{"title":"利用免疫信息学方法探索黄热病病毒的结构抗原,以设计多表位亚单位候选疫苗。","authors":"Kiran Sura, Himanshi Rohilla, Dev Kumar, Ritu Jakhar, Vaishali Ahlawat, Deepshikha Kaushik, Mehak Dangi, Anil Kumar Chhillar","doi":"10.1186/s43141-023-00621-7","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Yellow fever is a mosquito-borne viral hemorrhagic disease transmitted by several species of virus-infected mosquitoes endemic to tropical regions of Central and South America and Africa. Earlier in the twentieth century, mass vaccination integrated with mosquito control was implemented to eradicate the yellow fever virus. However, regular outbreaks occur in these regions which pose a threat to travelers and residents of Africa and South America. There is no specific antiviral therapy, but there can be an effective peptide-based vaccine candidate to combat infection caused by the virus. Therefore, the study aims to design a multi-epitope-based subunit vaccine (MESV) construct against the yellow fever virus to reduce the time and cost using reverse vaccinology (RV) approach.</p><p><strong>Methods: </strong>Yellow fever virus contains 10,233 nucleotides that encode for 10 proteins (C, prM, E, NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) including 3 structural and 7 non-structural proteins. Structural proteins-precursor membrane protein (prM) and envelope protein (E)-were taken as a target for B cell and T cell epitope screening. Further, various immunoinformatics approaches were employed to FASTA sequences of structural proteins to retrieve B cell and T cell epitopes. MESV was constructed from these epitopes based on allergenicity, antigenicity and immunogenicity, toxicity, conservancy, and population coverage followed by structure prediction. The efficacy of the MESV construct to bind with human TLR-3, TLR-4, and TLR-8 were evaluated using molecular docking and simulation studies. Finally, in-silico cloning of vaccine construct was performed withpBR322 Escherichia coli expression system using codon optimization.</p><p><strong>Results: </strong>Predicted epitopes evaluated and selected for MESV construction were found stable, non-allergenic, highly antigenic, and global population coverage of 68.03% according to in-silico analysis. However, this can be further tested in in-vitro and in-vivo investigations. Epitopes were sequentially merged to construct a MESV consisting of 393 amino acids using adjuvant and linkers. Molecular docking and simulation studies revealed stable and high-affinity interactions. Furthermore, in-silico immune response graphs showed effective immune response generation. Finally, higher CAI value ensured high gene expression of vaccine in the host cell.</p><p><strong>Conclusion: </strong>The designed MESV construct in the present in-silico study can be effective in generating an immune response against the yellow fever virus. Therefore, to prevent yellow fever, it can be an effective vaccine candidate. However, further downstream, in-vitro study is required.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":null,"pages":null},"PeriodicalIF":3.6000,"publicationDate":"2023-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10697922/pdf/","citationCount":"0","resultStr":"{\"title\":\"Exploring structural antigens of yellow fever virus to design multi-epitope subunit vaccine candidate by utilizing an immuno-informatics approach.\",\"authors\":\"Kiran Sura, Himanshi Rohilla, Dev Kumar, Ritu Jakhar, Vaishali Ahlawat, Deepshikha Kaushik, Mehak Dangi, Anil Kumar Chhillar\",\"doi\":\"10.1186/s43141-023-00621-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Yellow fever is a mosquito-borne viral hemorrhagic disease transmitted by several species of virus-infected mosquitoes endemic to tropical regions of Central and South America and Africa. Earlier in the twentieth century, mass vaccination integrated with mosquito control was implemented to eradicate the yellow fever virus. However, regular outbreaks occur in these regions which pose a threat to travelers and residents of Africa and South America. There is no specific antiviral therapy, but there can be an effective peptide-based vaccine candidate to combat infection caused by the virus. Therefore, the study aims to design a multi-epitope-based subunit vaccine (MESV) construct against the yellow fever virus to reduce the time and cost using reverse vaccinology (RV) approach.</p><p><strong>Methods: </strong>Yellow fever virus contains 10,233 nucleotides that encode for 10 proteins (C, prM, E, NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) including 3 structural and 7 non-structural proteins. Structural proteins-precursor membrane protein (prM) and envelope protein (E)-were taken as a target for B cell and T cell epitope screening. Further, various immunoinformatics approaches were employed to FASTA sequences of structural proteins to retrieve B cell and T cell epitopes. MESV was constructed from these epitopes based on allergenicity, antigenicity and immunogenicity, toxicity, conservancy, and population coverage followed by structure prediction. The efficacy of the MESV construct to bind with human TLR-3, TLR-4, and TLR-8 were evaluated using molecular docking and simulation studies. Finally, in-silico cloning of vaccine construct was performed withpBR322 Escherichia coli expression system using codon optimization.