Recombinant hemagglutinin of swine H1N1 influenza virus expression in the insect cells: Formulation in Montanide ISA71 adjuvant and the potency studies

Document Type : Original Article

Authors

1 Department of Microbiology, Faculty of Advanced science and Technology, Tehran Medical sciences Islamic Azad University, Tehran, Iran

2 Department of Research and Development, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

3 Production and Research Complex, Pasteur Institute of Iran, Karaj, Iran

4 Advanced Therapy Medicinal Product Department, Breast Cancer Research Center, Motamed Cancer Institute, Academic Center for Education, Culture and Research, Tehran, Iran

5 Recombinant Vaccine Research Center, Tehran University of Medical Sciences, Tehran, Iran

6 Immunotherapy Group, Pharmaceutical Sciences Research Center (PSRC), The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences, Tehran, Iran

7 Department of Immunology, Pasteur Institute of Iran, Tehran, Iran

Abstract

Objective(s): Influenza is a highly contagious disease, which affects the respiratory system and seasonal influenza is common throughout the world. Influenza vaccination is an effective way to reduce the risk of death and hospitalization. This study aims at the expression of swine recombinant hemagglutinin protein in the baculovirus expression system and it offers a comparison of the immunologic parameters with the commercial vaccine. 
Materials and Methods: The HA gene from the swine H1N1 strain of the Influenza virus was cloned into the Bac-To-Bac expression system in pFastBAC HTA vector and was transformed into Escherichia coli TOP10 strain. After the confirmation, the vector was transfected into the SF9 insect cell line. The recombinant HA was evaluated by SDS-PAGE and western blot. After formulation in Montanide ISA71 adjuvant, the immunization test was performed comparatively with Alum adjuvant, commercial vaccine in four groups of BALB/c mice, of which one group was control without any vaccination. Two weeks after the last immunization, the antibody response was assessed with HI assay, and experimental mice were challenged with mouse-adapted Influenza A/PR8/34 (H1N1) virus through nasal inhalation. 
Results: The immunoassay results revealed that the candidate vaccine induced the antibody response as the commercial one did but it did not significantly reduce the mortality rate, body loss, and severe fever. 
Conclusion: To summarize, the results showed that the recombinant protein with the MontanideTM ISA- 71 adjuvant developed a more appropriate level of immunity than Alum adjuvant, so it might be used as a safe and reliable vaccine against H1N1 virus for further research.

