Evaluation of Heterologous Prime-boost Vaccine Strategy Using Full-length Cytomegalovirus Glycoprotein B to Trigger BALB/c Mice Immunity
Abstract
Human cytomegalovirus glycoprotein B (gB) emerges as a viable candidate for eliciting neutralizing antibodies. This research specifically focused on exploring the immune reaction prompted by the nonglycosylated variant of the gB, with a comprehensive assessment of humoral immunity in mice.
The gB coding sequence was optimized and expressed in pET-15b. Additionally, pcDNA3.1(+) vectors were also used for cloning the same gB sequence as the DNA vaccine. The gB was purified using a Ni-NTA chromatographic column. SDS-PAGE and Western blotting were used to confirm protein expression and purification. Using the prime-boost strategy, 8 different BALB/c mice were injected with DNA vaccine plus gB heterologous vaccine at 3 intervals. We evaluated the interferon (IFN-γ), interleukin (IL-4), immunoglobulin (Ig) G1, IgG2a, and IgG2b using enzyme-linked immunosorbent assay.
It was shown that the mice administered with DNA vaccine plus gB had higher IFN- γ and IL-4 levels compared to controls. On the other hand, the mice that received 3 doses of gB showed the highest levels of IgG1 and IgG2a. However, IgG2b was at its highest in mice administrated with DNA vaccine plus gB. The total IgG was higher in mice that received gB than in other interventions.
According to the findings, the DNA vaccine enhanced total IgG in immunized mice more effectively than the gB. This could be attributed to conformational changes owing to a lack of glycan moiety. Furthermore, combining nonglycosylated gB with DNA as a heterologous vaccine strategy enhances innate immunity by increasing the IFN- γ levels.
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15. Schrader J, letters GMI, 2007 undefined. Location, location, timing: analysis of cytomegalovirus epitopes for neutralizing antibodies. Elsevier.
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21. Kadkhodayan S, Jafarzade BS, Sadat SM, Motevalli F, Agi E, Bolhassani A. Combination of cell penetrating peptides and heterologous DNA prime/protein boost strategy enhances immune responses against HIV-1 Nef antigen in BALB/c mouse model. Immunology letters. 2017 Aug 1;188:38-45.
22. Gil A, Shen S, Coley S, Gibson L, Diamond DJ, Wang S, et al. DNA vaccine prime followed by boost with live attenuated virus significantly improves antigenspecific T cell responses against human cytomegalovirus. Human Hum Vaccin Immunother. 2013;9(10):2120–32.
23. Dioverti MV, Razonable RR. Cytomegalovirus. Diagnostic Microbiology of the Immunocompromised Host. 2016 Aug 15:97-125.
24. Gomes AC, Baraniak IA, Lankina A, Moulder Z, Holenya P, Atkinson C, et al. The cytomegalovirus gB/MF59 vaccine candidate induces antibodies against an antigenic domain controlling cell-to-cell spread. Nat Commun. 2023;14(1):1–12.
25. Shibamura M, Yoshikawa T, Yamada S, Inagaki T, Nguyen PHA, Fujii H, et al. Association of human cytomegalovirus (HCMV) neutralizing antibodies with antibodies to the HCMV glycoprotein complexes. Virol J. 2022;17(1):1–10.
26. Kropff B, Burkhardt C, Schott J, Nentwich J, Fisch T, Britt W, et al. Glycoprotein N of Human Cytomegalovirus Protects the Virus from Neutralizing Antibodies. PLoS Pathogens. 2012;8(10):e1002999.
27. Yamaguchi K, Shimizu H, Takahashi K, Nagatomo T, Nishimura T, Matsumoto M, Koshizuka T, Mori H, Inoue N, Torikai M. Characterization of epitopes of human monoclonal antibodies against cytomegalovirus glycoprotein B for neutralization and antibody-dependent phagocytosis. Vaccine. 2023 Jun 14.
28. Dasari V, Beckett K, Horsefield S, Ambalathingal G, Khanna R. A bivalent CMV vaccine formulated with human compatible TLR9 agonist CpG1018 elicits potent cellular and humoral immunity in HLA expressing mice. PLoS Pathogens. 2022;18(6):e1010403.
