Immunogenicity assessment of Hepatitis A-VP1 and Hepatitis B surface antigen (HBsAg) fusion protein: a novel bivalent vaccine candidate
,
Abstract
Background and Objectives: Subunit vaccines have the privilege of utilizing immunogenic parts of the variable viruses. The current preventive vaccines against Hepatitis A are based on live-attenuated virus or wild-type growth in cell culture, which is a time-consuming and costly procedure. Thus, the investigation of immunogenic Hepatitis A Virus (HAV) regions seems to be a rational priority. We aimed to evaluate a novel chimeric protein composed of truncated HAV-VP1 and Hepatitis B surface antigen (HBsAg) as a bivalent vaccine candidate in BALB/c mice.
Materials and Methods: The HAV-VP1 (amino acids 99 to 259) and HBsAg fusion protein were applied as a bivalent vaccine in combination with adjuvants. The purified protein was administered through different regimens via subcutaneous injection. Two weeks following the final immunization, serum samples were gathered to assess the humoral responses. Moreover, splenocytes were investigated and assessed for IL-5 and IFN-γ secretion.
Results: The immunized mice with recombinant truncated HAV-VP1-AAY-HBsAg showed a significant immune response, especially in combination with the M720 adjuvant. Humoral immune response results indicated Th1 switching by IgG2a and IgG2b dominancy. Moreover, IFN-γ secretion reached the highest rate in the truncated HAV-VP1-AAY-HBsAg+M720 recipients (p<0.0001).
Conclusion: The HAV-VP1-AAY-HBsAg protein subunit vaccine could help the immune system fight HAV and HBV by stimulating both the humoral and cellular immune systems. The formula proposed in this study has the potential to produce an endemic vaccine based on the circulating HAV viruses in Iran.
1. Castaneda D, Gonzalez AJ, Alomari M, Tandon K, Zervos XB. From hepatitis A to E: A critical review of viral hepatitis. World J Gastroenterol 2021; 27: 1691-1715.
2. Bhadoria AS, Khwairakpam G, Grover GS, Pathak VK, Pandey P, Gupta R. Viral hepatitis as a public health concern: a narrative review about the current scenario and the way forward. Cureus 2022; 14(2): e21907.
3. Migueres M, Lhomme S, Izopet J. Hepatitis A: Epidemiology, high-risk groups, prevention and research on antiviral treatment. Viruses 2021; 13: 1900.
4. Sayed IM. Dual infection of hepatitis A virus and hepatitis E virus—What is known? Viruses 2023; 15: 298.
5. Wang X, Ren J, Gao Q, Hu Z, Sun Y, Li X, et al. Hepatitis A virus and the origins of picornaviruses. Nature 2015; 517: 85-88.
6. WHO position paper on hepatitis A vaccines-June 2012. Wkly Epidemiol Rec 2012; 87: 261-276.
7. Rein DB, Stevens G, Flaxman A, Wittenborn JS, Timothy N, Wiktor SZ, et al. P703 The global burden of hepatitis a virus in 1990 and 2005. J Hepatol 2014; 1: S303.
8. Hannan M, Jabalameli L, Aghasadeghi MR, Harzandi N, Sadat SM. Expression and evaluation of a novel HAV-VP1 and HBS-Ag Fusion protein for Potential applications in Immunization and diagnosis. J Med Microbiol Infect Dis 2023; 11: 128-135.
9. Guzman-Holst A, Luna-Casas G, Burguete Garcia A, Madrid-Marina V, Cervantes-Apolinar MY, Andani A, et al. Burden of disease and associated complications of hepatitis a in children and adults in Mexico: A retrospective database study. PLoS One 2022; 17(5): e0268469.
10. Larijani MS, Ramezani A, Sadat SM. Updated studies on the development of HIV therapeutic vaccine. Curr HIV Res 2019; 17: 75-84.
11. Larijani MS, Sadat SM, Bolhassani A, Khodaie A, Pouriayevali MH, Ramezani A. HIV-1 p24-nef DNA vaccine plus protein boost expands T-Cell responses in BALB/c. Curr Drug Deliv 2021; 18: 1014-1021.
