Immunogenicity Enhancement of Influenza Virus Stalk Domain Using Leishmania Major Heat Shock Protein, One Step Closer to Universal Vaccine

Document Type : Original Article (s)

Authors

1 Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Assistant Professor, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran

3 Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran

Abstract

Background: Influenza viruses cause acute respiratory infections in humans, which can often be accompanied by pandemics with varying rates of illness and even a fetal end. Current vaccines target the variable globular part of hemagglutinin (HA); so, their effectiveness is limited and they need perpetual updating. To prepare universal vaccine, scientists' effort is focused on conserved antigenic parts of influenza virus such as hemagglutinin stalk domain which is more conserved compare to its globular head. The aim of this study was evaluating the immunogenicity of influenza A (H1N1) HA2 peptide in mouse model.Methods: To prepare HA2 protein, the expression vector encoding the gene of interest (PET-28a/HA2), was transformed into Escherichia coli BL21-DE3 competent cells. Recombinant HA2 protein was extracted and purified using Ni-TED columns under denaturing conditions, refolded and desalted via step-wise dialysis. The HA2 concentration was determined via using Bradford protein assay. Six-week-old BALB/c mice in different groups were immunized intradermally with HA2 alone or supplemented with Freund or Lm.HSP70221-604 as adjuvants. Two groups were injected with HSP70 and phosphate buffered saline (PBS) as negative controls. Humeral immunity in vaccinated mice was measured using the enzyme-linked immunosorbent assay (ELISA); and finally, mice were challenged with one lethal dose (LD90) of PR8 virus.Findings: The protein expression, extraction and purification stages were confirmed using sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) electrophoresis. The highest antibody response was measured in mice vaccinated with HA2-Freund, as expected. Specific antibody response in group received HA2-HSP70 was of higher titer than HA2 alone. More over HA2-HSP70 group surprisingly showed higher level of protection against lethal H1N1 (PR8) challenge compared to other groups.Conclusions: It has been proved that HSP70 can induce specific response and innate immunity via stimulating CD8+ T-Cells, CD40, TLR2, TLR4 and cytokine secretion. Our results showed that HA2 alone or supplemented with HSP70 protected mice against lethal influenza challenge and could be considered as a universal vaccine.

