A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice
IntroductionThe objective of this study is to construct a multi-epitope vaccine GILE containing B-cell and T-cell epitopes against Echinococcus Multilocularis (E. multilocularis) infection based on the dominant epitopes of E. multilocularis EMY162, LAP, and GLUT1.MethodsThe structure and hydrophobic...
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ftdoajarticles:oai:doaj.org/article:35cd972334874edc8005a14247611911 2023-05-15T15:18:23+02:00 A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice Pei Zhou Zhen Zhou Meiduo Huayu Lei Wang Lin Feng Yang Xiao Yao Dai Mingyuan Xin Feng Tang Runle Li 2023-01-01T00:00:00Z https://doi.org/10.3389/fimmu.2022.1091004 https://doaj.org/article/35cd972334874edc8005a14247611911 EN eng Frontiers Media S.A. https://www.frontiersin.org/articles/10.3389/fimmu.2022.1091004/full https://doaj.org/toc/1664-3224 1664-3224 doi:10.3389/fimmu.2022.1091004 https://doaj.org/article/35cd972334874edc8005a14247611911 Frontiers in Immunology, Vol 13 (2023) Echinococcus multilocularis multi-epitope vaccine prokaryotic expression immune bioinformatics Immunologic diseases. Allergy RC581-607 article 2023 ftdoajarticles https://doi.org/10.3389/fimmu.2022.1091004 2023-01-22T01:39:22Z IntroductionThe objective of this study is to construct a multi-epitope vaccine GILE containing B-cell and T-cell epitopes against Echinococcus Multilocularis (E. multilocularis) infection based on the dominant epitopes of E. multilocularis EMY162, LAP, and GLUT1.MethodsThe structure and hydrophobicity of GILE were predicted by SWISSMODEL, pyMOL, SOPMA and VMD, and its sequence was optimized by Optimum™ Codon. The GILE gene was inserted into pCzn1 and transformed into Escherichia coli Arctic express competent cells. IPTG was added to induce the expression of recombinant proteins. High-purity GILE recombinant protein was obtained by Ni-NTA Resin. BALB/c mice were immunized with GILE mixed with Freund’s adjuvant, and the antibody levels and dynamic changes in the serum were detected by ELISA. Lymphocyte proliferation was detected by MTS. The levels of IFN-g and IL-4 were detected by ELISpot and flow cytometry (FCM). T cells were detected by FCM. The growth of hepatic cysts was evaluated by Ultrasound and their weights were measured to evaluate the immune protective effect of GILE.ResultsThe SWISS-MODEL analysis showed that the optimal model was EMY162 95-104―LAP464-479―LAP495-510―LAP396-410―LAP504-518―EMY162112-126. The SOPMA results showed that there were Alpha helix (14.88%), Extended strand (26.25%), Beta turn (3.73%) and Random coil (45.82%) in the secondary structure of GILE. The restriction enzyme digestion and sequencing results suggested that the plasmid pCzn1-GILE was successfully constructed. The SDSPAGE results indicated that the recombinant protein was 44.68 KD. The ELISA results indicated that mice immunized with GILE showed higher levels of serum antibodies compared to the PBS group. The FCM and ELISpot results indicated that mice immunized with GILE secreted more IFN-g and IL-4. Immunization with GILE also led to a significant decrease in the maximum diameter and weight of cysts and stimulated the production of CD4+ and CD8+ T Cell.DiscussionA multi-epitope vaccine GILE with good immunogenicity and ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Frontiers in Immunology 13 |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Echinococcus multilocularis multi-epitope vaccine prokaryotic expression immune bioinformatics Immunologic diseases. Allergy RC581-607 |
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Echinococcus multilocularis multi-epitope vaccine prokaryotic expression immune bioinformatics Immunologic diseases. Allergy RC581-607 Pei Zhou Zhen Zhou Meiduo Huayu Lei Wang Lin Feng Yang Xiao Yao Dai Mingyuan Xin Feng Tang Runle Li A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
topic_facet |
Echinococcus multilocularis multi-epitope vaccine prokaryotic expression immune bioinformatics Immunologic diseases. Allergy RC581-607 |
description |
IntroductionThe objective of this study is to construct a multi-epitope vaccine GILE containing B-cell and T-cell epitopes against Echinococcus Multilocularis (E. multilocularis) infection based on the dominant epitopes of E. multilocularis EMY162, LAP, and GLUT1.MethodsThe structure and hydrophobicity of GILE were predicted by SWISSMODEL, pyMOL, SOPMA and VMD, and its sequence was optimized by Optimum™ Codon. The GILE gene was inserted into pCzn1 and transformed into Escherichia coli Arctic express competent cells. IPTG was added to induce the expression of recombinant proteins. High-purity GILE recombinant protein was obtained by Ni-NTA Resin. BALB/c mice were immunized with GILE mixed with Freund’s adjuvant, and the antibody levels and dynamic changes in the serum were detected by ELISA. Lymphocyte proliferation was detected by MTS. The levels of IFN-g and IL-4 were detected by ELISpot and flow cytometry (FCM). T cells were detected by FCM. The growth of hepatic cysts was evaluated by Ultrasound and their weights were measured to evaluate the immune protective effect of GILE.ResultsThe SWISS-MODEL analysis showed that the optimal model was EMY162 95-104―LAP464-479―LAP495-510―LAP396-410―LAP504-518―EMY162112-126. The SOPMA results showed that there were Alpha helix (14.88%), Extended strand (26.25%), Beta turn (3.73%) and Random coil (45.82%) in the secondary structure of GILE. The restriction enzyme digestion and sequencing results suggested that the plasmid pCzn1-GILE was successfully constructed. The SDSPAGE results indicated that the recombinant protein was 44.68 KD. The ELISA results indicated that mice immunized with GILE showed higher levels of serum antibodies compared to the PBS group. The FCM and ELISpot results indicated that mice immunized with GILE secreted more IFN-g and IL-4. Immunization with GILE also led to a significant decrease in the maximum diameter and weight of cysts and stimulated the production of CD4+ and CD8+ T Cell.DiscussionA multi-epitope vaccine GILE with good immunogenicity and ... |
format |
Article in Journal/Newspaper |
author |
Pei Zhou Zhen Zhou Meiduo Huayu Lei Wang Lin Feng Yang Xiao Yao Dai Mingyuan Xin Feng Tang Runle Li |
author_facet |
Pei Zhou Zhen Zhou Meiduo Huayu Lei Wang Lin Feng Yang Xiao Yao Dai Mingyuan Xin Feng Tang Runle Li |
author_sort |
Pei Zhou |
title |
A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
title_short |
A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
title_full |
A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
title_fullStr |
A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
title_full_unstemmed |
A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice |
title_sort |
multi-epitope vaccine gile against echinococcus multilocularis infection in mice |
publisher |
Frontiers Media S.A. |
publishDate |
2023 |
url |
https://doi.org/10.3389/fimmu.2022.1091004 https://doaj.org/article/35cd972334874edc8005a14247611911 |
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Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Frontiers in Immunology, Vol 13 (2023) |
op_relation |
https://www.frontiersin.org/articles/10.3389/fimmu.2022.1091004/full https://doaj.org/toc/1664-3224 1664-3224 doi:10.3389/fimmu.2022.1091004 https://doaj.org/article/35cd972334874edc8005a14247611911 |
op_doi |
https://doi.org/10.3389/fimmu.2022.1091004 |
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Frontiers in Immunology |
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13 |
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1766348572206301184 |