The heterologous expression of novel recombinant protein composed of HN and F moieties of Newcastle disease virus and immunogenicity evaluation in mouse model
-
Atena Mozafari
,
Mehregan Rahmani
,
Yasaman Yasini Nasab
,
Shahla Shahsavandi
,
Mahyat Jafari
,
Ali Hatef Salmanian
Abstract
Background and Objectives: The rapid spread of Newcastle disease (ND), driven by extensive commercial exchange in the poultry industry, necessitates urgent preventive measures. Although effective vaccines against the Newcastle disease virus (NDV) have been used since 1940, recent outbreaks and the limitations of current vaccines highlight the need for improved solutions. Advances in synthetic biology, reverse vaccinology, molecular biology, and recombinant DNA technology over the past 20 years have led to the development of recombinant vaccines, which offer enhanced protection and broader immunogenic coverage against NDV. This study aimed to express the immunogenic domains of Hemagglutinin Neuraminidase (HN) and Fusion (F) glycoproteins, linked to the heat-labile enterotoxin B subunit (LTB) bio-adjuvant, to develop an effective and reliable recombinant vaccine for NDV.
Materials and Methods: In this study, the L(HN)2F protein, composed of the LTB bio-adjuvant and the immunogenic regions of the doubled Hemagglutinin Neuraminidase (HN-HN) and Fusion (F) epitope, was expressed in Escherichia coli. Subcutaneous injection was used to evaluate the humoral immune response in mice and the result was compared with B1 vaccine.
Results: The induction of strong humoral immune responses proved the strong immunoreactivity of the recombinant protein.
Conclusion: The IgG elicited by the recombinant proteins was comparable to that of the commercial B1 vaccine against NDV, indicating its potential as a viable candidate for further development and evaluation as a recombinant vaccine against NDV.
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16. Da Hora VP, Conceição FR, Dellagostin OA, Doolan DL. Non-toxic derivatives of LT as potent adjuvants. Vaccine 2011; 29: 1538-1544.
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18. Peng X, Zhang R, Wang C, Yu F, Yu M, Chen S, et al. E. coli enterotoxin LtB enhances vaccine-induced Anti-H. pylori protection by promoting leukocyte migration into gastric mucus via inflammatory lesions. Cells 2019; 8: 982.
19. Lei H, Peng X, Shu H, Zhao D. Intranasal immunization with live recombinant Lactococcus lactis combined with heat-labile toxin B subunit protects chickens from highly pathogenic avian influenza H5N1 virus. J Med Virol 2015; 87: 39-44.
20. El-Shall NA, Shewita RS, Abd El-Hack ME, AlKahtane A, Alarifi S, Alkahtani S, et al. Effect of essential oils on the immune response to some viral vaccines in broiler chickens, with special reference to Newcastle disease virus. Poult Sci 2020; 99: 2944-2954.
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22. Gaikwad SS, Lee HJ, Kim JY, Choi KS. Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein. Clin Exp Vaccine Res 2019; 8: 27-34.
23. Mozafari A, Amani J, Shahsavandi S, Salmanian AH. A novel multi-epitope edible vaccine Candidate for Newcastle disease virus: In silico approach. Iran J Biotechnol 2022; 20(2): e3119.
24. Cheng Y, Wei H, Sun R, Tian Z, Zheng X. Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80. Anal Biochem 2016; 494: 37-39.
25. Park AR, Jang SW, Kim JS, Park YG, Koo BS, Lee HC. Efficient recovery of recombinant CRM197 expressed as inclusion bodies in E. coli. PLoS One 2018; 13(7): e0201060.
26. de Haro-Cruz MJ, Guadarrama-Macedo SI, López-Hurtado M, Escobedo-Guerra MR, Guerra-Infante FM. Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis. Int Microbiol 2019; 22: 471-478.
27. Shahid N, Rao A, Ahad A, Gul A, Latif A, Azam S, et al. E. coli expression and immunological assessment of expressed recombinant Newcastle disease virus hemagglutinin-neuraminidase protein in chickens. Acta Virol 2020; 64: 331-337.
28. Pollet J, Chen W-H, Strych U. Recombinant protein vaccines, a proven approach against coronavirus pandemics. Adv Drug Deliv Rev 2021; 170: 71-82.
29. Throngnumchai B, Jitrakorn S, Sangsuriya P, Unajak S, Khunrae P, Dong HT, et al. Refolded recombinant major capsid protein (MCP) from Infectious Spleen and Kidney Necrosis Virus (ISKNV) effectively stimulates serum specific antibody and immune related genes response in Nile tilapia (Oreochromis niloticus). Protein Expr Purif 2021; 184: 105876.
30. Mecha MF, Hutchinson RB, Lee JH, Cavagnero S. Protein folding in vitro and in the cell: From a solitary journey to a team effort. Biophys Chem 2022; 287: 106821.