Keywords

References

  1. Lambert LC, Fauci AS. Influenza vaccines for the future. N Engl J Med 2010; 363(21): 2036-44.
  2. Hara K, Shiota M, Kido H, Ohtsu Y, Kashiwagi T, Iwahashi J, et al. Influenza virus RNA polymerase PA subunit is a novel serine protease with Ser624 at the active site. Genes Cells 2001; 6(2): 87-97.
  3. Okuda K, Ihata A, Watabe S, Okada E, Yamakawa T, Hamajima K, et al. Protective immunity against influenza A virus induced by immunization with DNA plasmid containing influenza M gene. Vaccine 2001; 19(27): 3681-91.
  4. Kim CS, Epand RF, Leikina E, Epand RM, Chernomordik LV. The final conformation of the complete ectodomain of the HA2 subunit of influenza hemagglutinin can by itself drive low pH-dependent fusion. J Biol Chem 2011; 286(15): 13226-34.
  5. Kim JI, Lee I, Park S, Park MS. Surface glycoproteins determine the feature of the 2009 pandemic H1N1 virus. BMB Rep 2012; 45(11): 653-8.
  6. Reichert T, Chowell G, Nishiura H, Christensen RA, McCullers JA. Does Glycosylation as a modifier of Original Antigenic Sin explain the case age distribution and unusual toxicity in pandemic novel H1N1 influenza? BMC Infect Dis 2010; 10: 5.
  7. Peiris JS, Tu WW, Yen HL. A novel H1N1 virus causes the first pandemic of the 21st century. Eur J Immunol 2009; 39(11): 2946-54.
  8. Fujiyoshi Y, Kume NP, Sakata K, Sato SB. Fine structure of influenza A virus observed by electron cryo-microscopy. EMBO J 1994; 13(2): 318-26.
  9. Cox NJ, Subbarao K. Global epidemiology of influenza: past and present. Annu Rev Med 2000; 51: 407-21.
  10. Hilleman MR. Realities and enigmas of human viral influenza: pathogenesis, epidemiology and control. Vaccine 2002; 20(25-26): 3068-87.
  11. Ebrahimi SM, Tebianian M. Influenza A viruses: why focusing on M2e-based universal vaccines. Virus Genes 2011; 42(1): 1-8.
  12. Black RA, Rota PA, Gorodkova N, Klenk HD, Kendal AP. Antibody response to the M2 protein of influenza A virus expressed in insect cells. J Gen Virol 1993; 74 ( Pt 1): 143-6.
  13. Nicholson KG, Wood JM, Zambon M. Influenza. Lancet 2003; 362(9397): 1733-45.
  14. Stanekova Z, Vareckova E. Conserved epitopes of influenza A virus inducing protective immunity and their prospects for universal vaccine development. Virol J 2010; 7: 351.
  15. Fouchier RA, Munster V, Wallensten A, Bestebroer TM, Herfst S, Smith D, et al. Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls. J Virol 2005; 79(5): 2814-22.
  16. Stanekova Z, Mucha V, Sladkova T, Blaskovicova H, Kostolansky F, Vareckova E. Epitope specificity of anti-HA2 antibodies induced in humans during influenza infection. Influenza Other Respir Viruses 2012; 6(6): 389-95.
  17. Vareckova E, Mucha V, Kostolansky F. HA2 glycopolypeptide of influenza A virus and antiviral immunity. Acta Virol 2013; 57(2): 247-56.
  18. Horvath A, Toth GK, Gogolak P, Nagy Z, Kurucz I, Pecht I, et al. A hemagglutinin-based multipeptide construct elicits enhanced protective immune response in mice against influenza A virus infection. Immunol Lett 1998; 60(2-3): 127-36.
  19. Okuno Y, Matsumoto K, Isegawa Y, Ueda S. Protection against the mouse-adapted A/FM/1/47 strain of influenza A virus in mice by a monoclonal antibody with cross-neutralizing activity among H1 and H2 strains. J Virol 1994; 68(1): 517-20.
  20. Sui J, Hwang WC, Perez S, Wei G, Aird D, Chen LM, et al. Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses. Nat Struct Mol Biol 2009; 16(3): 265-73.
  21. Kostolansky F, Mucha V, Slovakova R, Vareckova E. Natural influenza A virus infection of mice elicits strong antibody response to HA2 glycopolypeptide. Acta Virol 2002; 46(4): 229-36.
  22. Steel J, Lowen AC, Wang TT, Yondola M, Gao Q, Haye K, et al. Influenza virus vaccine based on the conserved hemagglutinin stalk domain. MBio 2010; 1(1): e00018-10.
  23. To KK, Zhang AJ, Chan AS, Li C, Cai JP, Lau CC, et al. Recombinant influenza A virus hemagglutinin HA2 subunit protects mice against influenza A(H7N9) virus infection. Arch Virol 2015; 160(3): 777-86.
  24. Kang SM, Song JM, Compans RW. Novel vaccines against influenza viruses. Virus Res 2011; 162(1-2): 31-8.
  25. Awate S, Babiuk LA, Mutwiri G. Mechanisms of action of adjuvants. Front Immunol 2013; 4: 114.
  26. Rafati S, Gholami E, Hassani N, Ghaemimanesh F, Taslimi Y, Taheri T, et al. Leishmania major heat shock protein 70 (HSP70) is not protective in murine models of cutaneous leishmaniasis and stimulates strong humoral responses in cutaneous and visceral leishmaniasis patients. Vaccine 2007; 25(21): 4159-69.
  27. Foumani M, Asadpour L, Azizi Saraji A, Sharifat Salmani A, Aghasadeghi MR. Adjuvants and their mechanisms of action. J Ardabil Univ Med Sci 2012; 12(3): 276-91. [In Persian].
  28. Janulikova J, Stanekova Z, Mucha V, Kostolansky F, Vareckova E. Two distinct regions of HA2 glycopolypeptide of influenza virus hemagglutinin elicit cross-protective immunity against influenza. Acta Virol 2012; 56(3): 169-76.
Journal of Isfahan Medical School
Volume 33, Issue 349 - Serial Number 349
July and August 2015
Pages 1475-1486

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History

  • Receive Date: 16 June 2015
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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