Iranian Journal of Pathology

Comparison of Immune Response in Mice Immunized with Recombinant PreS2/S-C18-27 Protein Derived from Hepatitis B Virus with Commercial Vaccine

Document Type : Original Research

Authors

Elaheh Gholami Parizad 1
Abbas Ali Imani Fooladi 2
Hamid Sedighian 2
Elham Behzadi 3
Azar valizadeh 1
Afra Khosravi 1

1 Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran

2 Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

3 Academy of Medical Sciences of the I.R. of Iran, Tehran, Iran

https://doi.org/10.30699/ijp.2022.553785.2896
Abstract
Background & Objective: The vaccine available to prevent Hepatitis B virus disease is ineffective in 5% of people due to the use of HBsAg as a weak immunogen. In the present study, PreS2/S fused to C18-27 peptide fragment as an effective antigen and is proposed as a promising vaccine candidate compared with the conventional vaccine prescribed in the vaccination program.
Methods: After the synthesis of PreS2/S genes and C18-27 peptide fragment in pET28a, the recombinant protein was confirmed by Western blotting. The efficacy of the PreS2/S-C18-27 protein was compared with the conventional vaccine injected into five groups of rats. Finally, the cytokine level of IF-r, IL-2, IL-4, IL-10, TNF-a, IgG1, and IgG2a were measured using the ELISA method.
Results: This study showed no significant difference between the recombinant vaccine group and PBS control group in the IF-r test, but there was a significant difference between groups testing IL-2 and IL-10. In addition, the group receiving the recombinant vaccine with CPG adjuvant at a dilution of 1/10 in the IgG total test on days 14 and 45 after the first injection showed a significant difference in comparison with other groups.
Conclusion: This study showed no statistically significant difference between the recombinant protein vaccine group and the conventional vaccine group. The Th1- mediated immune responses obtained from recombinant proteins with and without CPG performed better than conventional vaccines, possibly due to the functional deficiency of the available vaccines.

Keywords

CpG 7909
Hepatitis B Core Antigen
Hepatitis B Vaccines
Hepatitis B Virus
preS2HBsAg