12. Pollard AJ, Bijker EM. A guide to vaccinology: from basic principles to new developments. Nat Rev Immunol 2021; 21: 83-100.
13. Nelson NP, Link-Gelles R, Hofmeister MG, Romero JR, Moore KL, Ward JW, et al. Update: recommendations of the Advisory Committee on Immunization Practices for use of hepatitis A vaccine for postexposure prophylaxis and for preexposure prophylaxis for international travel. MMWR Morb Mortal Wkly Rep 2018; 67: 1216-1220.
14. Victor JC, Monto AS, Surdina TY, Suleimenova SZ, Vaughan G, Nainan OV, et al. Hepatitis A vaccine versus immune globulin for postexposure prophylaxis. N Engl J Med 2007; 357: 1685-1694.
15. Nguyen MH, Wong G, Gane E, Kao JH, Dusheiko G. Hepatitis B virus: advances in prevention, diagnosis, and therapy. Clin Microbiol Rev 2020; 33(2): e00046-19.
16. Van Damme E, Vanhove J, Severyn B, Verschueren L, Pauwels F. The hepatitis b virus interactome: A comprehensive overview. Front Microbiol 2021; 12: 724877.
17. Ginzberg D, Wong RJ, Gish R. Global HBV burden: guesstimates and facts. Hepatol Int 2018; 12: 315-329.
18. Yuen MF, Chen DS, Dusheiko GM, Janssen HLA, Lau DTY, Locarnini SA, et al. Hepatitis B virus infection. Nat Rev Dis Primers 2018; 4: 18035.
19. Alberts CJ, Clifford GM, Georges D, Negro F, Lesi OA, Hutin YJ, et al. Worldwide prevalence of hepatitis B virus and hepatitis C virus among patients with cirrhosis at country, region, and global levels: a systematic review. Lancet Gastroenterol Hepatol 2022; 7: 724-735.
20. Ho JK, Jeevan-Raj B, Netter HJ. Hepatitis B virus (HBV) subviral particles as protective vaccines and vaccine platforms. Viruses 2020; 12: 126.
21. Anutebeh EN, Tatah L, Feteh VF, Aroke D, Assob JCN, Choukem SP. Immune response to hepatitis B vaccine following complete immunization of children attending two regional hospitals in the Southwest region of Cameroon: a cross sectional study. BMC Infect Dis 2021; 21: 1205.
22. Hill HA, Elam-Evans LD, Yankey D, Singleton JA, Kolasa M. National, state, and selected local area vaccination coverage among children aged 19-35 months-United States, 2014. MMWR Morb Mortal Wkly Rep 2015; 64: 889-896.
23. Larijani MS, Sadat SM, Bolhassani A, Pouriayevali MH, Bahramali G, Ramezani A. In silico design and immunologic evaluation of HIV-1 p24-Nef fusion protein to approach a therapeutic vaccine candidate. Curr HIV Res 2018; 16: 322-337.
24. Sadat Larijani M, Ramezani A, Mashhadi Abolghasem Shirazi M, Bolhassani A, Pouriayevali MH, Shahbazi S, et al. Evaluation of transduced dendritic cells expressing HIV-1 p24-Nef antigens in HIV-specific cytotoxic T cells induction as a therapeutic candidate vaccine. Virus Res 2021; 298: 198403.
25. Beran J. Bivalent inactivated hepatitis A and recombinant hepatitis B vaccine. Expert Rev Vaccines 2007; 6: 891-902.
26. Senger T, Schädlich L, Gissmann L, Müller M. Enhanced papillomavirus-like particle production in insect cells. Virology 2009; 388: 344-353.
27. Nain A, Kumar M, Banerjee M. Oligomers of hepatitis A virus (HAV) capsid protein VP1 generated in a heterologous expression system. Microb Cell Fact 2022; 21: 53.
28. Feng R, Wang L, Shi D, Zheng B, Zhang L, Hou H. Structural basis for neutralization of an anicteric hepatitis associated echovirus by a potent neutralizing antibody. Cell Discov 2021; 7: 35.
29. Ahmadi N, Aghasadeghi M, Hamidi-Fard M, Motevalli F, Bahramali G. Reverse vaccinology and immunoinformatic approach for designing a bivalent vaccine candidate against hepatitis A and hepatitis B viruses. Mol Biotechnol 2024; 66: 2362-2380.
