Background: Influenza D Virus (IDV) presents a possible threat to animal and human health, necessitating the development of effective vaccines. Although no human illness linked to IDV has been reported, the possibility of human susceptibility to infection remains uncertain. Hence, there is a need for an animal vaccine to be designed. Such a vaccine will contribute to preventing and controlling IDV outbreaks and developing effective countermeasures against this emerging pathogen. This study, therefore, aimed to design an mRNA vaccine construct against IDV using immunoinformatic methods and evaluate its potential efficacy. Methods: A comprehensive methodology involving epitope prediction, vaccine construction, and structural analysis was employed. Viral sequences from six continents were collected and analyzed. A total of 88 Hemagglutinin Esterase Fusion (HEF) sequences from IDV isolates were obtained, of which 76 were identified as antigenic. Different bioinformatics tools were used to identify preferred CTL, HTL, and B-cell epitopes. The epitopes underwent thorough analysis, and those that can induce a lasting immunological response were selected for the construction. Results: The vaccine prototype comprised nine epitopes, an adjuvant, MHC I-targeting domain (MITD), Kozaq, 3′ UTR, 5′ UTR, and specific linkers. The mRNA vaccine construct exhibited antigenicity, non-toxicity, and non-allergenicity, with favourable physicochemical properties. The secondary and tertiary structure analyses revealed a stable and accurate vaccine construct. Molecular docking simulations also demonstrated strong binding affinity with toll-like receptors. Conclusions: The study provides a promising framework for developing an effective mRNA vaccine against IDV, highlighting its potential for mitigating the global impact of this viral infection. Further experimental studies are needed to confirm the vaccine’s efficacy and safety.

BACKGROUND: Nasopharyngeal Carcinoma(NPC) is a rare tumor in the world, but high in some countries such as southern China, Mediterranean Africa and Middle East. According to studies, Epstein-Barr virus is affected more aggressive in NPC. In NPC, EBV-miR-BARTs have been demonstrated, especially EBV-miR-BART17 have relation to β-catenin activity but the studies are still unclear. Peroxiredoxin 1 (Prdx1) is an antioxidant enzyme and is highly susceptible to oxidative stress. METHODS: To study the roles of Prdx1 and EBV-miR-BART17-5p in NPC C666-1 and HK1 cells, it is transfected to siRNA of Prdx1 and then it is measured using RT-PCR for RNA level and western blot for protein level. Colony forming assay and Wound-healing assay were used to detect the cell proliferation in NPC cells. RESULT: EBV-miR-BART17-5p expression was higher in C666-1 than HK1 cells. The expression of Prdx1 is different between C666-1 and HK1 cells because of EBV genes. EBV-miR-BART17 expression was found to relate to cell proliferation through targeting NF-κB signaling pathway. CONCLUSION: These data suggested that Prdx1 and EBV-miR-BART17-5p play roles in inducing β-catenin activity in NPC cells. Our findings suggest that the regulating of Prdx1 is an important therapeutic target for NPC.