</p><p><strong>Results: </strong>Predicted epitopes evaluated and selected for MESV construction were found stable, non-allergenic, highly antigenic, and global population coverage of 68.03% according to in-silico analysis. However, this can be further tested in in-vitro and in-vivo investigations. Epitopes were sequentially merged to construct a MESV consisting of 393 amino acids using adjuvant and linkers. Molecular docking and simulation studies revealed stable and high-affinity interactions. Furthermore, in-silico immune response graphs showed effective immune response generation. Finally, higher CAI value ensured high gene expression of vaccine in the host cell.</p><p><strong>Conclusion: </strong>The designed MESV construct in the present in-silico study can be effective in generating an immune response against the yellow fever virus. Therefore, to prevent yellow fever, it can be an effective vaccine candidate. 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引用次数: 0
摘要
背景:黄热病是一种蚊媒病毒性出血性疾病,由中美洲、南美洲和非洲热带地区特有的几种受病毒感染的蚊子传播。二十世纪初,为根除黄热病病毒,采取了大规模疫苗接种和蚊虫控制相结合的措施。然而,这些地区经常爆发黄热病,对非洲和南美洲的旅行者和居民构成威胁。目前还没有特效的抗病毒疗法,但有一种有效的肽基候选疫苗可用于抗病毒感染。因此,本研究旨在利用逆向疫苗学(RV)方法设计一种针对黄热病病毒的多肽亚单位疫苗(MESV)构建体,以减少时间和成本:黄热病病毒含有10,233个核苷酸,编码10种蛋白(C、prM、E、NS1、NS2A、NS2B、NS3、NS4A、NS4B和NS5),包括3种结构蛋白和7种非结构蛋白。结构蛋白--前体膜蛋白(prM)和包膜蛋白(E)--被作为 B 细胞和 T 细胞表位筛选的目标。此外,还对结构蛋白的 FASTA 序列采用了各种免疫信息学方法,以检索 B 细胞和 T 细胞表位。根据这些表位的过敏性、抗原性和免疫原性、毒性、保守性和种群覆盖率构建 MESV,然后进行结构预测。利用分子对接和模拟研究评估了 MESV 构建物与人类 TLR-3、TLR-4 和 TLR-8 结合的功效。最后,利用密码子优化技术,使用pBR322大肠杆菌表达系统对疫苗构建体进行了体内克隆:结果:经评估和筛选用于构建 MESV 的预测表位具有稳定性、非过敏性、高抗原性,并且根据内嵌分析,全球群体覆盖率为 68.03%。不过,这还需要在体外和体内研究中进一步检验。利用佐剂和连接体将表位依次合并,构建了由 393 个氨基酸组成的 MESV。分子对接和模拟研究显示了稳定的高亲和性相互作用。此外,海量免疫反应图显示了有效的免疫反应生成。最后,较高的 CAI 值确保了疫苗在宿主细胞中的高基因表达:本研究中设计的 MESV 构建物能有效产生针对黄热病病毒的免疫反应。因此,它可以成为预防黄热病的有效候选疫苗。不过,还需要进一步的下游体外研究。
Exploring structural antigens of yellow fever virus to design multi-epitope subunit vaccine candidate by utilizing an immuno-informatics approach.
Background: Yellow fever is a mosquito-borne viral hemorrhagic disease transmitted by several species of virus-infected mosquitoes endemic to tropical regions of Central and South America and Africa. Earlier in the twentieth century, mass vaccination integrated with mosquito control was implemented to eradicate the yellow fever virus. However, regular outbreaks occur in these regions which pose a threat to travelers and residents of Africa and South America. There is no specific antiviral therapy, but there can be an effective peptide-based vaccine candidate to combat infection caused by the virus. Therefore, the study aims to design a multi-epitope-based subunit vaccine (MESV) construct against the yellow fever virus to reduce the time and cost using reverse vaccinology (RV) approach.
Methods: Yellow fever virus contains 10,233 nucleotides that encode for 10 proteins (C, prM, E, NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) including 3 structural and 7 non-structural proteins. Structural proteins-precursor membrane protein (prM) and envelope protein (E)-were taken as a target for B cell and T cell epitope screening. Further, various immunoinformatics approaches were employed to FASTA sequences of structural proteins to retrieve B cell and T cell epitopes. MESV was constructed from these epitopes based on allergenicity, antigenicity and immunogenicity, toxicity, conservancy, and population coverage followed by structure prediction. The efficacy of the MESV construct to bind with human TLR-3, TLR-4, and TLR-8 were evaluated using molecular docking and simulation studies. Finally, in-silico cloning of vaccine construct was performed withpBR322 Escherichia coli expression system using codon optimization.
Results: Predicted epitopes evaluated and selected for MESV construction were found stable, non-allergenic, highly antigenic, and global population coverage of 68.03% according to in-silico analysis. However, this can be further tested in in-vitro and in-vivo investigations. Epitopes were sequentially merged to construct a MESV consisting of 393 amino acids using adjuvant and linkers. Molecular docking and simulation studies revealed stable and high-affinity interactions. Furthermore, in-silico immune response graphs showed effective immune response generation. Finally, higher CAI value ensured high gene expression of vaccine in the host cell.
Conclusion: The designed MESV construct in the present in-silico study can be effective in generating an immune response against the yellow fever virus. Therefore, to prevent yellow fever, it can be an effective vaccine candidate. However, further downstream, in-vitro study is required.