Immunogenic Potency of a Chimeric Protein Comprising InvH and IpaD against Salmonella and Shigella spp
Abstract
Shigella and Salmonella cause serious problems in many subjects, including young children and the elderly, especially in developing countries. Chimeric proteins carrying immunogens increase immune response. In-silico tools are applied to design vaccine candidates. Invasion plasmid antigens D (ipaD) gene is one of the Shigella virulence factors. The N-terminal region of the IpaD plays a significant role in invading the host cell. Invasion protein H (invH) gene plays important role in bacterial adherence and entry into epithelial cells. A recombinant chimeric construct, containing IpaD and InvH was designed and used as a vaccine candidate against Shigella and Salmonella enteritidis.
After bioinformatics assessments, the construct was designed, synthesized, and expressed in E.coli. Chimeric protein, IpaD, and InvH were purified with Ni-NTA chromatography. Purified proteins were confirmed with western blotting and then were injected into separate mice groups. The antibody titer was estimated with an enzyme-linked immunosorbent assay (ELISA). Mice were challenged with 10, 100, and 1000 LD50 of Salmonella, and the sereny test was performed for Shigella.
The Codon adaptation index of the chimeric gene was increased to 0.84. Validation results showed that 97.9% of residues lie in the favored or additional allowed region of the Ramachandran plot. A significant antibody rise was observed in all test groups. The immunized mice with chimer and InvH could tolerate 100 LD50 of Salmonella. In the sereny test, the application of bacteria treated with immunized mice sera of both antigens showed no infection in Guinea pigs' eyes.
The recombinant protein could protect animal models against Salmonella and Shigella and therefore can be considered as a suitable vaccine candidate against these two pathogens.
2. Ranjbar R, Naghoni A, Afshar D, Nikkhahi F, Mohammadi M. Rapid Molecular Approach for Simultaneous Detection of Salmonella spp., Shigella spp., and Vibrio cholera. Osong Public Health Res Perspect. 2016;7(6):373-7.
3. Shakoor S, Zaidi AK, Hasan R. Tropical bacterial gastrointestinal infections. Infect Dis Clin North Am. 2012;26(2):437-53.
4. Tennant SM, MacLennan CA, Simon R, Martin LB, Khan MI. Nontyphoidal salmonella disease: Current status of vaccine research and development. Vaccine. 2016;34(26):2907-10.
5. Dekker JP, Frank KM. Salmonella, Shigella, and yersinia. Clin Lab Med. 2015;35(2):225-46.
6. Saadati M, Heiat M, Nazarian S, Barati B, Honari H, Doroudian M, et al. Cloning and Expression of N-terminal Region of IpaD from Shigella dysenteriae in E. coli. J Paramed Sci. 2010;1(4):12-7.
7. Sukhan A, Kubori T, Wilson J, Galán JE. Genetic analysis of assembly of theSalmonella enterica serovar Typhimurium type III secretion-associated needle complex. J Bacteriol. 2001;183(4):1159-67.
8. Burkinshaw BJ, Strynadka NC. Assembly and structure of the T3SS. Biochim Biophys Acta Mol Cell Res. 2014;1843(8):1649-63.
9. Altmeyer RM, McNern JK, Bossio J, Rosenshine I, Finlay BB, Galán JE. Cloning and molecular characterization of a gene involved in Salmonella adherence and invasion of cultured epithelial cells. Mol Microbiol. 1993;7(1):89-98.
10. Salamaszynska-Guz A, Klimuszko D. Strategies of constructing recombinant vaccines. Pol J Vet Sci. 2009;12(1):133-9.
11. Alerasol M, Gargari SLM, Nazarian S, Bagheri S. Immunogenicity of a fusion protein comprising coli surface antigen 3 and labile B subunit of enterotoxigenic Escherichia coli. Iranian biomedical journal. 2014;18(4):212.
12. Dehghania B, Hashempoura T, Hasanshahia Z, Rasoolib I. Immunogenicity, antigenicity and epitope mapping of Salmonella InvH protein: An in silico study. J CURR BIOMED REP. 2020;1(1).
13. Jneid B, Rouaix A, Féraudet-Tarisse C, Simon S. SipD and IpaD induce a cross-protection against Shigella and Salmonella infections. PLoS Negl Trop Dis. 2020;14(5):e0008326.
14. Nosrati M, Hajizade A, Nazarian S, Amani J, Vansofla AN, Tarverdizadeh Y. Designing a multi-epitope vaccine for cross-protection against Shigella spp: An immunoinformatics and structural vaccinology study. Mol Immunol. 2019;116:106-16.
15. Chen X, Zaro JL, Shen W-C. Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev. 2013;65(10):1357-69.
16. Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res. 2007;35(suppl_2):W407-W10.
