Designing the fusion protein of rotavirus VP8 and hepatitis A virus VP1 and evaluating the immunological response in BALB/c mice
-
Hassan Yarmohammadi
,
Mohammadreza Aghasadeghi
,
Abbas Akhavan Sepahi
,
Mojtaba Hamidi-fard
,
Golnaz Bahramali
Abstract
Background and Objectives: Rotavirus and Hepatitis A virus are responsible for causing gastroenteritis and jaundice. The current vaccination approaches have proven insufficient, especially in low-income countries. In this study, we presented a novel dual-vaccine candidate that combines the rotavirus VP8 protein and the hepatitis A virus VP1.
Materials and Methods: The VP8*-rotavirus+AAY+HAV-VP1 fusion protein was produced using an Escherichia coli expression system. The recombinant protein had a molecular weight of approximately 45.5 kDa and was purified through affinity chromatography. BALB/c mice were injected subcutaneously with the recombinant protein, VP1, VP8 and vaccines for rotavirus and hepatitis A virus, both with and without ALUM and M720 adjuvants. ELISA assays were used to measure total IgG, IgG1, IgG2, and short-term and long-term IL-5 and IFN-γ responses.
Results: The fusion protein, when combined with adjuvants, elicited significantly higher total IgG, IgG1, and IgG2 responses compared to VP1 and VP8 alone, as well as the rotavirus and hepatitis A vaccines. Furthermore, it induced a higher short-term IL-5 and IFN-γ response while demonstrating a higher long-term IL-5 response compared to the rotavirus and hepatitis A vaccines.
Conclusion: This study demonstrates that the VP8*-rotavirus+AAY+HAV-VP1 fusion protein is a promising dual vaccine candidate for immunization against hepatitis A and rotaviruses.
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7. Yen C, Jakob K, Esona MD, Peckham X, Rausch J, Hull JJ, et al. Detection of fecal shedding of rotavirus vaccine in infants following their first dose of pentavalent rotavirus vaccine. Vaccine 2011; 29: 4151-4155.
8. Payne DC, Edwards KM, Bowen MD, Keckley E, Peters J, Esona MD, et al. Sibling transmission of vaccine-derived rotavirus (RotaTeq) associated with rotavirus gastroenteritis. Pediatrics 2010; 125(2): e438-e441.
9. Pesavento JB, Crawford SE, Estes MK, Prasad BV. Rotavirus proteins: structure and assembly. Curr Top Microbiol Immunol 2006; 309: 189-219.
10. López S, Arias CF. Multistep entry of rotavirus into cells: a Versaillesque dance. Trends Microbiol 2004; 12: 271-278.
11. Arias CF, Silva-Ayala D, López S. Rotavirus entry: a deep journey into the cell with several exits. J Virol 2015; 89: 890-893.
12. Wen X, Cao D, Jones RW, Li J, Szu S, Hoshino Y. Construction and characterization of human rotavirus recombinant VP8* subunit parenteral vaccine candidates. Vaccine 2012; 30: 6121-6126.
13. Wen X, Wen K, Cao D, Li G, Jones RW, Li J, et al. Inclusion of a universal tetanus toxoid CD4(+) T cell epitope P2 significantly enhanced the immunogenicity of recombinant rotavirus DeltaVP8* subunit parenteral vaccines. Vaccine 2014; 32: 4420-4427.
14. Burke RM, Tate JE, Kirkwood CD, Steele AD, Parashar UD. Current and new rotavirus vaccines. Curr Opin Infect Dis 2019; 32: 435-444.
15. Cates JE, Tate JE, Parashar U. Rotavirus vaccines: progress and new developments. Expert Opin Biol Ther 2022; 22: 423-432.
16. Dormitzer PR, Sun Z-Y, Wagner G, Harrison SC. The rhesus rotavirus VP4 sialic acid binding domain has a galectin fold with a novel carbohydrate binding site. EMBO J 2002; 21: 885-897.
17. Zeller M, Patton JT, Heylen E, De Coster S, Ciarlet M, Van Ranst M, et al. Genetic analyses reveal differences in the VP7 and VP4 antigenic epitopes between human rotaviruses circulating in Belgium and rotaviruses in Rotarix and RotaTeq. J Clin Microbiol 2012; 50: 966-976.
