Designing a Multi-Epitope Vaccine Against HPV 16, 18, 33, and 45 Targeting L1 and E7 Proteins: An Immunoinformatics Approach for Cervical Cancer Prevention and Therapy

Cervical cancer, induced by human papillomavirus (HPV), ranks as the fourth most prevalent malignancy among women globally. Unfortunately, existing prophylactic vaccines lack therapeutic efficacy. This study aimed to design a multi-epitope vaccine targeting the L1 and E7 proteins of HPV 16, 18, 33, and 45, with both preventive and therapeutic potential. Epitopes predicted using Immune Epitope Database (IEDB) and ABCpred were screened via immunoinformatics tools for antigenicity, immunogenicity, safety, conservancy, population coverage, and homology, and appropriate epitopes were assembled into a vaccine with suitable linkers and a 50-S L7/L12 adjuvant. The modeled and optimized vaccine was immunogenic, antigenic, safe, and displayed favorable physicochemical and solubility properties. Docking studies using ClusPro 2.0 and HDOCK indicated robust interactions between the vaccine and toll-like receptors TLR2/TLR4, and molecular dynamics simulations with Desmond validated the structural stability. Furthermore, molecular mechanics with generalized born and surface area solvation (MM/GBSA) analysis employing HawkDock showed favorable binding free energies of -82.86 and -76.72 kcal/mol, respectively. The vaccine's potential efficacy was demonstrated by C-IMMSIM immune simulations, which revealed robust and long-lasting cellular and humoral responses, and also strong cytokine production. Finally, codon optimization for <i>Escherichia coli</i> K12 using JCat yielded a guanine-cytosine content of 50.69% and a Codon Adaptation Index of 0.97, and <i>in silico</i> cloning into pET28a(+) using SnapGene confirmed high expression potential. Our results indicate that the designed vaccine is a viable candidate for both preventive and therapeutic measures against high-risk HPV, requiring additional laboratory and animal studies.