Volume 27, Issue 6 (Iran South Med J 2025)
Iran South Med J 2025, 27(6): 418-432 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Samiei-Abianeh H, Tat M, Bolandian M, Arianzad S A, Salimian J. In Silico Analysis of Immunogenic Epitopes in the VP2 Protein of Canine Parvovirus and Codon Optimization for Multivalent Vaccine Design. Iran South Med J 2025; 27 (6) :418-432
URL: http://ismj.bpums.ac.ir/article-1-2169-en.html
URL: http://ismj.bpums.ac.ir/article-1-2169-en.html
Hossein Samiei-Abianeh1 
, Mahdi Tat2 , Masoumeh Bolandian2 , Seyed Akbar Arianzad3 , Jafar Salimian *4
, Mahdi Tat2 , Masoumeh Bolandian2 , Seyed Akbar Arianzad3 , Jafar Salimian *4
1- Department of Medical Biotechnology and Nanotechnology, School of Medicine, Mashhad University of Medical Sciences Mashhad, Iran
Department of Biology, School of Basic Sciences, Imam Hossein University, Tehran, Iran
2- Applied Virology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
3- Department of Microbiology, School of Scince, Islamic Azad Universty, Karaj Branch, Karaj, Iran
4- Applied Virology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran ,jafar.salimian@gmail.com
Department of Biology, School of Basic Sciences, Imam Hossein University, Tehran, Iran
2- Applied Virology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
3- Department of Microbiology, School of Scince, Islamic Azad Universty, Karaj Branch, Karaj, Iran
4- Applied Virology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran ,
Abstract: (188 Views)
Background: Canine parvovirus (CPV) is one of the major agents of severe viral gastroenteritis in dogs. Although vaccines are available, the emergence of new serotypes and mutations in the VP2 protein can compromise vaccine efficacy. Therefore, identifying both conserved and serotype-specific immunogenic regions is crucial for the design of recombinant vaccines. This study aimed to design and optimize a recombinant vaccine based on conserved immunogenic regions of the VP2 protein from different CPV serotypes.
Materials and Methods: Amino acid sequences of the VP2 protein from four CPV serotypes were retrieved from the NCBI database and aligned using Clustal Omega. Structural and immunological features were analyzed using tools such as SignalP, TOPOCONS, and DIpro. Protein solubility, secondary and tertiary structures, and physicochemical properties were evaluated using Protein-Sol, I-TASSER, and ProtParam. The coding sequences were optimized for expression in E. coli and the mRNA secondary structure was analyzed using MFOLD.
Results: Multiple sequence alignment identified five key amino acid differences in the 285–450 region of VP2. Tertiary structure analysis showed that this region forms two surface-exposed loops, with the mutated residues at positions 297 and 426 located at the tips. Deletion of the first 20 amino acids improved structural stability, lowering the instability index to 29. Codon optimization increased CAI value to 0.79 and decreased mRNA free energy to –597 kcal/mol.
Conclusion: A chimeric recombinant vaccine targeting the 285–450 region of the VP2 protein across CPV serotypes shows promise as a candidate for broad-spectrum protection. Despite in silico results, experimental validation of protein expression and immune response in animal models is essential.
Materials and Methods: Amino acid sequences of the VP2 protein from four CPV serotypes were retrieved from the NCBI database and aligned using Clustal Omega. Structural and immunological features were analyzed using tools such as SignalP, TOPOCONS, and DIpro. Protein solubility, secondary and tertiary structures, and physicochemical properties were evaluated using Protein-Sol, I-TASSER, and ProtParam. The coding sequences were optimized for expression in E. coli and the mRNA secondary structure was analyzed using MFOLD.
Results: Multiple sequence alignment identified five key amino acid differences in the 285–450 region of VP2. Tertiary structure analysis showed that this region forms two surface-exposed loops, with the mutated residues at positions 297 and 426 located at the tips. Deletion of the first 20 amino acids improved structural stability, lowering the instability index to 29. Codon optimization increased CAI value to 0.79 and decreased mRNA free energy to –597 kcal/mol.
