In Silico Characterization and Epitope Mapping of Echinococcus granulosus  Annexin Protein: Novel Insights for Vaccine Design

Background: A neglected zoonosis, cystic echinococcosis (CE) is the most common diseas in the developing nations worldwide. Vaccination is helpful in preventing the disease. Predicting main biochemical properties of the E. granulosus annexin (ANX) and its possible B-cell and human leukocyte antigen (HLA)-binding epitopes as a viable vaccine candidate was the goal of the current study.
Methods: This study was done in Neyshabur University of Medical Sciences, Neyshabur, Iran. Predictions about transmembrane domain, subcellular localization, post-translational modification (PTM) sites, physico-chemical, antigenic, and allergenic profiles, secondary and 3D structure, tertiary model refinement, and validations were done using online servers.
Results: The cytoplasmic 79.05 kDa protein was non-allergenic, hydrophilic (GRAVY: -0.490), stable (instability: 39.30), with improved thermotolerance (aliphatic: 80.07) and 122 post-translational modification sites. The secondary structure mostly included helices and extended strands. The 3D model was generated using Robetta server (confidence: 0.59) and was refined and validated subsequently. Shared B-cell epitopes were discovered using ElliPro, ABCpred and SVMTriP servers with antigenicity, allergenicity, and solubility screening. Moreover, multiple human and mouse MHC-binding epitopes were predicted and screened in E. granulosus ANX.
Conclusion: This work offers a foundation for further investigation regarding designing an effective vaccination against CE. Further empirical research using examined protein alone or in conjunction with other antigens/epitopes is needed in the future.