30. Ayyagari VS, T C V, K AP, Srirama K. Design of a multi-epitope-based vaccine targeting M-protein of SARS-CoV2: an immunoinformatics approach. J Biomol Struct Dyn 2022; 40: 2963-2977.
31. Chao P, Zhang X, Zhang L, Yang A, Wang Y, Chen X. Proteomics-based vaccine targets annotation and design of multi-epitope vaccine against antibiotic-resistant Streptococcus gallolyticus. Sci Rep 2024; 14: 4836.
32. Stuurman AL, Marano C, Bunge EM, De Moerlooze L, Shouval D. Impact of universal mass vaccination with monovalent inactivated hepatitis A vaccines–A systematic review. Hum Vaccin Immunother 2017; 13: 724-736.
33. Lemon SM, Ott JJ, Van Damme P, Shouval D. Type A viral hepatitis: A summary and update on the molecular virology, epidemiology, pathogenesis and prevention. J Hepatol 2017; S0168-8278(17)32278-X.
34. Nain A, Kumar M, Banerjee M. Oligomers of hepatitis A virus (HAV) capsid protein VP1 generated in a heterologous expression system. Microb Cell Fact 2022; 21: 53.
35. Larijani MS, Pouriayevali MH, Sadat SM, Ramezani A. Production of recombinant HIV-1 p24-Nef protein in two forms as potential candidate vaccines in three vehicles. Curr Drug Deliv 2020; 17: 387-395.
36. Ai J, Zhang H, Zhang Q, Zhang Y, Lin K, Fu Z, et al. Recombinant protein subunit vaccine booster following two-dose inactivated vaccines dramatically enhanced anti-RBD responses and neutralizing titers against SARS-CoV-2 and Variants of Concern. Cell Res 2022; 32: 103-106.
37. Beaumont E, Patient R, Hourioux C, Dimier-Poisson I, Roingeard P. Chimeric hepatitis B virus/hepatitis C virus envelope proteins elicit broadly neutralizing antibodies and constitute a potential bivalent prophylactic vaccine. Hepatology 2013; 57: 1303-1313.
38. Van Damme P, Van Herck K. A review of the efficacy, immunogenicity and tolerability of a combined hepatitis A and B vaccine. Expert Rev Vaccines 2004; 3: 249-267.
39. Jang KO, Park JH, Lee HH, Chung DK, Kim W, Chung IS. Expression and immunogenic analysis of recombinant polypeptides derived from capsid protein VP1 for developing subunit vaccine material against hepatitis A virus. Protein Expr Purif 2014; 100: 1-9.
40. Banerjee K, Klasse PJ, Sanders RW, Pereyra F, Michael E, Lu M, et al. IgG subclass profiles in infected HIV type 1 controllers and chronic progressors and in uninfected recipients of Env vaccines. AIDS Res Hum Retroviruses 2010; 26: 445-458.
41. Lee JM, Lee HH, Hwang-Bo J, Shon DH, Kim W, Chung IS. Expression and immunogenicity of recombinant polypeptide VP1 of human hepatitis A virus in stably transformed fruitfly (Drosophila melanogaster) Schneider 2 cells. Biotechnol Appl Biochem 2009; 53: 101-109.
42. Kang S, Brown HM, Hwang S. Direct antiviral mechanisms of interferon-gamma. Immune Netw 2018; 18(5): e33.
43. Shin EC, Sung PS, Park SH. Immune responses and immunopathology in acute and chronic viral hepatitis. Nat Rev Immunol 2016; 16: 509-523.
44. Chung HY, Lee HH, Kim KI, Chung HY, Hwang-Bo J, Park JH, et al. Expression of a recombinant chimeric protein of hepatitis A virus VP1-Fc using a replicating vector based on Beet curly top virus in tobacco leaves and its immunogenicity in mice. Plant Cell Rep 2011; 30: 1513-1521.
45. Mutiso JM, Macharia JC, Kariuki TM, Gicheru MM. Montanide ISA 720 is more effective than BCG as an adjuvant for Leishmania killed vaccine in BALB/c mice. Int J Integ Biol 2009; 7: 107-116.