17. Lovell SC, Davis IW, Arendall III WB, De Bakker PI, Word JM, Prisant MG, et al. Structure validation by Cα geometry: ϕ, ψ and Cβ deviation. Proteins. 2003;50(3):437-50.
18. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. nature. 1970;227(5259):680.
19. Dehghani B, Rasooli I, Gargari SLM, Nadooshan MRJ, Owlia P, Nazarian S. Immunogenicity of Salmonella enterica serovar Enteritidis virulence protein, InvH, and cross-reactivity of its antisera with Salmonella strains. Microbiol Res. 2013;168(2):84-90.
20. Bollag DM, Rozycki MD, Edelstein SJ. Protein methods: Wiley-Liss New York; 1996.
21. Council NR. Guide for the care and use of laboratory animals: National Academies Press; 2010.
22. Hartman AB, Powell CJ, Schultz CL, Oaks EV, Eckels KH. Small-animal model to measure efficacy and immunogenicity of Shigella vaccine strains. Infect Immun. 1991;59(11):4075-83.
23. Hanson CE, Ruble GR, Essiet I, Hartman AB. Effects of Buprenorphine on Immunogenicity and Protective Efficacy in the Guinea Pig Keratoconjunctivitis Model (Sereny Test). Comparative Medicine. 2001;51(3):224-9.
24. Cai X, Zhang J, Chen M, Wu Y, Wang X, Chen J, et al. The effect of the potential PhoQ histidine kinase inhibitors on Shigella flexneri virulence. PLoS One. 2011;6(8):e23100.
25. Ren J, Wen L, Gao X, Jin C, Xue Y, Yao X. DOG 1.0: illustrator of protein domain structures. Cell Res. 2009;19(2):271.
26. Robinson J, Burke V, Worthy P, Beaman J, Wagener L. Media for isolation of Aeromonas spp. from faeces. J Med Microbiol. 1984;18(3):405-11.
27. Kubler-Kielb J, Vinogradov E, Mocca C, Pozsgay V, Coxon B, Robbins JB, et al. Immunochemical studies of Shigella flexneri 2a and 6, and Shigella dysenteriae type 1 O-specific polysaccharide-core fragments and their protein conjugates as vaccine candidates. Carbohydr Res. 2010;345(11):1600-8.
28. Ochoa J, Irache JM, Tamayo I, Walz A, DelVecchio VG, Gamazo C. Protective immunity of biodegradable nanoparticle-based vaccine against an experimental challenge with Salmonella Enteritidis in mice. Vaccine. 2007;25(22):4410-9.
29. Hansson M, Nygren P-Ak, Sta˚hl S. Design and production of recombinant subunit vaccines. Biotechnology and Applied Biochemistry. 2000;32(2):95-107.
30. Hayat S-MG, Gargari S-LM, Nazarian S. Construction and immunogenic properties of a chimeric protein comprising CfaE, CfaB and LTB against Enterotoxigenic Escherichia coli. Biologicals. 2016;44(6):503-10.
31. Martinez-Becerra FJ, Scobey M, Harrison K, Choudhari SP, Quick AM, Joshi SB, et al. Parenteral immunization with IpaB/IpaD protects mice against lethal pulmonary infection by Shigella. Vaccine. 2013;31(24):2667-72.
32. Nazarian S, Gargari SLM, Rasooli I, Amani J, Bagheri S, Alerasool M. An in silico chimeric multi subunit vaccine targeting virulence factors of enterotoxigenic Escherichia coli (ETEC) with its bacterial inbuilt adjuvant. J Microbiol Methods. 2012;90(1):36-45.
33. Arai R, Ueda H, Kitayama A, Kamiya N, Nagamune T. Design of the linkers which effectively separate domains of a bifunctional fusion protein. Protein Eng. 2001;14(8):529-32.
34. Nazarian S, Gargari SLM, Rasooli I, Hasannia S, Pirooznia N. A PLGA-encapsulated chimeric protein protects against adherence and toxicity of enterotoxigenic Escherichia coli. Microbiol Res. 2014;169(2-3):205-12.
35. Amani J, Salmanian AH, Rafati S, Mousavi SL. Immunogenic properties of chimeric protein from espA, eae and tir genes of Escherichia coli O157: H7. Vaccine. 2010;28(42):6923-9.
Files | ||
Issue | Vol 20 No 4 (2021) | |
Section | Original Article(s) | |
DOI | https://doi.org/10.18502/ijaai.v20i4.6958 | |
Keywords | ||
InvH protein IpaD protein Recombinant proteins Salmonella Shigella |
Rights and permissions | |
![]() |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |