Journal of Clinical and Biomedical Sciences
DOI: 10.58739/jcbs/v15i4.25.112
Year: 2025, Volume: 15, Issue: 4, Pages: 257-267
Original Article
Seyedeh Maryam Mousavi1*, Banafsheh Sadat Torabi2, Amin Hashemi3, Somayeh Mohammadi4, Masoud Faraji5, Mohsen Sadeghi6
1Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
2Department of Biology, Faculty of Sciences, Islamic Azad University, Damghan, Iran.
3Department of Microbiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran.
4Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Lahijan, Iran.
5Department of Microbiology, Faculty of Sciences, Islamic Azad University, Karaj, Iran.
6Department of Microbiology, Faculty of Advanced Sciences and Technology, Islamic Azad University, Tehran Medical Branch, Tehran, Iran.
*Corresponding Author
Email: [email protected]
Received Date:05 March 2025, Accepted Date:30 May 2025, Published Date:21 December 2025
Helicobacter pylori is a pathogen that has been consistently linked to gastrointestinal cancer, particularly gastric cancer. Reports indicate that this pathogen is resistant to antibiotic treatments, necessitating the development of new treatment strategies. Here, to design a vaccine against H. pylori immunoinformatic tools were used. For this purpose, five virulence proteins including Flagellin B (FlaB), Flagellin A (FlaA), Urease subunit beta (UreB), CagA and Vacuolating cytotoxin autotransporter (VacA) were selected. Antigenicity and allergenicity were evaluated, and the epitopes with the highest scores were chosen. Linkers were used to connect the epitopes and an extracellular domain of CTLA-4 was positioned on the N-terminal. Biochemical features were predicted with the ProtParam server, and the second structure was predicted with the Prabi server, while the third structure was predicted with the Robetta, Alphafold and I-TASSER servers. VaxiJen and AllerTOP servers were used to evaluate the vaccine's antigenicity and allergenicity, respectively. After confirming the structure of the designed vaccine, molecular docking was performed with the TLR5 molecule using AutoDock Vina software. The SnapGene tool was utilized to in silico cloning of the vaccine in pET-3a vector. To evaluate the efficacy of the recombinant vaccine, the multi-epitope gene of H. pylori was cloned into the pCDNA3.1 vector, and its expression was analyzed in the spleen tissue of BALB/c mice using RT-qPCR. It has been shown in the results that the vaccine designed can bind to the TLR5 molecule on the surface of immune cells. Despite being an antigen, this vaccine didn't have any allergenic properties. Immunoinformatic is a promising tool for designing various drugs and vaccines. The results demonstrated that the expression levels of TNF-α and IFN-γ were significantly increased in the vaccine-treated group compared to the control group (P < 0.05). The elevated expression of pro-inflammatory cytokines indicates that the designed DNA vaccine successfully induced a cellular immune response against H. pylori.
Keywords: H. pylori; Vaccine design; CTLA-4; Immunoinformatic; Gastric cancer
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Published By Sri Devaraj Urs Academy of Higher Education, Kolar, Karnataka
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