Development and Evaluation of a Peptide Amphiphiles based Multi-epitope Vaccine against Enterovirus 71
- 주제(키워드) Enterovirus 71 , Infectious disease , Epitope , Peptide amphiphiles , Vaccine , Immune response , Cytokine
- 발행기관 아주대학교
- 지도교수 진효언
- 발행년도 2020
- 학위수여년월 2020. 8
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000030119
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Hand-foot-and-mouth disease (HFMD) is an infectious disease that usually occurs in children under five years of age. Mainly caused by Coxsackievirus A16 (CA16) or Enterovirus 71 (EV71). Hand-foot-and-mouth disease (HFMD) caused by the CA16 infection has mostly mild symptoms. However, if the cause of EV71 appears, serious neurological complications such as meningitis and encephalitis may occur. But there is no vaccine and medicines for EV71 infection yet. Therefore, to prevent EV71, multi-epitope vaccine research based on peptide amphiphiles (PAs) was conducted. For research purposes, epitope was selected in capsid protein of EV71, and the form of conjugated fatty acids of different lengths was synthesized. The nanofiber type immune stimulator was formed through self-assembly of each composed PAs in aqueous condition. To evaluate the immunogenicity and efficacy for a PAs-based immune stimulator, constructed PAs were injection route of subcutaneous (s.c.) into Balb/c female mice. One week after the last injection, serum and spleen samples were collected. Through ELISA and the Western blot, it was confirmed antigen specific to the particles of EV71. In addition, cytokine reactions formed by stimulating splenocytes were measured (IFN-γ, IL-2, IL-10, IL-12 and IL-17). Stimulation index was confirmed through MTT assay after treating antigen in splenocyte. In conclusion, this study reported that the immune response induced by multi-epitope vaccine based on peptide amphiphiles induces improved immunity from peptide and single peptide-amphiphiles. Thus, peptide amphiphiles-based multi-epitope vaccines might be a new approach that could improve the low immunogenicity of conventional peptide vaccines.
more목차
1. Introduction １
1.1 Hand, Foot, and Mouth Diseases (HFMD) １
1.2 Enterovirus 71 １
1.3 Vaccine ３
1.4 Peptide amphiphiles ４
1.5 Multi-epitope vaccine ４
1.6 Aims of study ５
2. Materials and methods 7
2.1. Material 7
2.2. Reagents and apparatus 7
2.2.1. Reagents 7
2.2.2. Apparatus 7
2.3 Methods 8
2.3.1. Target e viral capsid protein election and cloning 8
2.3.2. Expression, purification and characterization of target protein 8
2.3.4. Preparation and construction peptide amphiphiles 10
2.3.5. Characterized by atomic force microscope 11
2.3.6. Animal study 11
2.3.7. Immune response assessment using ELISA 12
2.3.8. Immune response assessment using Western blot 13
2.3.9. Measurement antigen-specific proliferation 13
2.3.10. Analysis induce cytokine 14
2.3.11. Statistical analysis 14
3. Results and Discussion 15
3.1 Previous study 15
3.2 Recombinant virus protein was expression in E.coli and specific antigenicity 18
3.3 Epitope selection 20
3.4 Epitope peptide amphiphiles preparation 22
3.5 Self-assembly to Peptide amphiphiles and characterization 24
3.6 Comparative splenic size and morphology 26
3.7 Evaluation of humoral immune response by Enzyme-linked immunosorbent assay (ELISA) 30
3.8 Evaluation of specific-IgG detection by Western blot 33
3.9 proliferation response of the splenocytes. 37
3.10 Analysis of the production of cytokine from splenocyte. 39
4. Conclusion 42
References 43
-국문요약- 47