46. Marques RF, de Melo FM, Novais JT, Soares IS, Bargieri DY, Gimenez AM. Immune system modulation by the adjuvants poly (I: C) and montanide ISA 720. Front Immunol 2022; 13: 910022.
47. Aucouturier J, Dupuis L, Deville S, Ascarateil S, Ganne V. Montanide ISA 720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines. Expert Rev Vaccines 2002; 1: 111-118.
48. Marques RF, de Melo FM, Novais JT, Soares IS, Bargieri DY, Gimenez AM. Immune system modulation by the adjuvants poly (I: C) and montanide ISA 720. Front Immunol 2022; 13: 910022.
2. Bhadoria AS, Khwairakpam G, Grover GS, Pathak VK, Pandey P, Gupta R. Viral hepatitis as a public health concern: a narrative review about the current scenario and the way forward. Cureus 2022; 14(2): e21907.
3. Migueres M, Lhomme S, Izopet J. Hepatitis A: Epidemiology, high-risk groups, prevention and research on antiviral treatment. Viruses 2021; 13: 1900.
4. Sayed IM. Dual infection of hepatitis A virus and hepatitis E virus—What is known? Viruses 2023; 15: 298.
5. Wang X, Ren J, Gao Q, Hu Z, Sun Y, Li X, et al. Hepatitis A virus and the origins of picornaviruses. Nature 2015; 517: 85-88.
6. WHO position paper on hepatitis A vaccines-June 2012. Wkly Epidemiol Rec 2012; 87: 261-276.
7. Rein DB, Stevens G, Flaxman A, Wittenborn JS, Timothy N, Wiktor SZ, et al. P703 The global burden of hepatitis a virus in 1990 and 2005. J Hepatol 2014; 1: S303.
8. Hannan M, Jabalameli L, Aghasadeghi MR, Harzandi N, Sadat SM. Expression and evaluation of a novel HAV-VP1 and HBS-Ag Fusion protein for Potential applications in Immunization and diagnosis. J Med Microbiol Infect Dis 2023; 11: 128-135.
9. Guzman-Holst A, Luna-Casas G, Burguete Garcia A, Madrid-Marina V, Cervantes-Apolinar MY, Andani A, et al. Burden of disease and associated complications of hepatitis a in children and adults in Mexico: A retrospective database study. PLoS One 2022; 17(5): e0268469.
10. Larijani MS, Ramezani A, Sadat SM. Updated studies on the development of HIV therapeutic vaccine. Curr HIV Res 2019; 17: 75-84.
11. Larijani MS, Sadat SM, Bolhassani A, Khodaie A, Pouriayevali MH, Ramezani A. HIV-1 p24-nef DNA vaccine plus protein boost expands T-Cell responses in BALB/c. Curr Drug Deliv 2021; 18: 1014-1021.
12. Pollard AJ, Bijker EM. A guide to vaccinology: from basic principles to new developments. Nat Rev Immunol 2021; 21: 83-100.
13. Nelson NP, Link-Gelles R, Hofmeister MG, Romero JR, Moore KL, Ward JW, et al. Update: recommendations of the Advisory Committee on Immunization Practices for use of hepatitis A vaccine for postexposure prophylaxis and for preexposure prophylaxis for international travel. MMWR Morb Mortal Wkly Rep 2018; 67: 1216-1220.
14. Victor JC, Monto AS, Surdina TY, Suleimenova SZ, Vaughan G, Nainan OV, et al. Hepatitis A vaccine versus immune globulin for postexposure prophylaxis. N Engl J Med 2007; 357: 1685-1694.
15. Nguyen MH, Wong G, Gane E, Kao JH, Dusheiko G. Hepatitis B virus: advances in prevention, diagnosis, and therapy. Clin Microbiol Rev 2020; 33(2): e00046-19.
16. Van Damme E, Vanhove J, Severyn B, Verschueren L, Pauwels F. The hepatitis b virus interactome: A comprehensive overview. Front Microbiol 2021; 12: 724877.