Subjects

  1. Nassal M. Hepatitis B viruses: reverse transcription a different way. Virus Res. 2008; 134(1-2):235-49. [DOI:10.1016/j.virusres.2007.12.024] [PMID]
  2. Locarnini S, Hatzakis A, Chen DS, Lok A. Strategies to control hepatitis B: Public policy, epidemiology, vaccine and drugs. J Hepatol. 2015;62(1 Suppl):S76-86. [DOI:10.1016/j.jhep.2015.01.018] [PMID]
  3. Brechot C. Pathogenesis of hepatitis B virus-related hepatocellular carcinoma: old and new paradigms. Gastroenterology. 2004;127(5 Suppl 1):S56-61. [DOI:10.1053/j.gastro.2004.09.016] [PMID]
  4. Tang B, Kruger WD, Chen G, Shen F, Lin WY, Mboup S, et al. Hepatitis B viremia is associated with increased risk of hepatocellular carcinoma in chronic carriers. J Med Virol. 2004;72(1):35-40. [DOI:10.1002/jmv.10559] [PMID]
  5. Lavanchy D. Worldwide epidemiology of HBV infection, disease burden, and vaccine prevention. J Clin Virol. 2005;34 Suppl 1:S1-3. [DOI:10.1016/S1386-6532(05)00384-7]
  6. Azizi F, Janqurbani M, H H. Epidemilogy and control common disease in Iran. Khosravi publication. 2004:714-41.
  7. Azizi M, Musacchio A, Pardo O, Figueroa N, Muzio V. Expression of Hepatitis B Virus Core Antigen in Native and Fusion Forms in E. coli. Iran Biomed J. 2000;4(1):37-43.
  8. Ni YH, Chang MH, Wu JF, Hsu HY, Chen HL, Chen DS. Minimization of hepatitis B infection by a 25-year universal vaccination program. J Hepatol. 2012;57(4):730-5. [DOI:10.1016/j.jhep.2012.05.021] [PMID]
  9. Chen J, Liang Z, Lu F, Fang X, Liu S, Zeng Y, et al. Toll-like receptors and cytokines/cytokine receptors polymorphisms associate with non-response to hepatitis B vaccine. Vaccine. 2011; 29(4):706-11. [DOI:10.1016/j.vaccine.2010.11.023] [PMID]
  10. Tajiri K, Shimizu Y. Unsolved problems and future perspectives of hepatitis B virus vaccination. World J Gastroenterol. 2015;21(23): 7074-83. [DOI:10.3748/wjg.v21.i23.7074] [PMID] [PMCID]
  11. Lobaina Y, Hardtke S, Wedemeyer H, Aguilar JC, Schlaphoff V. In vitro stimulation with HBV therapeutic vaccine candidate Nasvac activates B and T cells from chronic hepatitis B patients and healthy donors. Mol Immunol. 2015;63(2):320-7. [DOI:10.1016/j.molimm.2014.08.003] [PMID]
  12. Jing M, Wang J, Zhu S, Ao F, Wang L, Han T, et al. Development of a more efficient hepatitis B virus vaccine by targeting hepatitis B virus preS to dendritic cells. Vaccine. 2016;34(4):516-22. [DOI:10.1016/j.vaccine.2015.11.069] [PMID]
  13. Li J, Ge J, Ren S, Zhou T, Sun Y, Sun H, et al. Hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) combine CpG oligodeox-ynucletides as a novel therapeutic vaccine for chronic hepatitis B infection. Vaccine. 2015;33(35):4247-54. [DOI:10.1016/j.vaccine.2015.03.079] [PMID]
  14. Seeger C, Mason WS. Molecular biology of hepatitis B virus infection. Virology. 2015;479-480:672-86. [DOI:10.1016/j.virol.2015.02.031] [PMID] [PMCID]
  15. Wang S, Han Q, Zhang G, Zhang N, Li Z, Chen J, et al. CpG oligodeoxynucleotide-adjuvanted fusion peptide derived from HBcAg epitope and HIV-Tat may elicit favorable immune response in PBMCs from patients with chronic HBV infection in the immunotolerant phase. Int Immunopharmacol. 2011;11(4):406-11. [DOI:10.1016/j.intimp.2010.12.005] [PMID]
  16. Bode C, Zhao G, Steinhagen F, Kinjo T, Klinman DM. CpG DNA as a vaccine adjuvant. Expert Rev Vaccines. 2011;10(4):499-511. [DOI:10.1586/erv.10.174] [PMID] [PMCID]
