Abstract
The findings underscore the critical role of Epstein-Barr virus (EBV) in the onset of various cancers. In response to the lack of effective treatments or vaccines for EBV infection, this investigation employed immunoinformatics approaches to develop a potent vaccine targeting multiple epitopes of the EBV glycoprotein 350 (Gp350), a key surface protein. Utilizing computational methods, we designed a comprehensive multi-epitope vaccine featuring 11 CTL and HTL epitopes, totaling 324 amino acids and covering five distinct EBV strains such as B95-8, P3HR-1, GD1, AG876, and Akata. To enhance immunogenicity, the 50S ribosomal protein L7/L12 (rplL) was included as an adjuvant at the vaccine's N-terminal. The vaccine was evaluated for its physicochemical and immunological properties, demonstrating stability, potency, solubility, hydrophilicity, non-allergenicity, and non-toxicity. Molecular docking studies have shown that the vaccine interacts with Toll-like receptor 4 (TLR4). Simulations performed using GROMACS confirmed the stability of the system over 100ns. Immune simulations indicated that the vaccine elicited robust humoral and cellular responses, activating both innate and adaptive immunity. The findings indicate that the multi-epitope vaccine is highly immunogenic and shows significant potential for further experimental validation.