17. Ginzberg D, Wong RJ, Gish R. Global HBV burden: guesstimates and facts. Hepatol Int 2018; 12: 315-329.
18. Yuen MF, Chen DS, Dusheiko GM, Janssen HLA, Lau DTY, Locarnini SA, et al. Hepatitis B virus infection. Nat Rev Dis Primers 2018; 4: 18035.
19. Alberts CJ, Clifford GM, Georges D, Negro F, Lesi OA, Hutin YJ, et al. Worldwide prevalence of hepatitis B virus and hepatitis C virus among patients with cirrhosis at country, region, and global levels: a systematic review. Lancet Gastroenterol Hepatol 2022; 7: 724-735.
20. Ho JK, Jeevan-Raj B, Netter HJ. Hepatitis B virus (HBV) subviral particles as protective vaccines and vaccine platforms. Viruses 2020; 12: 126.
21. Anutebeh EN, Tatah L, Feteh VF, Aroke D, Assob JCN, Choukem SP. Immune response to hepatitis B vaccine following complete immunization of children attending two regional hospitals in the Southwest region of Cameroon: a cross sectional study. BMC Infect Dis 2021; 21: 1205.
22. Hill HA, Elam-Evans LD, Yankey D, Singleton JA, Kolasa M. National, state, and selected local area vaccination coverage among children aged 19-35 months-United States, 2014. MMWR Morb Mortal Wkly Rep 2015; 64: 889-896.
23. Larijani MS, Sadat SM, Bolhassani A, Pouriayevali MH, Bahramali G, Ramezani A. In silico design and immunologic evaluation of HIV-1 p24-Nef fusion protein to approach a therapeutic vaccine candidate. Curr HIV Res 2018; 16: 322-337.
24. Sadat Larijani M, Ramezani A, Mashhadi Abolghasem Shirazi M, Bolhassani A, Pouriayevali MH, Shahbazi S, et al. Evaluation of transduced dendritic cells expressing HIV-1 p24-Nef antigens in HIV-specific cytotoxic T cells induction as a therapeutic candidate vaccine. Virus Res 2021; 298: 198403.
25. Beran J. Bivalent inactivated hepatitis A and recombinant hepatitis B vaccine. Expert Rev Vaccines 2007; 6: 891-902.
26. Senger T, Schädlich L, Gissmann L, Müller M. Enhanced papillomavirus-like particle production in insect cells. Virology 2009; 388: 344-353.
27. Nain A, Kumar M, Banerjee M. Oligomers of hepatitis A virus (HAV) capsid protein VP1 generated in a heterologous expression system. Microb Cell Fact 2022; 21: 53.
28. Feng R, Wang L, Shi D, Zheng B, Zhang L, Hou H. Structural basis for neutralization of an anicteric hepatitis associated echovirus by a potent neutralizing antibody. Cell Discov 2021; 7: 35.
29. Ahmadi N, Aghasadeghi M, Hamidi-Fard M, Motevalli F, Bahramali G. Reverse vaccinology and immunoinformatic approach for designing a bivalent vaccine candidate against hepatitis A and hepatitis B viruses. Mol Biotechnol 2024; 66: 2362-2380.
30. Ayyagari VS, T C V, K AP, Srirama K. Design of a multi-epitope-based vaccine targeting M-protein of SARS-CoV2: an immunoinformatics approach. J Biomol Struct Dyn 2022; 40: 2963-2977.
31. Chao P, Zhang X, Zhang L, Yang A, Wang Y, Chen X. Proteomics-based vaccine targets annotation and design of multi-epitope vaccine against antibiotic-resistant Streptococcus gallolyticus. Sci Rep 2024; 14: 4836.
32. Stuurman AL, Marano C, Bunge EM, De Moerlooze L, Shouval D. Impact of universal mass vaccination with monovalent inactivated hepatitis A vaccines–A systematic review. Hum Vaccin Immunother 2017; 13: 724-736.
33. Lemon SM, Ott JJ, Van Damme P, Shouval D. Type A viral hepatitis: A summary and update on the molecular virology, epidemiology, pathogenesis and prevention. J Hepatol 2017; S0168-8278(17)32278-X.
34. Nain A, Kumar M, Banerjee M. Oligomers of hepatitis A virus (HAV) capsid protein VP1 generated in a heterologous expression system. Microb Cell Fact 2022; 21: 53.
