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Dengue fever (DF) is a major public health concern that affects approximately 3.9 billion people each year globally. However, there is no vaccine or drug available to deal with DVI. Dengue virus consists of four distinct serotypes (DENV1-4), each raising a different immunological response. In the present study, we designed a tetravalent subunit multi-epitope vaccine, targeting proteins including the structural protein envelope domain III (EDIII), precursor membrane proteins (prM), and a non-structural protein (NS1) from each serotype by employing an immunoinformatic approach. Only conserved sequences obtained through a multiple sequence alignment were used for epitope mapping to ensure efficacy against all serotypes. The epitopes were shortlisted based on an IC50 value &amp;lt;50, antigenicity, allergenicity, and a toxicity analysis. In the final vaccine construct, overall, 11 B-cell epitopes, 10 HTL epitopes, and 10 CTL epitopes from EDIII, prM, and NS1 proteins targeting all serotypes were selected and joined via KK, AAY, and GGGS linkers, respectively. We incorporated a 45-amino-acid-long B-defensins adjuvant in the final vaccine construct for a better immunogenic response. The vaccine construct has an antigenic score of 0.79 via VaxiJen and is non-toxic and non-allergenic. Our refined vaccine structure has a Ramachandran score of 96.4%. The vaccine has shown stable interaction with TLR3, which has been validated by 50 ns of molecular dynamics (MD) simulation. Our findings propose that a designed multi-epitope vaccine has substantial potential to elicit a strong immune response against all dengue serotypes without causing any adverse effects. Furthermore, the proposed vaccine can be experimentally validated as a probable vaccine, suggesting it may serve as an effective preventative measure against dengue virus infection.<\/jats:p>","DOI":"10.3389\/fimmu.2023.1273838","type":"journal-article","created":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T10:32:08Z","timestamp":1700217128000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":12,"title":["Development of a tetravalent subunit vaccine against dengue virus through a vaccinomics approach"],"prefix":"10.3389","volume":"14","author":[{"given":"Amina","family":"Basheer","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Syed Babar","family":"Jamal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Badr","family":"Alzahrani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Faheem","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,11,17]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1093\/jtm\/taab146","article-title":"Global burden for dengue and the evolving pattern in the past 30 years","volume":"28","author":"Yang","year":"2021","journal-title":"J Travel Med"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3855\/jidc.1017","article-title":"Dengue fever in the Indian subcontinent: An overview","volume":"5","author":"Raheel","year":"2011","journal-title":"J Infect Dev Ctries"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-010-0488-1","article-title":"A molecular evaluation of dengue virus pathogenesis and its latest vaccine strategies","volume":"38","author":"Faheem","year":"2011","journal-title":"Mol Biol Rep"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0002056","article-title":"Use of expansion factors to estimate the burden of dengue in southeast asia: A systematic analysis","volume":"7","author":"Undurraga","year":"2013","journal-title":"PloS Negl Trop Dis"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003306","article-title":"Approaches to refining estimates of global burden and economics of dengue","volume":"8","author":"Shepard","year":"2014","journal-title":"PloS Negl Trop Dis"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2016.04.008","article-title":"Progress and prospects on DENV protease inhibitors","volume":"117","author":"Timiri","year":"2016","journal-title":"Eur J Med Chem"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.01651","article-title":"The dual role of the antibody response against the flavivirus non-structural protein 1 (NS1) in protection and immuno-pathogenesis","volume":"10","author":"Reyes-Sandoval","year":"2019","journal-title":"Front Immunol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1700029","article-title":"Antibody epitopes identified in critical regions of dengue virus nonstructural 1 protein in mouse vaccination and natural human infections","volume":"198","author":"Hertz","year":"2017","journal-title":"J Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1317350111","article-title":"Dengue virus envelope protein domain I\/II hinge determines long-lived serotype-specific dengue immunity","volume":"111","author":"Messer","year":"2014","journal-title":"Proc Natl Acad Sci USA"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1038\/cmi.2015.76","article-title":"Guiding dengue vaccine development using knowledge gained from the success of the yellow fever vaccine","volume":"13","author":"Liang","year":"2016","journal-title":"Cell. 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