xi’an 710054, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, ChinaDepartment of Immunology, Basic Medicine School, Air-Force Medical University (The Fourth Military Medical University), Xi’an 710032, China(1) Background and Purpose: Ebola virus (EBOV) is the causative agent of Ebola virus disease (EVD), which causes extremely high mortality and widespread epidemics. The only glycoprotein (GP) on the surface of EBOV particles is the key to mediating viral invasion into host cells. DNA vaccines for EBOV are in development, but their effectiveness is unclear. The lack of immune characteristics resides in antigenic MHC class II reactivity. (2) Methods: We selected MHC-II molecules from four human leukocyte antigen II (HLA-II) superfamilies with 98% population coverage and eight mouse H2-I alleles. IEDB, NetMHCIIpan, SYFPEITHI, and Rankpep were used to screen MHC-II-restricted epitopes with high affinity for EBOV GP. Further immunogenicity and conservation analyses were performed using VaxiJen and BLASTp, respectively. EpiDock was used to simulate molecular docking. Cluster analysis and binding affinity analysis of EBOV GP epitopes and selected MHC-II molecules were performed using data from NetMHCIIpan. The selective GP epitopes were verified by the enzyme-linked immunospot (ELISpot) assay using splenocytes of BALB/c (H2d), C3H, and C57 mice after DNA vaccine pVAX-GP<sub>EBO</sub> immunization. Subsequently, BALB/c mice were immunized with Protein-GP<sub>EBO</sub>, plasmid pVAX-GP<sub>EBO</sub>, and pVAX-LAMP/GP<sub>EBO</sub>, which encoded EBOV GP. The dominant epitopes of BALB/c (H-2-I-AdEd genotype) mice were verified by the enzyme-linked immunospot (ELISpot) assay. It is also used to evaluate and explore the advantages of pVAX-LAMP/GP<sub>EBO</sub> and the reasons behind them. (3) Results: Thirty-one HLA-II-restricted and 68 H2-I-restricted selective epitopes were confirmed to have high affinity, immunogenicity, and conservation. Nineteen selective epitopes have cross-species reactivity with good performance in MHC-II molecular docking. The ELISpot results showed that pVAX-GP<sub>EBO</sub> could induce a cellular immune response to the synthesized selective peptides. The better immunoprotection of the DNA vaccines pVAX-LAMP/GP<sub>EBO</sub> coincides with the enhancement of the MHC class II response. (4) Conclusions: Promising MHC-II-restricted candidate epitopes of EBOV GP were identified in humans and mice, which is of great significance for the development and evaluation of Ebola vaccines.https://www.mdpi.com/2076-393X/11/10/1620Ebola virus glycoprotein (EBOV GP)pan-MHC-II epitopeimmunoreactivityin silico analysisDNA vaccineenzyme-linked immunospot (ELISpot) assay |
| spellingShingle | Junqi Zhang Baozeng Sun Wenyang Shen Zhenjie Wang Yang Liu Yubo Sun Jiaxing Zhang Ruibo Liu Yongkai Wang Tianyuan Bai Zilu Ma Cheng Luo Xupeng Qiao Xiyang Zhang Shuya Yang Yuanjie Sun Dongbo Jiang Kun Yang <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes Vaccines Ebola virus glycoprotein (EBOV GP) pan-MHC-II epitope immunoreactivity in silico analysis DNA vaccine enzyme-linked immunospot (ELISpot) assay |
| title | <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes |
| title_full | <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes |
| title_fullStr | <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes |
| title_full_unstemmed | <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes |
| title_short | <i>In Silico</i> Analyses, Experimental Verification and Application in DNA Vaccines of Ebolavirus GP-Derived pan-MHC-II-Restricted Epitopes |
| title_sort | i in silico i analyses experimental verification and application in dna vaccines of ebolavirus gp derived pan mhc ii restricted epitopes |
| topic | Ebola virus glycoprotein (EBOV GP) pan-MHC-II epitope immunoreactivity in silico analysis DNA vaccine enzyme-linked immunospot (ELISpot) assay |
| url | https://www.mdpi.com/2076-393X/11/10/1620 |
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