Conclusion: A chimeric recombinant vaccine targeting the 285–450 region of the VP2 protein across CPV serotypes shows promise as a candidate for broad-spectrum protection. Despite in silico results, experimental validation of protein expression and immune response in animal models is essential.
Type of Study: Original |
Subject:
Virology
Received: 2025/04/18 | Accepted: 2025/06/30 | Published: 2025/07/22
Received: 2025/04/18 | Accepted: 2025/06/30 | Published: 2025/07/22
References
1. Parrish CR. Pathogenesis of feline panleukopenia virus and canine parvovirus. Baillieres Clin Haematol 1995; 8(1): 57-71. [Article]
2. Houston DM, Ribble CS, Head LL. Risk factors associated with parvovirus enteritis in dogs : 283 cases (1982-1991). J Am Vet Med Assoc 1996; 208(4): 542-46. [Article]
3. Truyen U. Evolution of canine parvovirus--a need for new vaccines?. Vet Microbiol 2006; 117(1): 9-13. [Article]
4. Gallo Calderón M, Wilda M, Boado L, et al. Study of canine parvovirus evolution: comparative analysis of full-length VP2 gene sequences from Argentina and international field strains. Virus Genes 2012; 44(1): 32-9. [DOI]
5. Parrish CR, Have P, Foreyt WJ, et al. The global spread and replacement of canine parvovirus strains. J Gen Virol 1988; 69(Pt 5): 1111-16. [DOI]
6. Buonavoglia C, Martella V, Pratelli A, et al. Evidence for evolution of canine parvovirus type 2 in Italy. J Gen Virol 2001; 82(Pt 12): 3021-25. [DOI]
7. Miranda C, Thompson G. Canine parvovirus: the worldwide occurrence of antigenic variants. J Gen Virol 2016; 97(9): 2043-57. [DOI]
8. Decaro N, Buonavoglia C, Barrs VR. Canine parvovirus vaccination and immunisation failures: Are we far from disease eradication?. Vet Microbiol 2020; 247: 108760. [Article]
9. Hoang M, Lin WH, Le VP, et al. Molecular epidemiology of canine parvovirus type 2 in Vietnam from November 2016 to February 2018. Virol J 2019; 16(1): 52. [DOI]
10. Chiang SY, Wu HY, Chiou MT, et al. Identification of a novel canine parvovirus type 2c in Taiwan. Virol J 2016; 13(1): 160. [DOI]
11. Woolford L, Crocker P, Bobrowski H, et al. Detection of the Canine Parvovirus 2c Subtype in Australian Dogs. Viral Immunol 2017; 30(5): 371-6. [DOI]
12. Hernández-Blanco B, Catala-López F. Are licensed canine parvovirus (CPV2 and CPV2b) vaccines able to elicit protection against CPV2c subtype in puppies?: A systematic review of controlled clinical trials. Vet Microbiol 2015; 180(1-2): 1-9. [Article]
13. Larson LJ, Schultz RD. Do two current canine parvovirus type 2 and 2b vaccines provide protection against the new type 2c variant?. Vet Ther 2008; 9(2): 94-101. [Article]
14. Spibey N, Greenwood NM, Sutton D, et al. Canine parvovirus type 2 vaccine protects against virulent challenge with type 2c virus. Vet Microbiol 2008; 128(1-2): 48-55. [Article]
15. Dahiya SS, Saini M, Kumar P, et al. Immunogenicity of a DNA-launched replicon-based canine parvovirus DNA vaccine expressing VP2 antigen in dogs. Res Vet Sci 2012; 93(2): 1089-97. [Article]
16. López de Turiso JA, Cortés E, Martínez C, et al. Recombinant vaccine for canine parvovirus in dogs. J Virol 1992; 66(5): 2748-53. [DOI]
17. Inthong N, Kaewmongkol S, Meekhanon N, et al. Expression of recombinant 35 kDa fragment of VP2 protein of canine parvovirus using Escherichia coli expression system. Vet World 2021; 14(6): 1682-1688. [Article]
18. Wikoff WR, Wang G, Parrish CR, et al. The structure of a neutralized virus: canine parvovirus complexed with neutralizing antibody fragment. Structure 1994; 2(7): 595-607. [Article]
Send email to the article author
| Rights and Permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
