PENGEMBANGAN VAKSIN MULTI-EPITOP VIRUS DENGUE UNTUK POPULASI INDONESIA DENGAN PENDEKATAN REVERSE VACCINOLOGY

• Main Author: Gemilang, Kelvina
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/57139
• Institution: Institut Teknologi Bandung

Dengue virus (DENV) causes dengue hemorrhagic fever, that is the biggest health issue worldwide, including Indonesia. Variations in the four DENV serotypes pose challenges for vaccine development due to the presence of antibody dependent enhancement and original antigenic sin which increase the severity of the disease. The current commercial DENV vaccine is less effective because it can’t induce T cell response. Whereas the cross-react T cell response is protective between DENV serotypes. One method of making vaccines is Reverse Vaccinology (RV), which is the determination of potential vaccines using genomic and proteomic data from these organisms. This study aims to predict and select HTL, CTL, and BCL epitope DENV for the Indonesian population using a RV approach, construction and selection the multi-epitope vaccine DENV, predict and validate the three-dimensional structure model, predict interaction between vaccine constructs with TLRs and T cell epitopes with MHC receptors through docking analysis, and in silico cloning. The research method was carried out by collecting DENV protein data from NCBI as many as 62,249 sequences, merging each protein in all serotypes and selecting data so that it became 16,189 sequences, Multiple Sequence Alignment and diversity analysis using with conservation parameters ?98% and obtained 53 conserved epitope candidates. Then the prediction of CTL and HTL epitope with Human Leukocyte antigen (HLA) of the Indonesian people was carried out, as well as prediction population coverage and prediction of B cell epitope from HTL epitope. Epitope prediction results were selected and combined into one vaccine construct using 3 adjuvant variations and 3 epitope variations. After that, the 9 constructs were selected through characterization and comparison using multi-epitope vaccines that had been tested in vivo. The selected constructs were predicted for their secondary structure and tertiary structure, then validated using the Ramachandran plot, ERRAT score, and Z-score. Then, the docking analysis of vaccine constructs with TLR and epitope with MHC molecules was carried out. Finally, in silico cloning was carried out. Based on the results of the study, it was found that 5 CTL epitopes from NS5 protein and 9 HTL epitopes (with 3 overlapping epitopes with BCL epitopes) from protein NS5 and NS1 were antigenic, non-allergenic, non-toxic, hydrophilic, cytokine inducers, not similar to human and mouse proteins were then constructed into one vaccine construct. The results of the selection of vaccine constructs showed the 5 best constructs with a combination of 50S Ribosomal protein L7/L12 adjuvants Mycobacterium tuberculosis or CTB, linkers EAAAK and GPGPG, and containing 14 epitopes or 12 epitopes with 96,27% coverage in Indonesia’s population. These three constructs have a length of 312- 338 AA, were found antigenic, non-allergenic, non-toxic, hydrophilic, soluble, thermostable, have no transmembrane protein, and have good quality validated tertiary structures. All results of the docking analysis showed that the interaction between vaccine constructs and TLRs and T cell epitopes with MHC receptors could occur spontaneously (energetically feasible) and KV3 is the best construct based on docking analysis. In silico cloning of KV3 on the pRSET A plasmid was successful and has the potential to become a commercial DENV vaccine after further validation in vivo and in vitro.