  17. Cooper C, Davis H, Morris M, Efler S, Al Adhami M, Krieg A, et al. CPG 7909, an immunostimulatory TLR9 agonist oligodeoxy-nucleotide, as adjuvant to Engerix-B® HBV vaccine in healthy adults: A double-blind phase I/II study. J Clin Immunol. 2004;24(6):693-701. [DOI:10.1007/s10875-004-6244-3] [PMID]
  18. Chen X, Tang Y, Zhang Y, Zhuo M, Tang Z, Yu Y, et al. Tapasin modification on the intracellular epitope HBcAg 18-27 enhances HBV-specific CTL immune response and inhibits hepatitis B virus replication in vivo. Lab Invest. 2014;94(5): 478-90. [DOI:10.1038/labinvest.2014.6] [PMID]
  19. Wang S, Han Q, Zhang N, Chen J, Liu Z, Zhang G, et al. HBcAg18-27 epitope fused to HIV-Tat 49-57 adjuvanted with CpG ODN induces immunotherapeutic effects in transgenic mice. Immunol Lett. 2010;127(2):143-9. [DOI:10.1016/j.imlet.2009.10.011] [PMID]
  20. Shahrokhi N, Bouzari S, Jafari AAF. Priming Hepatitis B Surface (Hbsag)- and Core Antigen (Hbcag)-Specific Immune Responses by Chimeric, Hbcag with A Hbsag 'A' Determinant. Iran Biomed J. 2006;10(2):61-8.
  21. Mahboobi M, Sedighian H, Malekara E, Khalili S, Rahbar MR, Zanoos KA, et al. Harnessing an Integrative In Silico Approach to Engage Highly Immunogenic Peptides in an Antigen Design Against Epsilon Toxin (ETX) of Clostridium perfringens. Int J Pept Res Ther. 2021;27(2): 1019-26. [DOI:10.1007/s10989-020-10147-y]
  22. Mahboobi M, Sedighian H, Hedayati CH M, Bambai B, Esmaeil Soofian S, Amani J. Applying bioinformatic tools for modeling and modifying type II E. coli L-Asparginase to present a better therapeutic agent/drug for acute lymphoblastic leukemia. Intl J Cancer Manag. 2017;10(3). [DOI:10.5812/ijcm.5785]
  23. Mahboobi M, Sedighian H, Malekara E, Khalili S, Rahbar MR, Zanoos KA, et al. Harnessing an Integrative In Silico Approach to Engage Highly Immunogenic Peptides in an Antigen Design Against Epsilon Toxin (ETX) of Clostridium perfringens. Int J Pept Res Ther. 2020:1-8. [DOI:10.1007/s10989-020-10147-y]
  24. Maleki F, Sadeghifard N, Sedighian H, Bakhtiyari S, Hosseini HM, Fooladi AAI. TGFαL3-SEB fusion protein as an anticancer against ovarian cancer. Eur J Pharmacol. 2020;870:172919. [DOI:10.1016/j.ejphar.2020.172919] [PMID]
  25. Moghadam ZM, Halabian R, Sedighian H, Behzadi E, Amani J, Fooladi AAI. Designing and analyzing the structure of DT-STXB fusion protein as an anti-tumor agent: an in silico approach. Iran J Pathol. 2019;14(4):305. [DOI:10.30699/IJP.2019.101200.2004] [PMID] [PMCID]
  26. Mohseni Z, Sedighian H, Halabian R, Amani J, Behzadi E, Imani Fooladi AA. Potent in vitro antitumor activity of B-subunit of Shiga toxin conjugated to the diphtheria toxin against breast cancer. Eur J Pharmacol. 2021;899:174057. [DOI:10.1016/j.ejphar.2021.174057] [PMID]
  27. Jahangiri A, Amani J, Halabian R. In silico analyses of staphylococcal enterotoxin B as a DNA vaccine for cancer therapy. Int J Pept Res Ther. 2018;24(1):131-42. [DOI:10.1007/s10989-017-9595-3]
  28. Goleij Z, Mahmoodzadeh Hosseini H, Sedighian H, Behzadi E, Halabian R, Sorouri R, et al. Breast cancer targeted/ therapeutic with double and triple fusion Immunotoxins. J Steroid Biochem Mol Biol. 2020;200:105651. [DOI:10.1016/j.jsbmb.2020.105651] [PMID]