35. Larijani MS, Pouriayevali MH, Sadat SM, Ramezani A. Production of recombinant HIV-1 p24-Nef protein in two forms as potential candidate vaccines in three vehicles. Curr Drug Deliv 2020; 17: 387-395.
36. Ai J, Zhang H, Zhang Q, Zhang Y, Lin K, Fu Z, et al. Recombinant protein subunit vaccine booster following two-dose inactivated vaccines dramatically enhanced anti-RBD responses and neutralizing titers against SARS-CoV-2 and Variants of Concern. Cell Res 2022; 32: 103-106.
37. Beaumont E, Patient R, Hourioux C, Dimier-Poisson I, Roingeard P. Chimeric hepatitis B virus/hepatitis C virus envelope proteins elicit broadly neutralizing antibodies and constitute a potential bivalent prophylactic vaccine. Hepatology 2013; 57: 1303-1313.
38. Van Damme P, Van Herck K. A review of the efficacy, immunogenicity and tolerability of a combined hepatitis A and B vaccine. Expert Rev Vaccines 2004; 3: 249-267.
39. Jang KO, Park JH, Lee HH, Chung DK, Kim W, Chung IS. Expression and immunogenic analysis of recombinant polypeptides derived from capsid protein VP1 for developing subunit vaccine material against hepatitis A virus. Protein Expr Purif 2014; 100: 1-9.
40. Banerjee K, Klasse PJ, Sanders RW, Pereyra F, Michael E, Lu M, et al. IgG subclass profiles in infected HIV type 1 controllers and chronic progressors and in uninfected recipients of Env vaccines. AIDS Res Hum Retroviruses 2010; 26: 445-458.
41. Lee JM, Lee HH, Hwang-Bo J, Shon DH, Kim W, Chung IS. Expression and immunogenicity of recombinant polypeptide VP1 of human hepatitis A virus in stably transformed fruitfly (Drosophila melanogaster) Schneider 2 cells. Biotechnol Appl Biochem 2009; 53: 101-109.
42. Kang S, Brown HM, Hwang S. Direct antiviral mechanisms of interferon-gamma. Immune Netw 2018; 18(5): e33.
43. Shin EC, Sung PS, Park SH. Immune responses and immunopathology in acute and chronic viral hepatitis. Nat Rev Immunol 2016; 16: 509-523.
44. Chung HY, Lee HH, Kim KI, Chung HY, Hwang-Bo J, Park JH, et al. Expression of a recombinant chimeric protein of hepatitis A virus VP1-Fc using a replicating vector based on Beet curly top virus in tobacco leaves and its immunogenicity in mice. Plant Cell Rep 2011; 30: 1513-1521.
45. Mutiso JM, Macharia JC, Kariuki TM, Gicheru MM. Montanide ISA 720 is more effective than BCG as an adjuvant for Leishmania killed vaccine in BALB/c mice. Int J Integ Biol 2009; 7: 107-116.
46. Marques RF, de Melo FM, Novais JT, Soares IS, Bargieri DY, Gimenez AM. Immune system modulation by the adjuvants poly (I: C) and montanide ISA 720. Front Immunol 2022; 13: 910022.
47. Aucouturier J, Dupuis L, Deville S, Ascarateil S, Ganne V. Montanide ISA 720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines. Expert Rev Vaccines 2002; 1: 111-118.
48. Marques RF, de Melo FM, Novais JT, Soares IS, Bargieri DY, Gimenez AM. Immune system modulation by the adjuvants poly (I: C) and montanide ISA 720. Front Immunol 2022; 13: 910022.
| Files | ||
| Issue | Vol 17 No 4 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i4.19257 | |
| Keywords | ||
| Subunit vaccine Hepatitis A Hepatitis B Bivalent vaccine Hepatitis A virus VP1 protein Adjuvant | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Hannan M, Jabalameli L, Aghasadeghi MR, Harzandi N, Sadat SM. Immunogenicity assessment of Hepatitis A-VP1 and Hepatitis B surface antigen (HBsAg) fusion protein: a novel bivalent vaccine candidate. Iran J Microbiol. 2025;17(4):636-643.