29. McVoy M, Lee R, Saccoccio F, Hartikka J, Vaccine LS, 2015 undefined. A cytomegalovirus DNA vaccine induces antibodies that block viral entry into fibroblasts and epithelial cells. Elsevier.
30. Pardieck IN, van Duikeren S, Veerkamp DMB, Brasem DJ, Redeker A, van Bergen J, et al. Dominant Antiviral CD8+ T Cell Responses Empower Prophylactic Antibody-Eliciting Vaccines Against Cytomegalovirus. Front Immunol. 2022 Jan 27;13:125.
2. Mirarab A, Mohebbi A, Javid N, Moradi A, Vakili MA, Tabarraei A. Human cytomegalovirus pUL97 drug-resistance mutations in congenitally neonates and HIV-infected, no-drug-treated patients. Future Virol. 2017;12(1):13–8.
3. Mirarab A, Mohebbi A, Moradi A, Javid N, Vakili MA, Tabarraei A. Frequent pUL27 Variations in HIV-Infected Patients. Intervirology. 2017;59(5–6):262–6.
4. Crawford L, Tempel R, Streblow D, reports CKS, 2017 undefined. Human cytomegalovirus induces cellular and humoral virus-specific immune responses in humanized BLT mice. nature.com.
5. Liu Y, Freed DC, Li L, Tang A, Li F, Murray EM, et al. A Replication-Defective Human Cytomegalovirus Vaccine Elicits Humoral Immune Responses Analogous to Those with Natural Infection. J Virol. 2019;93(23).
6. Murthy S, Hayward GS, Wheelan S, Forman MS, Ahn JH, Pass RF, et al. Detection of a single identical cytomegalovirus (CMV) strain in recently seroconverted young women. PLoS ONE. 2011;6(1).
7. Ross S, Arora N, Novak Z. Cytomegalovirus reinfections in healthy seroimmune women. J Infect Dis. 2010 Feb 1;201(3):386-9.
8. Selinsky C, Luke C, Wloch M, Geall A, Hermanson G, Kaslow D, et al. A DNA-based vaccine for the prevention of human cytomegalovirus-associated diseases. Hum Vaccin. 2005;1(1):16-23.
9. Baraniak I, Kropff B, G McLean, S Pichon, F Piras-Douce, et L. Epitope-specific humoral responses to human cytomegalovirus glycoprotein-B vaccine with MF59: anti-AD2 levels correlate with protection from viremia. academic.oup.com. J Infect Dis. 2018;217(12):1907-17.
10. Mach M, Kropff B, Dal Monte P, Britt W. Complex Formation by Human Cytomegalovirus Glycoproteins M (gpUL100) and N (gpUL73). J Virol. 2000;74(24):11881–92.
11. Scarpini S, Morigi F, Betti L, Dondi A, Biagi C, Lanari M. Development of a Vaccine against Human Cytomegalovirus: Advances, Barriers, and Implications for the Clinical Practice. Vaccines (Basel). 2021;9(6):551.
12. Cui X, Cao Z, Wang S, Lee R, Wang X, Vaccine HM, et al. Novel trimeric human cytomegalovirus glycoprotein B elicits a high-titer neutralizing antibody response. Elsevier.
13. Li F, Freed D, Tang A, Rustandi R, vaccines MTN, 2017 undefined. Complement enhances in vitro neutralizing potency of antibodies to human cytomegalovirus glycoprotein B (gB) and immune sera induced by gB/MF59. NPJ Vaccines. 2017:2:36.
14. Cayatte C, Schneider-Ohrum K, Wang Z, Irrinki A, Nguyen N, Lu J, et al. Cytomegalovirus Vaccine Strain Towne-Derived Dense Bodies Induce Broad Cellular Immune Responses and Neutralizing Antibodies That Prevent Infection of Fibroblasts and Epithelial Cells. J of Virol. 2013;87(20):11107–20.
15. Schrader J, letters GMI, 2007 undefined. Location, location, timing: analysis of cytomegalovirus epitopes for neutralizing antibodies. Elsevier.
16. Cui X, Cao Z, Wang S, Adler SP, McVoy MA, Snapper CM. Immunization with Human Cytomegalovirus Core Fusion Machinery and Accessory Envelope Proteins Elicit Strong Synergistic Neutralizing Activities. Vaccines. 2020;13;8(2):179.