  29. Sedighian H, Halabian R, Amani J, Heiat M, Taheri RA, Imani Fooladi AA. Manufacturing of a novel double-function ssDNA aptamer for sensitive diagnosis and efficient neutralization of SEA. Anal Biochem. 2018;548:69-77. [DOI:10.1016/j.ab.2018.02.017] [PMID]
  30. Sedighian H, Halabian R, Amani J, Heiat M, Amin M, Fooladi AAI. Staggered Target SELEX, a novel approach to isolate non-cross-reactive aptamer for detection of SEA by apta-qPCR. J Biotechnol. 2018;286:45-55. [DOI:10.1016/j.jbiotec.2018.09.006] [PMID]
  31. Hedayati Ch M, Amani J, Sedighian H, Amin M, Salimian J, Halabian R, et al. Isolation of a new ssDNA aptamer against staphylococcal enterotoxin B based on CNBr-activated sepharose-4B affinity chromatography. J Mol Recognit. 2016;29(9):436-45. [DOI:10.1002/jmr.2542] [PMID]
  32. Jeong GU, Ahn B-Y, Jung J, Kim H, Kim T-H, Kim W, et al. A recombinant human immunoglobulin with coherent avidity to hepatitis B virus surface antigens of various viral genotypes and clinical mutants. PloS one. 2020; 15(8):e0236704. [PMID] [PMCID] [DOI:10.1371/journal.pone.0236704]
  33. Shen K, Shen L, Wang J, Jiang Z, Shen B. Understanding Amino Acid Mutations in Hepatitis B Virus Proteins for Rational Design of Vaccines and Drugs. Adv Protein Chem Struct Biol. 2015;99:131-53. [DOI:10.1016/bs.apcsb.2015.03.006] [PMID]
  34. King TH, Kemmler CB, Guo Z, Mann D, Lu Y, Coeshott C, et al. A whole recombinant yeast-based therapeutic vaccine elicits HBV X, S and Core specific T cells in mice and activates human T cells recognizing epitopes linked to viral clearance. PloS one. 2014;9(7):e101904. [PMCID] [DOI:10.1371/journal.pone.0101904] [PMID]
  35. Nishikawa K, Kimura K, Kanda Y, Sugiyama M, Kakihana K, Doki N, et al. A prospective trial of vaccine to prevent hepatitis B virus reactivation after hematopoietic stem cell transplantation. Bone Marrow Transplant. 2020;55(7):1388-98. [DOI:10.1038/s41409-020-0833-5] [PMID] [PMCID]
  36. Walayat S, Ahmed Z, Martin D, Puli S, Cashman M, Dhillon S. Recent advances in vaccination of non-responders to standard dose hepatitis B virus vaccine. World J Hepatol. 2015;7(24):2503-9. [DOI:10.4254/wjh.v7.i24.2503] [PMID] [PMCID]
  37. Schmitt S, Glebe D, Alving K, Tolle TK, Linder M, Geyer H, et al. Analysis of the pre-S2 N- and O-linked glycans of the M surface protein from human hepatitis B virus. J Biol Chem. 1999;274 (17):11945-57. [DOI:10.1074/jbc.274.17.11945] [PMID]
  38. Toita R, Kawano T, Kang JH, Murata M. Applications of human hepatitis B virus preS domain in bio- and nanotechnology. World J Gastroenterol. 2015;21(24):7400-11. [PMID] [PMCID] [DOI:10.3748/wjg.v21.i24.7400]
  39. Backes S, Jager C, Dembek CJ, Kosinska AD, Bauer T, Stephan AS, et al. Protein-prime/modified vaccinia virus Ankara vector-boost vaccination overcomes tolerance in high-antigenemic HBV-transgenic mice. Vaccine. 2016;34(7):923-32. [DOI:10.1016/j.vaccine.2015.12.060] [PMID]
  40. Kutinova L, Nemeckova S, Hamsikova E, Press M, Zavadova H, Hirsch I, et al. A recombinant vaccinia virus expressing hepatitis B virus middle surface protein. Restricted expression of HBV antigens in human diploid cells. Arch Virol. 1990;112(3-4):181-93. [DOI:10.1007/BF01323163] [PMID]
