Antigenic and immunogenic evaluation of Helicobacter pylori FlaA epitopes

Document Type : Original Article

Authors

1 Department of Medical Biotechnology, School of Medicine, Arak University of Medical Sciences, Arak, Iran

2 Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran

3 Molecular and Medicine Research Center, Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran

Abstract

Objective(s): Helicobacter pyloriare among most common human pathogens affecting at least half of the world’s population. Mobility is one of the important primary factors in bacterial colonization and invasion. The purpose of this research is cloning, expression, and purification of FlaA protein specific epitopes in order to evaluate their antigenicity and immunogenicity.
Materials and Methods: The antigenic region of the flaA gene was bioinformatically predicted using Epitope mapping software’s and the predicted epitopes were expressed in a prokaryotic expression vector. The antigen was injected into the animal model (mice BALB/c) and some indicators including IgG1, IgG2a, IgA, IFN-γ, and IL 5 were measured.
Results: The immunogenicity studies in animal models by measuring serum antibodies (IgG1, IgG2a, and IgA) and cytokines (IFN-γ and IL5) revealed that the rFlaA induces a proper immune response in animal models.
Conclusion: The recombinant FlaA protein is antigenic and immunogenic. Therefore, it might be used in order to design of specific diagnostic kits and recombinant vaccines against H. pylori.