17. Schleiss MR. Recombinant cytomegalovirus glycoprotein B vaccine: Rethinking the immunological basis of protection. Proc Natl Acad Sci U S A. 2018;115(24):6110–2.
18. Nelson CS, Herold BC, Permar SR. A new era in cytomegalovirus vaccinology: considerations for rational design of next-generation vaccines to prevent congenital cytomegalovirus infection. NPJ Vaccines. 2018;3(1):1–9.
19. Burke HG, Heldwein EE. Correction: Crystal Structure of the Human Cytomegalovirus Glycoprotein B. PLoS Pathogens. 2015;11(11):1005227.
20. Kadkhodayan S, Jafarzade B, Sadat S, letters FMI, 2017 undefined. Combination of cell penetrating peptides and heterologous DNA prime/protein boost strategy enhances immune responses against HIV-1 Nef antigen in BALB/c. Elsevier.
21. Kadkhodayan S, Jafarzade BS, Sadat SM, Motevalli F, Agi E, Bolhassani A. Combination of cell penetrating peptides and heterologous DNA prime/protein boost strategy enhances immune responses against HIV-1 Nef antigen in BALB/c mouse model. Immunology letters. 2017 Aug 1;188:38-45.
22. Gil A, Shen S, Coley S, Gibson L, Diamond DJ, Wang S, et al. DNA vaccine prime followed by boost with live attenuated virus significantly improves antigenspecific T cell responses against human cytomegalovirus. Human Hum Vaccin Immunother. 2013;9(10):2120–32.
23. Dioverti MV, Razonable RR. Cytomegalovirus. Diagnostic Microbiology of the Immunocompromised Host. 2016 Aug 15:97-125.
24. Gomes AC, Baraniak IA, Lankina A, Moulder Z, Holenya P, Atkinson C, et al. The cytomegalovirus gB/MF59 vaccine candidate induces antibodies against an antigenic domain controlling cell-to-cell spread. Nat Commun. 2023;14(1):1–12.
25. Shibamura M, Yoshikawa T, Yamada S, Inagaki T, Nguyen PHA, Fujii H, et al. Association of human cytomegalovirus (HCMV) neutralizing antibodies with antibodies to the HCMV glycoprotein complexes. Virol J. 2022;17(1):1–10.
26. Kropff B, Burkhardt C, Schott J, Nentwich J, Fisch T, Britt W, et al. Glycoprotein N of Human Cytomegalovirus Protects the Virus from Neutralizing Antibodies. PLoS Pathogens. 2012;8(10):e1002999.
27. Yamaguchi K, Shimizu H, Takahashi K, Nagatomo T, Nishimura T, Matsumoto M, Koshizuka T, Mori H, Inoue N, Torikai M. Characterization of epitopes of human monoclonal antibodies against cytomegalovirus glycoprotein B for neutralization and antibody-dependent phagocytosis. Vaccine. 2023 Jun 14.
28. Dasari V, Beckett K, Horsefield S, Ambalathingal G, Khanna R. A bivalent CMV vaccine formulated with human compatible TLR9 agonist CpG1018 elicits potent cellular and humoral immunity in HLA expressing mice. PLoS Pathogens. 2022;18(6):e1010403.
29. McVoy M, Lee R, Saccoccio F, Hartikka J, Vaccine LS, 2015 undefined. A cytomegalovirus DNA vaccine induces antibodies that block viral entry into fibroblasts and epithelial cells. Elsevier.
30. Pardieck IN, van Duikeren S, Veerkamp DMB, Brasem DJ, Redeker A, van Bergen J, et al. Dominant Antiviral CD8+ T Cell Responses Empower Prophylactic Antibody-Eliciting Vaccines Against Cytomegalovirus. Front Immunol. 2022 Jan 27;13:125.
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How to Cite
1.
Azadfar S, Yamchi A, Majd A, Tabarraei A. Evaluation of Heterologous Prime-boost Vaccine Strategy Using Full-length Cytomegalovirus Glycoprotein B to Trigger BALB/c Mice Immunity. Iran J Allergy Asthma Immunol. 2024;:1-10.