31. Zykova AA, Blokhina EA, Kotlyarov RY, Stepanova LA, Tsybalova LM, Kuprianov VV, et al. Highly immunogenic nanoparticles based on a fusion protein comprising the M2e of influenza A virus and a Lipopeptide. Viruses 2020; 12: 1133.
32. Amani J, Salmanian AH, Rafati S, Mousavi SL. Immunogenic properties of chimeric protein from espA, eae and tir genes of Escherichia coli O157:H7. Vaccine 2010; 28: 6923-6929.
33. Motamedi MJ, Shahsavandi S, Amani J, Kazemi R, Takrim S, Jafari M, et al. Immunogenicity of the Multi-Epitopic recombinant glycoproteins of newcastle disease Virus: Implications for the serodiagnosis applications. Iran J Biotechnol 2018; 16(4): e1749.
2. Alsahami AA, Ideris A, Omar A, Ramanoon SZ, Sadiq MB. Isolation, identification and molecular characterization of Newcastle disease viruses in vaccinated chickens from commercial farms in the Sultanate of Oman. Int J Vet Sci Med 2018; 6: 248-252.
3. Ma F, Zhang E, Li Q, Xu Q, Ou J, Yin H, et al. A Plant-produced recombinant fusion protein-based Newcastle disease subunit vaccine and rapid differential diagnosis platform. Vaccines (Basel) 2020; 8: 122.
4. Bello MB, Yusoff K, Ideris A, Hair-Bejo M, Jibril AH, Peeters BP, et al. Exploring the prospects of engineered Newcastle disease virus in modern vaccinology. Viruses 2020; 12: 451.
5. Bello MB, Mahamud SNA, Yusoff K, Ideris A, Hair-Bejo M, Peeters BPH, et al. Development of an effective and stable genotype-matched live attenuated Newcastle disease virus vaccine based on a novel naturally recombinant Malaysian isolate using reverse genetics. Vaccines (Basel) 2020; 8: 270.
6. Dimitrov KM, Afonso CL, Yu Q, Miller PJ. Newcastle disease vaccines—A solved problem or a continuous challenge? Vet Microbiol 2017; 206: 126-136.
7. Jawale CV, Lee JH. Characterization of a Salmonella Typhimurium ghost carrying an adjuvant protein as a vaccine candidate for the protection of chickens against virulent challenge. Avian Pathol 2014; 43: 506-513.
8. Viktorova EG, Khattar SK, Kouiavskaia D, Laassri M, Zagorodnyaya T, Dragunsky E, et al. Newcastle disease virus-based vectored vaccine against poliomyelitis. J Virol 2018; 92(17): e00976-18.
9. Nurzijah I, Elbohy OA, Kanyuka K, Daly JM, Dunham S. Development of plant-based vaccines for prevention of avian influenza and Newcastle disease in poultry. Vaccines (Basel) 2022; 10: 478.
10. Kang X, Wang J, Jiao Y, Tang P, Song L, Xiong D, et al. Expression of recombinant Newcastle disease virus F protein in Pichia pastoris and its immunogenicity using flagellin as the adjuvant. Protein Expr Purif 2016; 128: 73-80.
11. Shahid N, Samiullah TR, Shakoor S, Latif A, Yasmeen A, Azam S, et al. Early stage development of a Newcastle disease vaccine candidate in corn. Front Vet Sci 2020; 7: 499.
12. Mayahi V, Esmaelizad M, Harzandi N. Designing a novel recombinant HN protein with multi neutralizing antigenic sites and auto tag removal ability based on NDV-VIIj for diagnosis and vaccination application. Indian J Microbiol 2018; 58: 326-331.
13. Boroujeni NA, Khatouni SB, Motamedi MJ, Afraz S, Jafari M, Salmanian A-H. Root-preferential expression of Newcastle virus glycoproteins driven by NtREL1 promoter in tobacco hairy roots and evaluation of oral delivery in mice. Transgenic Res 2022; 31: 201-213.
14. Motamedi MJ, Ebrahimi MM, Shahsavandi S, Amani J, Kazemi R, Jafari M, et al. The immunogenicity of a novel chimeric Hemagglutinin-neuraminidase-fusion antigen from Newcastle disease virus by oral delivery of transgenic canola seeds to chickens. Mol Biotechnol 2020; 62: 344-354.
15. Draper SJ, Angov E, Horii T, Miller LH, Srinivasan P, Theisen M, et al. Recent advances in recombinant protein-based malaria vaccines. Vaccine 2015; 33: 7433-7443.
16. Da Hora VP, Conceição FR, Dellagostin OA, Doolan DL. Non-toxic derivatives of LT as potent adjuvants. Vaccine 2011; 29: 1538-1544.
