Development of a Multiple Epitopes-Based Dengue Vaccine: An Immunoinformatics Approach and Insights From Pakistani Population Genetics.

Dengue fever poses a grave threat to public health worldwide, claiming numerous fatalities each year with tropical regions being particularly hard hit by dengue outbreaks. The multiple-epitope construct is tailored to the dengue virus's geographical prevalence and the genetics of the Pakistani population. 14 experimentally validated MHC-I and MHC-II epitope sequences, were employed to generate the variants by taking into account the conservancy in all serotypes. Subsequently, the binding affinities of each derived variant against the human leukocyte antigen alleles most common among the Pakistani population were analyzed. A total of three epitopes, two Class-I (GTSGSPIINR and RSWNTGFDW), and one Class-II (ILAPTRVVAAEMEEA), with a combined Pakistani population coverage of more than 73%, together with five linear B cell epitopes were used to create six possible multi-epitope fusion constructs. The molecular docking analysis indicated that two constructs demonstrated notable binding affinities for the most abundant HLA-A*11:01 in Pakistan. Additionally, molecular dynamics (MD) simulations identified one of the constructs as a promising therapeutic candidate. The vaccine construct selected from this analysis could aid in future vaccine design for the dengue virus following further in vitro test validation and in vivo studies to investigate its immune protection capacity.