18. Gholizadeh O, Akbarzadeh S, Ghazanfari Hashemi M, Gholami M, Amini P, Yekanipour Z, et al. Hepatitis A: viral structure, classification, life cycle, clinical symptoms, diagnosis error, and vaccination. Can J Infect Dis Med Microbiol 2023; 2023: 4263309.
19. Dennehy PH. Transmission of rotavirus and other enteric pathogens in the home. Pediatr Infect Dis J 2000; 19(10 Suppl): S103-S105.
20. Matheny SC, Kingery JE. Hepatitis A. Am Fam Physician 2012; 86: 1027-1034; quiz 1010-1012.
21. Severi E, Georgalis L, Pijnacker R, Veneti L, Turiac IA, Chiesa F, et al. Severity of the clinical presentation of hepatitis A in five European countries from 1995 to 2014. Int J Infect Dis 2022; 118: 34-43.
22. McKnight KL, Lemon SM. Hepatitis A virus genome organization and replication strategy. Cold Spring Harb Perspect Med 2018; 8: a033480.
23. 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.
24. Su Q, Guo M, Jia Z, Qiu F, Lu X, Gao Y, et al. Epitope-based recombinant diagnostic antigen to distinguish natural infection from vaccination with hepatitis A virus vaccines. J Virol Methods 2016; 233: 41-45.
25. 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 2023; 10.1007/s12033-023-00867-z.
26. 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.
27. 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.
28. Silva HC Junior, Pestana CP, Galler R, Medeiros MA. Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system. Mem Inst Oswaldo Cruz 2016; 111: 535-538.
29. Latifi T, Jalilvand S, Golsaz-Shirazi F, Arashkia A, Kachooei A, Afchangi A, et al. Characterization and immunogenicity of a novel chimeric hepatitis B core-virus like particles (cVLPs) carrying rotavirus VP8*protein in mice model. Virology 2023; 588: 109903.
30. Contreras JF, Menchaca GE, Infante R, Hernández CE, Rodríguez C, Tamez RS. Evidence of conserved epitopes in variable region of VP8* subunit of VP4 protein of rotaviruses of P(8)-1 and P(8)-3 lineages. Acta Virol 2011; 55: 273-278.
31. Feng N, Hu L, Ding S, Sanyal M, Zhao B, Sankaran B, et al. Human VP8* mAbs neutralize rotavirus selectively in human intestinal epithelial cells. J Clin Invest 2019; 129: 3839-3851.
32. Nair N, Feng N, Blum L K, Sanyal M, Ding S, Jiang B, et al. VP4-and VP7-specific antibodies mediate heterotypic immunity to rotavirus in humans. Sci Transl Med 2017; 9: eaam5434.
33. Kovacs-Nolan J, Mine Y. Tandem copies of a human rotavirus VP8 epitope can induce specific neutralizing antibodies in BALB/c mice. Biochim Biophys Acta 2006; 1760: 1884-1893.
34. Xia M, Wei C, Wang L, Cao D, Meng X-J, Jiang X, et al. Development and evaluation of two subunit vaccine candidates containing antigens of hepatitis E virus, rotavirus, and astrovirus. Sci Rep 2016; 6: 25735.
35. Takatsu K. Interleukin-5 and IL-5 receptor in health and diseases. Proc Jpn Acad Ser B Phys Biol Sci 2011; 87: 463-485.
36. Jorgovanovic D, Song M, Wang L, Zhang Y. Roles of IFN-γ in tumor progression and regression: a review. Biomark Res 2020; 8: 49.
37. Primorac D, Brlek P, Pavelic ES, Mesic J, Glavas Weinberger D, Matisic V, et al. Importance of cellular immunity and IFN-gamma concentration in preventing SARS-CoV-2 infection and reinfection: a cohort study. Viruses 2023; 15: 792.
38. Huang Y-L, Lin T-M, Wang S-Y, Wang S-R. The role of conserved arginine and proline residues in enterovirus VP1 protein. J Microbiol Immunol Infect 2022; 55: 590-597.
39. Favacho AR, Kurtenbach E, Sardi SI, Gouvea VS. Cloning, expression, and purification of recombinant bovine rotavirus hemagglutinin, VP8*, in Escherichia coli. Protein Expr Purif 2006; 46: 196-203.