17. Zerna G, Rathinasamy VA, Toet H, Anderson G, Dempster R, Spithill TW, et al. Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle. Vaccines (Basel) 2021; 9: 1213.
18. Peng X, Zhang R, Wang C, Yu F, Yu M, Chen S, et al. E. coli enterotoxin LtB enhances vaccine-induced Anti-H. pylori protection by promoting leukocyte migration into gastric mucus via inflammatory lesions. Cells 2019; 8: 982.
19. Lei H, Peng X, Shu H, Zhao D. Intranasal immunization with live recombinant Lactococcus lactis combined with heat-labile toxin B subunit protects chickens from highly pathogenic avian influenza H5N1 virus. J Med Virol 2015; 87: 39-44.
20. El-Shall NA, Shewita RS, Abd El-Hack ME, AlKahtane A, Alarifi S, Alkahtani S, et al. Effect of essential oils on the immune response to some viral vaccines in broiler chickens, with special reference to Newcastle disease virus. Poult Sci 2020; 99: 2944-2954.
21. Blokhina EA, Mardanova ES, Stepanova LA, Tsybalova LM, Ravin NV. Plant-produced recombinant influenza a virus Candidate vaccine based on Flagellin linked to conservative fragments of M2 protein and Hemagglutintin. Plants (Basel) 2020; 9: 162.
22. Gaikwad SS, Lee HJ, Kim JY, Choi KS. Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein. Clin Exp Vaccine Res 2019; 8: 27-34.
23. Mozafari A, Amani J, Shahsavandi S, Salmanian AH. A novel multi-epitope edible vaccine Candidate for Newcastle disease virus: In silico approach. Iran J Biotechnol 2022; 20(2): e3119.
24. Cheng Y, Wei H, Sun R, Tian Z, Zheng X. Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80. Anal Biochem 2016; 494: 37-39.
25. Park AR, Jang SW, Kim JS, Park YG, Koo BS, Lee HC. Efficient recovery of recombinant CRM197 expressed as inclusion bodies in E. coli. PLoS One 2018; 13(7): e0201060.
26. de Haro-Cruz MJ, Guadarrama-Macedo SI, López-Hurtado M, Escobedo-Guerra MR, Guerra-Infante FM. Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis. Int Microbiol 2019; 22: 471-478.
27. Shahid N, Rao A, Ahad A, Gul A, Latif A, Azam S, et al. E. coli expression and immunological assessment of expressed recombinant Newcastle disease virus hemagglutinin-neuraminidase protein in chickens. Acta Virol 2020; 64: 331-337.
28. Pollet J, Chen W-H, Strych U. Recombinant protein vaccines, a proven approach against coronavirus pandemics. Adv Drug Deliv Rev 2021; 170: 71-82.
29. Throngnumchai B, Jitrakorn S, Sangsuriya P, Unajak S, Khunrae P, Dong HT, et al. Refolded recombinant major capsid protein (MCP) from Infectious Spleen and Kidney Necrosis Virus (ISKNV) effectively stimulates serum specific antibody and immune related genes response in Nile tilapia (Oreochromis niloticus). Protein Expr Purif 2021; 184: 105876.
30. Mecha MF, Hutchinson RB, Lee JH, Cavagnero S. Protein folding in vitro and in the cell: From a solitary journey to a team effort. Biophys Chem 2022; 287: 106821.
31. Zykova AA, Blokhina EA, Kotlyarov RY, Stepanova LA, Tsybalova LM, Kuprianov VV, et al. Highly immunogenic nanoparticles based on a fusion protein comprising the M2e of influenza A virus and a Lipopeptide. Viruses 2020; 12: 1133.
32. Amani J, Salmanian AH, Rafati S, Mousavi SL. Immunogenic properties of chimeric protein from espA, eae and tir genes of Escherichia coli O157:H7. Vaccine 2010; 28: 6923-6929.
33. Motamedi MJ, Shahsavandi S, Amani J, Kazemi R, Takrim S, Jafari M, et al. Immunogenicity of the Multi-Epitopic recombinant glycoproteins of newcastle disease Virus: Implications for the serodiagnosis applications. Iran J Biotechnol 2018; 16(4): e1749.
| Files | ||
| Issue | Vol 16 No 5 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i5.16801 | |
| Keywords | ||
| Newcastle disease; Hemagglutinin neuraminidase; Fusion (F) glycoprotein; Immune response; LTB bio-adjuvant | ||
| Rights and permissions | |
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Mozafari A, Rahmani M, Yasini Nasab Y, Shahsavandi S, Jafari M, Salmanian AH. The heterologous expression of novel recombinant protein composed of HN and F moieties of Newcastle disease virus and immunogenicity evaluation in mouse model. Iran J Microbiol. 2024;16(5):655-665.