  41. Greiner VJ, Ronzon F, Larquet E, Desbat B, Esteves C, Bonvin J, et al. The structure of HBsAg particles is not modified upon their adsorption on aluminium hydroxide gel. Vaccine. 2012;30(35):5240-5. [DOI:10.1016/j.vaccine.2012.05.082] [PMID]
  42. Khodadad N, Seyedian SS, Moattari A, Biparva Haghighi S, Pirmoradi R, Abbasi S, et al. In silico functional and structural characterization of hepatitis B virus PreS/S-gene in Iranian patients infected with chronic hepatitis B virus genotype D. Heliyon. 2020;6(7):e04332. [PMID] [PMCID] [DOI:10.1016/j.heliyon.2020.e04332]
  43. Schulze A, Schieck A, Ni Y, Mier W, Urban S. Fine mapping of pre-S sequence requirements for hepatitis B virus large envelope protein-mediated receptor interaction. J Virol. 2010;84(4):1989-2000. [DOI:10.1128/JVI.01902-09] [PMID] [PMCID]
  44. Riu Garcia A. Functional Analysis of Hepatitis B Virus Variants with Mutations in the Envelope Proteins: Staats-und Universitätsbibliothek Hamburg Carl von Ossietzky; 2007.
  45. Gholizadeh M, Khanahmad H, Memarnejadian A, Aghasadeghi MR, Roohvand F, Sadat SM, et al. Design and expression of fusion protein consists of HBsAg and polyepitope of HCV as an HCV potential vaccine. Adv Biomed Res. 2015;4.
  46. Inoue T, Tanaka Y. Cross-Protection of Hepatitis B Vaccination among Different Genotypes. Vaccines. 2020;8(3):456. [PMID] [PMCID] [DOI:10.3390/vaccines8030456]
  47. Gebbing M, Bergmann T, Schulz E, Ehrhardt A. Gene therapeutic approaches to inhibit hepatitis B virus replication. World J Hepatol. 2015;7(2): 150-64. [DOI:10.4254/wjh.v7.i2.150] [PMID] [PMCID]
  48. McCluskie MJ, Davis HL. Cutting edge: CpG DNA is a potent enhancer of systemic and mucosal immune responses against hepatitis B surface antigen with intranasal administration to mice. J Immunol. 1998;161(9):4463-6.
  49. Eng NF, Bhardwaj N, Mulligan R, Diaz-Mitoma F. The potential of 1018 ISS adjuvant in hepatitis B vaccines: HEPLISAV™ review. Hum Vaccines Immunother. 2013;9(8):1661-72. [DOI:10.4161/hv.24715] [PMID] [PMCID]
  50. Liang TJ, Block TM, McMahon BJ, Ghany MG, Urban S, Guo JT, et al. Present and future therapies of hepatitis B: From discovery to cure. Hepatology. 2015;62(6):1893-908. [DOI:10.1002/hep.28025] [PMID] [PMCID]
  51. Pollicino T, Cacciola I, Saffioti F, Raimondo G. Hepatitis B virus PreS/S gene variants: pathobiology and clinical implications. J Hepatol. 2014;61(2):408-17. [DOI:10.1016/j.jhep.2014.04.041] [PMID]
  52. Julithe R, Abou-Jaoude G, Sureau C. Modification of the hepatitis B virus envelope protein glycosylation pattern interferes with secretion of viral particles, infectivity, and susceptibility to neutralizing antibodies. J Virol. 2014;88(16):9049-59. [DOI:10.1128/JVI.01161-14] [PMID] [PMCID]
  53. Schmitt S, Glebe D, Alving K, Tolle TK, Linder M, Geyer H, et al. Analysis of the Pre-S2 N- and O-Linked Glycans of the M Surface Protein from Human Hepatitis B Virus*. J Biol Chem. 1999; 274(17):11945-57. [DOI:10.1074/jbc.274.17.11945] [PMID]
  54. Gerlich WH. Prophylactic vaccination against hepatitis B: achievements, challenges and perspectives. Med Microbiol Immunol. 2015;204 (1):39-55. [DOI:10.1007/s00430-014-0373-y] [PMID]

 

Iranian Journal of Pathology
Volume 17, Issue 4
Autumn 2022
Pages 448-460

  • Receive Date 22 May 2022
  • Revise Date 05 July 2022
  • Accept Date 13 July 2022

Home

Guide for Authors

Submit Manuscript

Contact Us