2. Thanawastien A, Cartee RT, Griffin TJ 4th, Killeen KP, Mekalanos JJ. Conjugate-like immunogens produced as protein capsular matrix vaccines. Proc Natl Acad Sci U S A 2015; 112(10): E1143-E1151.
3. Crawford SE, Ramani S, Tate JE, Parashar UD, Svensson L, Hagbom M, et al. Rotavirus infection. Nat Rev Dis Primers 2017; 3: 17083.
4. Dóró R, László B, Martella V, Leshem E, Gentsch J, Parashar U, et al. Review of global rotavirus strain prevalence data from six years post vaccine licensure surveillance: is there evidence of strain selection from vaccine pressure? Infect Genet Evol 2014; 28: 446-461.
5. Cunliffe NA, Witte D, Ngwira BM, Todd S, Bostock NJ, Turner AM, et al. Efficacy of human rotavirus vaccine against severe gastroenteritis in Malawian children in the first two years of life: a randomized, double-blind, placebo controlled trial. Vaccine 2012; 30 Suppl 1(0 1): A36-A43.
6. Zaman K, Dang DA, Victor JC, Shin S, Yunus M, Dallas MJ, et al. Efficacy of pentavalent rotavirus vaccine against severe rotavirus gastroenteritis in infants in developing countries in Asia: a randomised, double-blind, placebo-controlled trial. Lancet 2010; 376: 615-623.
7. Yen C, Jakob K, Esona MD, Peckham X, Rausch J, Hull JJ, et al. Detection of fecal shedding of rotavirus vaccine in infants following their first dose of pentavalent rotavirus vaccine. Vaccine 2011; 29: 4151-4155.
8. Payne DC, Edwards KM, Bowen MD, Keckley E, Peters J, Esona MD, et al. Sibling transmission of vaccine-derived rotavirus (RotaTeq) associated with rotavirus gastroenteritis. Pediatrics 2010; 125(2): e438-e441.
9. Pesavento JB, Crawford SE, Estes MK, Prasad BV. Rotavirus proteins: structure and assembly. Curr Top Microbiol Immunol 2006; 309: 189-219.
10. López S, Arias CF. Multistep entry of rotavirus into cells: a Versaillesque dance. Trends Microbiol 2004; 12: 271-278.
11. Arias CF, Silva-Ayala D, López S. Rotavirus entry: a deep journey into the cell with several exits. J Virol 2015; 89: 890-893.
12. Wen X, Cao D, Jones RW, Li J, Szu S, Hoshino Y. Construction and characterization of human rotavirus recombinant VP8* subunit parenteral vaccine candidates. Vaccine 2012; 30: 6121-6126.
13. Wen X, Wen K, Cao D, Li G, Jones RW, Li J, et al. Inclusion of a universal tetanus toxoid CD4(+) T cell epitope P2 significantly enhanced the immunogenicity of recombinant rotavirus DeltaVP8* subunit parenteral vaccines. Vaccine 2014; 32: 4420-4427.
14. Burke RM, Tate JE, Kirkwood CD, Steele AD, Parashar UD. Current and new rotavirus vaccines. Curr Opin Infect Dis 2019; 32: 435-444.
15. Cates JE, Tate JE, Parashar U. Rotavirus vaccines: progress and new developments. Expert Opin Biol Ther 2022; 22: 423-432.
16. Dormitzer PR, Sun Z-Y, Wagner G, Harrison SC. The rhesus rotavirus VP4 sialic acid binding domain has a galectin fold with a novel carbohydrate binding site. EMBO J 2002; 21: 885-897.
17. Zeller M, Patton JT, Heylen E, De Coster S, Ciarlet M, Van Ranst M, et al. Genetic analyses reveal differences in the VP7 and VP4 antigenic epitopes between human rotaviruses circulating in Belgium and rotaviruses in Rotarix and RotaTeq. J Clin Microbiol 2012; 50: 966-976.
18. Gholizadeh O, Akbarzadeh S, Ghazanfari Hashemi M, Gholami M, Amini P, Yekanipour Z, et al. Hepatitis A: viral structure, classification, life cycle, clinical symptoms, diagnosis error, and vaccination. Can J Infect Dis Med Microbiol 2023; 2023: 4263309.
19. Dennehy PH. Transmission of rotavirus and other enteric pathogens in the home. Pediatr Infect Dis J 2000; 19(10 Suppl): S103-S105.