Keywords


1. Harding AT, Heaton NS. Efforts to improve the seasonal influenza vaccine. Vaccines 2018; 6:19-31.
2. Wright P, Neumann G, Kawaoka Y. Chapter 48 Orthomyxoviruses. Fields Virology Fifth Edition Philadelphia, PA, USA: Lippincott Williams & Amp; Wilkins 2007:1691-1740.
3. Cox NJ, Tamblyn SE, Tam T. Influenza pandemic planning. Vaccine 2003; 21:1801-1803.
4. Kilbourne ED. Influenza pandemics of the 20th century. Emerg Infect Dis 2006; 12:9-14.
5. Gendon I. Influenza pandemic: hypotheses and facts. Zh Mikrobiol Epidemiol Immunobiol 2008; 5:109-118.
6. Saunders-Hastings PR, Krewski D. Reviewing the history of pandemic influenza: understanding patterns of emergence and transmission. Pathogens 2016; 5:66-85.
7.Kitikoon P, Vincent AL, Janke BH, Erickson B, Strait EL, Yu S, et al. Swine influenza matrix 2 (M2) protein contributes to protection against infection with different H1 swine influenza virus (SIV) isolates. Vaccine 2009; 28:523-531.
8. Frolov VG, Seid Jr RC, Odutayo O, Al‐Khalili M, Yu J, Frolova OY, et al. Transcutaneous delivery and thermostability of a dry trivalent inactivated influenza vaccine patch. Influenza Other Respir Viruses 2008; 2:53-60.
9. Mameli C, Cocchi I, Fumagalli M, Zuccotti G. Influenza vaccination: effectiveness, indications, and limits in the pediatric population. Front Pediatr 2019; 7:317-325.
10. Maeda Y, Horimoto T, Kawaoka Y. Classification and genome structure of influenza virus. Nippon Rinsho 2003; 61:1886-1891.
11.Pavlovic J, Schultz J, Hefti H, Schuh T, Mölling K. DNA vaccination against la crosse virus. Intervirology 2000; 43:312-321.
12. Rockman S, Laurie KL, Parkes S, Wheatley A, Barr IG. New technologies for influenza vaccines. Microorganisms 2020; 8:1745-1765.
13.Eleftheratos S, Spearpoint P, Ortore G, Kolocouris A, Martinelli A, Martin S, et al. Interaction of aminoadamantane derivatives with the influenza A virus M2 channel-docking using a pore blocking model. Bioorganic Med Chem Lett 2010; 20:4182-4187.
14. Ruigrok RW, Barge A, Durrer P, Brunner J, Ma K, Whittaker GR, et al. Membrane interaction of influenza virus M1 protein. Virol 2000; 267:289-298.
15. Demoly P, Dhivert-Donnadieu H, Bousquet J. Vaccination with allergens in children. Allerg Immunol 2000; 32:397-401.
16. McNeil MM, DeStefano F. Vaccine-associated hypersensitivity. J Allergy Clin Immunol 2018; 141:463-472.
17. Perrie Y, Mohammed AR, Kirby DJ, McNeil SE, Bramwell VW. Vaccine adjuvant systems: enhancing the efficacy of sub-unit protein antigens. Int J Pharm 2008; 364:272-280.
18. Girard MP, Katz JM, Pervikov Y, Hombach J, Tam JS. Report of the 7th meeting on evaluation of pandemic influenza vaccines in clinical trials, world health organization, Geneva, 17–18 February 2011. Vaccine 2011; 29:7579-7586.
19. Shokouhi H, Zolfaghari MR, Farahmand B, Tabatabaeian M, Taheri N, Fotouhi F, et al. Immunological assessment of three tandem repeat of influenza virus M2 extracellular domain with adjuvant in balb/c mice model. J ARAK Uni Med Sci 2016; 19:32-43.
20. Loghmani A, Delirezh N, Moattari A. Development of MF-59 adjuvant using chitosan derivatives to influenza vaccine. J Biomed Sci 2020; 10:2261-2271.
21. Chambers AC, Aksular M, Graves LP, Irons SL, Possee RD, King LA et al. Overview of the baculovirus expression system. Curr Protoc Protein Sci 2018; 91:1-6.
22. Yang Z-Y, Wei C-J, Kong W-P, Wu L, Xu L, Smith DF, et al. Immunization by avian H5 influenza hemagglutinin mutants with altered receptor binding specificity. Sci 2007; 317:825-828.
23. Bouvier NM, Lowen AC. Animal models for influenza virus pathogenesis and transmission. Viruses 2010; 2:1530-1563.
24. Killian ML. Hemagglutination assay for influenza virus. Method Mol Biol 2020; 2123:3-10.
25. Chen J-R, Liu Y-M, Tseng Y-C, Ma C. Better influenza vaccines: an industry perspective. J Biomed Sci 2020; 27:1-11.
26. Yang S, Lee J-Y, Lee JS, Mitchell WP, Oh H-B, Kang C, et al. Influenza sequence and epitope database. Nucleic Acids Res 2009; 37:423-430.
27. Wang K, Holtz KM, Anderson K, Chubet R, Mahmoud W, Cox MM, et al. Expression and purification of an influenza hemagglutinin—one step closer to a recombinant protein-based influenza vaccine. Vaccine  2006; 24:2176-2185.
28. Hoffmann E, Lipatov AS, Webby RJ, Govorkova EA, Webster RG. Role of specific hemagglutinin amino acids in the immunogenicity and protection of H5N1 influenza virus vaccines. Proc Natl Acad Sci 2005; 102:12915-12920.
29. Treanor JJ, Essink B, Hull S, Reed S, Izikson R, Patriarca P, et al. Evaluation of safety and immunogenicity of recombinant influenza hemagglutinin (H5/Indonesia/05/2005) formulated with and without a stable oil-in-water emulsion containing glucopyranosyl-lipid A (SE+ GLA) adjuvant. Vaccine 2013; 31:5760-5765.
30. Ophorst OJ, Radošević K, Klap JM, Sijtsma J, Gillissen G, Mintardjo R, et al. Increased immunogenicity of recombinant Ad35-based malaria vaccine through formulation with aluminium phosphate adjuvant. Vaccine 2007; 25:6501-6510.
31. Baumert TF, Ito S, Wong DT, Liang TJ. Hepatitis C virus structural proteins assemble into viruslike particles in insect cells. J Virol 1998; 72:3827-3836.
32. Hu YC, Luo YL, Ji WT, Chulu JL, Chang PC, Shieh H, et al. Dual expression of the HA protein of H5N2 avian influenza virus in a baculovirus system. J Virol Methods 2006; 135:43-48.
33. Krammer F, Fouchier RA, Eichelberger MC, Webby RJ, Shaw-Saliba K, Wan H, et al. Naction! How can neuraminidase-based immunity contribute to better influenza virus vaccines?  MBio 2018; 3:9-21.
34. Luckow VA, Lee SC, Barry GF, Olins P. Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J Virol 1993; 67:4566-4579.
35. Matsuda T, Tanijima T, Hirose A, Masumi-Koizumi K, Katsuda T, Yamaji H, et al. Production of influenza virus-like particles using recombinant insect cells. Biochem Eng J 2020; 163:107757-107764.
36. Feshchenko E, Rhodes DG, Felberbaum R, McPherson C, Rininger JA, Post P, et al. Pandemic influenza vaccine: characterization of A/California/07/2009 (H1N1) recombinant hemagglutinin protein and insights into H1N1 antigen stability. BMC Biotechnol 2012; 12:1-13.
37. Ebrahimi SM, Tebianian M, Aghaiypour K, Nili H, Mirjalili A. Prokaryotic expression and characterization of avian influenza A virus M2 gene as a candidate for universal recombinant vaccine against influenza A subtypes; specially H5N1 and H9N2. Mol Biol Rep 2010; 37:2909-2914.
38. Bungener L, Geeraedts F, Ter Veer W, Medema J, Wilschut J, Huckriede A, et al. Alum boosts TH2-type antibody responses to whole-inactivated virus influenza vaccine in mice but does not confer superior protection. Vaccine 2008; 26:2350-2359.
39. Jafari M, Moghaddam Pour M, Taghizadeh M, Masoudi S, Bayat Z. Comparative assessment of humoral immune responses of aluminum hydroxide and oil-emulsion adjuvants in Influenza (H9N2) and newcastle inactive vaccines to chickens. Artif Cells  Nanomed Biotechnol 2017; 45:84-89.
40. Miller LH, Saul A, Mahanty S. Revisiting Freund’s incomplete adjuvant for vaccines in the developing world. Trends Parasitol 2005; 21:412-414.