Keywords


1. Kalali B, Mejías-Luque R, Javaheri A, Gerhard M. Helicobacter pylori virulence factors: influence on immune system and pathology. Mediators Inflamm 2014; 2014:426309.
2. Ruggiero P, Censini S. Helicobacter pylori: A Brief History of a still lacking vaccine. Diseases 2014; 2:187-208.
3 .Petersen AM, Krogfelt KA. Helicobacter pylori: an invading microorganism? A review. FEMS Immunol Med Microbiol 2003; 36:117-126.
4. Dubois A, Borén T. Helicobacter pylori is invasive and it may be a facultative intracellular organism. Cell Microbiol 2007; 9:1108-1116.
5. Chu YT, Wang YH, Wu JJ, Lei HY. Invasion and multiplication of Helicobacter pylori in gastric epithelial cells and implications for antibiotic resistance. Infect Immun 2010; 78:10:4157-4165.
6. Wang YH, Wu JJ, Lei HY. The autophagic induction in Helicobacter pylori infected macrophage. Exp Biol Med 2009; 234:171-180.
7. Mehmood K, Hasan F. Construction and use of a prokaryotic expression system for Helicobacter pylori AhpC. BMC Res Notes 2012; 5:328.
8. Ghasemi-Kebria F, Ghaemi E, Azadfar S, Roshandel G. Epidemiology of Helicobacter pylori infection among Iranian children. Arab J Gastroenterol 2013; 14:169-172.
9. Malekzadeh R, Mohamadnejad M, Siavoshi F, Massarrat S. Treatment of Helicobacter pylori infection in Iran: low efficacy of recommended western regimens. Arch Iran Med 2004; 7:1-8.
10. Wroblewski LE, Peek RM, Wilson KT. Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clin Microbiol Rev 2010; 23:713-739.
11. Hasanzadeh L, Ghaznavi-Rad E, Soufian S, Farjadi V, Abtahi H. Expression and antigenic evaluation of vacA antigenic fragment of Helicobacter pylori. Iran J Basic Med Sci 2013; 16:835-840.
12. Santos AM, Lopes T, Oleastro M, Gato IV, Floch P, Benejat L, et al. Curcumin inhibits gastric inflammation induced by Helicobacter pylori infection in a mouse model. Nutrients 2015; 7:306-320.
13. Molaee N, Mosayebi G, Amozande-Nobaveh A, Soleyman MR, Abtahi H. Evolution of the Immune response against recombinant proteins (TcpA, TcpB, and FlaA) as a candidate subunit cholera vaccine. J Immunol Res 2017; 2017:2412747.
14. Yoshiyama H, Nakazawa T. Unique mechanism of Helicobacter pylori for colonizing the gastric mucus. Microbes Infect 2000; 2:55-60.
15. O'Toole PW, Lane MC, Porwollik S. Helicobacter pylori motility. Microbes Infect 2000; 2:1207-1214.
16. Clyne M, Ocroinin T, Suerbaum S, Josenhans C, Drumm B. Adherence of isogenic flagellum-negative mutants of Helicobacter pylori and Helicobacter mustelae to human and ferret gastric epithelial cells. Infect Immun 2000; 68:4335-4339.
17. Olczak AA, Olson JW, Maier RJ. Oxidative stress resistance in Helicobacter pylori. J Bacteriol 2002; 184:3186-3193.
18. Yan J, Liang SH, Mao YF, Li LW, Li SP. Construction of expression systems for flaA and flaB genes of Helicobacter pylori and determination of immunoreactivity and antigenicity of recombinant proteins. World J Gastroenterol 2003; 9:2240-2250.
19. Khan AM, Miotto O, Heiny AT, Salmon J, Srinivasan KN, Nascimento EJ, et al. A systematic bioinformatics approach for selection of epitope-based vaccine targets. Cell Immunol 2006; 244:141-147.
20. De Groot AS, Bosma A, Chinai N, Frost J, Jesdale BM, Gonzalez MA, et al. From genome to vaccine: in silico predictions, ex vivo verification. Vaccine 2001; 19:4385-4395.
21. Molaee N, Abtahi H, Mosayebi G. Expression of recombinant Streptokinase from Streptococcus pyogenes and its reaction with infected human and murine sera. Iran J Basic Med Sci 2013; 16:985-989.
22. Abbasian SS, Rad EG, Akbari N, Zolfaghari MR. Overexpression and enzymatic assessment of antigenic fragments of hyaluronidase recombinant protein from streptococcus pyogenes. Jundishapur J Microbiol 2015; 8;1-6.
23. Bonyadi M, Ghaznavi-Rad E, Mosayebi G, Rafiei M, Molaee N, Abtahi H. Development of an arCagA antigen-based assay for the detection of Helicobacter pylori in stool specimens. Jundishapur J Microbiol 2017; 10;1-5.
24. Song H, Lv X, Yang J, Liu W, Yang H, Xi T, et al. A novel chimeric flagellum fused with the multi-epitope vaccine CTB-UE prevents Helicobacter pylori-induced gastric cancer in a BALB/c mouse model. Appl Microbiol Biotechnol 2015; 99:9495-9502.
25. Guo L, Yin R, Liu K, Lv X, Li Y, Duan X, et al. Immunological features and efficacy of a multi-epitope vaccine CTB-UE against Helicobacter pylori in BALB/c mice model. Appl Microbiol Biotechnol 2014; 98:3495-3507.
26. Culver EL, Smit WL, Evans C, Sadler R, Cargill T, Makuch M, et al. No evidence to support a role for Helicobacter pylori infection and plasminogen binding protein in autoimmune pancreatitis and IgG4-related disease in a UK cohort. Pancreatology  2017; 17:395-402.
27. Tan Z, Liu W, Liu H, Li C, Zhang Y, Meng X, et al. Oral Helicobacter pylori vaccine-encapsulated acid-resistant HP55/PLGA nanoparticles promote immune protection. Eur J Pharm Biopharm 2017; 111:33-43.
28. Metzger WG, Mansouri E, Kronawitter M, Diescher S, Soerensen M, Hurwitz R, et al. Impact of vector-priming on the immunogenicity of a live recombinant Salmonella enterica serovar typhi Ty21a vaccine expressing urease A and B from Helicobacter pylori in human volunteers. Vaccine 2004; 22:2273-2277.
29. Kabir S. The current status of Helicobacter pylori vaccines: a review. Helicobacter 2007; 12:89-102.
30. Hoffelner H, Rieder G, Haas R. Helicobacter pylori vaccine development: optimisation of strategies and importance of challenging strain and animal model. Int J Med Microbiol 2008; 298:151-159.