20. Matheny SC, Kingery JE. Hepatitis A. Am Fam Physician 2012; 86: 1027-1034; quiz 1010-1012.
21. Severi E, Georgalis L, Pijnacker R, Veneti L, Turiac IA, Chiesa F, et al. Severity of the clinical presentation of hepatitis A in five European countries from 1995 to 2014. Int J Infect Dis 2022; 118: 34-43.
22. McKnight KL, Lemon SM. Hepatitis A virus genome organization and replication strategy. Cold Spring Harb Perspect Med 2018; 8: a033480.
23. 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.
24. Su Q, Guo M, Jia Z, Qiu F, Lu X, Gao Y, et al. Epitope-based recombinant diagnostic antigen to distinguish natural infection from vaccination with hepatitis A virus vaccines. J Virol Methods 2016; 233: 41-45.
25. 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 2023; 10.1007/s12033-023-00867-z.
26. 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.
27. 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.
28. Silva HC Junior, Pestana CP, Galler R, Medeiros MA. Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system. Mem Inst Oswaldo Cruz 2016; 111: 535-538.
29. Latifi T, Jalilvand S, Golsaz-Shirazi F, Arashkia A, Kachooei A, Afchangi A, et al. Characterization and immunogenicity of a novel chimeric hepatitis B core-virus like particles (cVLPs) carrying rotavirus VP8*protein in mice model. Virology 2023; 588: 109903.
30. Contreras JF, Menchaca GE, Infante R, Hernández CE, Rodríguez C, Tamez RS. Evidence of conserved epitopes in variable region of VP8* subunit of VP4 protein of rotaviruses of P(8)-1 and P(8)-3 lineages. Acta Virol 2011; 55: 273-278.
31. Feng N, Hu L, Ding S, Sanyal M, Zhao B, Sankaran B, et al. Human VP8* mAbs neutralize rotavirus selectively in human intestinal epithelial cells. J Clin Invest 2019; 129: 3839-3851.
32. Nair N, Feng N, Blum L K, Sanyal M, Ding S, Jiang B, et al. VP4-and VP7-specific antibodies mediate heterotypic immunity to rotavirus in humans. Sci Transl Med 2017; 9: eaam5434.
33. Kovacs-Nolan J, Mine Y. Tandem copies of a human rotavirus VP8 epitope can induce specific neutralizing antibodies in BALB/c mice. Biochim Biophys Acta 2006; 1760: 1884-1893.
34. Xia M, Wei C, Wang L, Cao D, Meng X-J, Jiang X, et al. Development and evaluation of two subunit vaccine candidates containing antigens of hepatitis E virus, rotavirus, and astrovirus. Sci Rep 2016; 6: 25735.
35. Takatsu K. Interleukin-5 and IL-5 receptor in health and diseases. Proc Jpn Acad Ser B Phys Biol Sci 2011; 87: 463-485.
36. Jorgovanovic D, Song M, Wang L, Zhang Y. Roles of IFN-γ in tumor progression and regression: a review. Biomark Res 2020; 8: 49.
37. Primorac D, Brlek P, Pavelic ES, Mesic J, Glavas Weinberger D, Matisic V, et al. Importance of cellular immunity and IFN-gamma concentration in preventing SARS-CoV-2 infection and reinfection: a cohort study. Viruses 2023; 15: 792.
38. Huang Y-L, Lin T-M, Wang S-Y, Wang S-R. The role of conserved arginine and proline residues in enterovirus VP1 protein. J Microbiol Immunol Infect 2022; 55: 590-597.
39. Favacho AR, Kurtenbach E, Sardi SI, Gouvea VS. Cloning, expression, and purification of recombinant bovine rotavirus hemagglutinin, VP8*, in Escherichia coli. Protein Expr Purif 2006; 46: 196-203.
| Files | ||
| Issue | Vol 16 No 3 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i3.15797 | |
| Keywords | ||
| Recombinant fusion proteins; Immunoinformatics; Rotavirus; Hepatitis A virus; Bivalent vaccines; BALB/c mouse | ||
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How to Cite
1.
Yarmohammadi H, Aghasadeghi M, Akhavan Sepahi A, Hamidi-fard M, Bahramali G. Designing the fusion protein of rotavirus VP8 and hepatitis A virus VP1 and evaluating the immunological response in BALB/c mice. Iran J Microbiol. 2024;16(3):401-410.