1. Wen H, Vuitton L, Tuxun T, et al. Echinococcosis: advances in the 21st century. Clin Microbiol Rev. 2019;32(2):e00075-18.
2. Khalkhali H, Foroutan M, Khademvatan S, et al. Prevalence of cystic echinococcosis in Iran: a systematic review and meta-analysis. J Helminthol. 2018;92(3):260-8.
3. McManus DP, Gray DJ, Zhang W, Yang YJB. Diagnosis, treatment, and management of echinococcosis. BMJ. 2012;344:e3866.
4. Budke CM, Carabin H, Ndimubanzi PC, et al. A systematic review of the literature on cystic echinococcosis frequency worldwide and its associated clinical manifestations. Am J Trop Med Hyg. 2013;88(6):1011-27.
5. Budke CM, Deplazes P, Torgerson PR. Global socioeconomic impact of cystic echinococcosis. Emerg Infect Dis. 2006;12(2):296-303.
6. Larrieu E, Gavidia CM, Lightowlers MW. Control of cystic echinococcosis: Background and prospects. Zoonoses Public Health. 2019;66(8):889-99.
7. Pourseif MM, Moghaddam G, Saeedi N, et al. Current status and future prospective of vaccine development against Echinococcus granulosus. Biologicals. 2018;51:1-11.
8. Lightowlers M, Colebrook A, Gauci C, Gauci S, Kyngdon C, Monkhouse J, et al. Vaccination against cestode parasites: anti-helminth vaccines that work and why. Vet Parasitol. 2003;115(2):83-123.
9. Vercruysse J, Knox DP, Schetters TP, Willadsen P. Veterinary parasitic vaccines: pitfalls and future directions. Trends Parasitol. 2004;20(10):488-92.
10. Craig P, Hegglin D, Lightowlers M, Torgerson PR, Wang Q. Echinococcosis: control and prevention. Adv Parasitol. 2017; 96: 55-158.
11. Siles-Lucas M, Sánchez-Ovejero C, González-Sánchez M, et al. Isolation and characterization of exosomes derived from fertile sheep hydatid cysts. Vet Parasitol. 2017;236:22-33.
12. Virginio VG, Monteiro KM, Drumond F, et al. Excretory/secretory products from in vitro-cultured Echinococcus granulosus protoscoleces. Mol Biochem Parasitol. 2012;183(1):15-22.
13. Zeghir-Bouteldja R, Polomé A, Bousbata S, Touil-Boukoffa C. Comparative proteome profiling of hydatid fluid from Algerian patients reveals cyst location-related variation in Echinococcus granulosus. Act Trop. 2017;171:199-206.
14. Song H, He X, Du X, et al. Molecular characterization and expression analysis of annexin B3 and B38 as secretory proteins in Echinococcus granulosus. Parasit Vectors. 2021;14:103.
15. Zhu Y, He Y, Yin Z, et al. Enhanced Immune Response Against Echinococcus granulosus Through a CTLA-4/B7 Affinity-Based Vaccine. Vaccines (Basel). 2024;12(12):1440.
16. Gasteiger E, Hoogland C, Gattiker A, et al. Protein identification and analysis tools on the ExPASy server. The proteomics protocols handbook. 1st Ed. St. Loius: Springer; 2005. p. 571-607.
17. Heo L, Park H, Seok C. GalaxyRefine: protein structure refinement driven by side-chain repacking. Nucleic Acids Res. 2013; 41(Web Server issue):W384-8.
18. Majid M, Andleeb S. Designing a multi-epitopic vaccine against the enterotoxigenic Bacteroides fragilis based on immunoinformatics approach. Sci Rep. 2019;9(1): 19780.
19. Ponomarenko J, Bui H-H, Li W, et al. ElliPro: a new structure-based tool for the prediction of antibody epitopes. BMC Bioinformatics. 2008;9(1):514.
20. Greenbaum J, Sidney J, Chung J, et al. Functional classification of class II human leukocyte antigen (HLA) molecules reveals seven different supertypes and a surprising degree of repertoire sharing across supertypes. Immunogenetics. 2011; 63(6):325-35.
21. Song X, Hu D, Zhong X, et al. Characterization of a Secretory Annexin in Echinococcus granulosus. Am J Trop Med Hyg. 2016;94(3):626-33.
22. Majidiani H, Soltani S, Ghaffari AD, et al. In-depth computational analysis of calcium-dependent protein kinase 3 of Toxoplasma gondii provides promising targets for vaccination. Clin Exp Vaccine Res. 2020;9(2):146-158.
23. Seadawy MG, Zekri ARN, Saeed AA, et al. Candidate Multi-Epitope Vaccine against Corona B.1.617 Lineage: In Silico Approach. Life (Basel). 2022;12(11):1715.
24. Walsh C. Posttranslational modification of proteins: expanding nature's inventory. 1st Ed. Greenwood Village (CO): Roberts and Company Publishers; 2006. 576 p.
25. Lee T-Y, Hsu JB-K, Chang W-C, Wang T-Y, Hsu P-C, Huang H-D. A comprehensive resource for integrating and displaying protein post-translational modifications. BMC Res Notes. 2009;2:111.
26. Hansson M, Nygren PAk, Sta˚ hl SJB, biochemistry a. Design and production of recombinant subunit vaccines. Biotechnol Appl Biochem. 2000;32(2):95-107.
27. Yadav G, Rao R, Raj U, Varadwaj PK. Computational modeling and analysis of prominent T-cell epitopes for assisting in designing vaccine of ZIKA virus. J Appl Pharm Sci. 2017;7(08):116-22.
28. Shaddel M, Ebrahimi M, Tabandeh MR. Bioinformatics analysis of single and multi-hybrid epitopes of GRA-1, GRA-4, GRA-6 and GRA-7 proteins to improve DNA vaccine design against Toxoplasma gondii. J Parasit Dis. 2018;42(2):269-76.
29. Zhang W, Wen H, Li J, Lin R, McManus DPJC. Immunology and immunodiagnosis of cystic echinococcosis: an update. Clin Dev Immunol. 2012;2012: 101895.

Files	XML PDF (2MB)
Issue	Vol 20 No 3 (2025)
Section	Original Article(s)
Keywords
Immunoinformatics Echinococcus granulosus Annexin Immunization

Rights and permissions
	This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

How to Cite

Khodadadi A, Rastgoo MH, Ghiyasipour E, Moradmandpaeinroudposhti R, Zandieh MA, Zahirinia F, Haghdadijoo M, Esfandani A, Rajabi R, Marashi A, Dadboud MA, Majidiani H, Rahmanian P. In Silico Characterization and Epitope Mapping of Echinococcus granulosus Annexin Protein: Novel Insights for Vaccine Design. Iran J Parasitol. 2025;20(3):378-388.

Vancouver

Download Citation