PPD in HBsAg vaccine formulation suppressed IFN-γ and IL-4 cytokine responses and induced long-lived humoral immune responses: Results from 220-day monitoring of specific IgG responses

Document Type : Original Article

Authors

1 Department of Biotechnology, Faculty of Advanced Sciences & Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

2 Department of Microbiology, Faculty of Advanced Sciences & Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

3 Department of FMD vaccine, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

4 Department of Biology, Faculty of basic science, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

5 Advanced Therapy Medicinal Product (ATMP) Department, Breast Cancer Research Center, Motamed Cancer Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

6 Recombinant Vaccine Research Center, Tehran University of Medical Sciences, Tehran, Iran

7 Immunotherapy Group, The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences, Tehran, Iran

8 Department of Immunology, Pasteur Institute of Iran, Tehran, Iran

Abstract

Objective(s): HBsAg vaccine is unable to induce Th1 immune responses. Here, immune responses and long-lived IgG responses of HBsAg-Alum, HBsAg-MF59, as well as HBsAg-MF59 were compared when formulated with PPD. 
Materials and Methods: BALB/c mice were injected with the vaccines subcutaneously three times with a two-week interval. Then, specific IgG, long-lived IgG responses up to 220 days, and also IgG1 and IgG2a isotypes were assessed using ELISA. Furthermore, IFN-γ and IL-4 were assessed on spleen cell culture supernatant by ELISA. 
Results: IFN-γ cytokine response between MF59- and Alum-adjuvanted vaccines did not show a significant difference. HBsAg-Alum vaccine revealed an increase in IL-4 cytokine, as compared with those immunized with HBsAg-MF59 at borderline (P=0.0553). In addition, HBsAg-MF59+PPD 10 µg showed a significant decrease in IL-4 and IFN-γ cytokines, as compared with HBsAg-MF59. Furthermore, the HBsAg-MF59+PPD10 µg group showed a significant increase in the IL-2/IL-4 ratio, as compared with HBsAg-MF59 (P=0.0339). Specific IgG antibody showed a significant increase in HBsAg-MF59, as compared with HBsAg-Alum. Furthermore, HBsAg-MF59 plus PPD showed a significant increase in IgG responses, as compared with HBsAg-MF59 and HBsAg-Alum groups. Long-lived IgG responses up to 220 days after the final shot showed a significant increase in HBsAgMF59 versus HBsAg-Alum group and PPD in the HBsAg-MF59 vaccine formulation, resulting in a significant increase in IgG responses versus HBsAg-MF59 group. In addition, PPD in the HBsAg-MF59 vaccine formulation suppressed IgG1 response versus HBsAg-Alum. However, HBsAg-MF59 showed a significant increase in IgG2α versus the HBsAg-Alum group (P=0.0190). Immunization with HBsAg-MF59+PPD (10 µg) showed a significant increase versus the HBsAg-MF59 group (P=0.0040). Results from the IgG2a/IgG1 ratio in HBsAg-MF59+PPD1µg and HBsAg-MF59+PPD10 µg groups showed a significant increase as compared with HBsAg-MF59 groups (P<0.0345). 
Conclusion: PPD in the HBsAg vaccine formulation suppressed IL-4 and IFN-γ responses, but increased the IL-2/IL-4 ratio, IgG2a, and IgG2a/IgG1 responses, which may show Th1 polarization. Furthermore, PPD leads to more potent long-lived IgG responses in the HBsAg vaccine, highlighting its potential as a component of a complex adjuvant.

Keywords

References

1. Fakharzadeh S, Kalanaky S, Hafizi M, Goya MM, Masoumi Z, Namaki S, et al. The new nano-complex, Hep-c, improves the immunogenicity of the hepatitis B vaccine. Vaccine 2013; 31: 2591-2597. 
2. Mahdavi M, Mavandadnejad F, Yazdi MH, Faghfuri E, Hashemi H, Homayouni-Oreh S, et al. Oral administration of synthetic selenium nanoparticles induced robust Th1 cytokine pattern after HBs antigen vaccination in mouse model. J Infect Public Health 2017; 10: 102-109. 
3. Pollicino T, Amaddeo G, Restuccia A, Raffa G, Alibrandi A, Cutroneo G, et al. Impact of hepatitis B virus (HBV) preS/S genomic variability on HBV surface antigen and HBV DNA serum levels. Hepatology. 2012; 56: 434-443.
4. Hajarizadeh B, Mesgarpour B, Nasiri MJ, Alavian SM, Merat S, Poustchi H, et al. Estimating the prevalence of hepatitis B virus infection and exposure among general population in Iran. Hepat Mon 2017; 17: 11-21.
5. Mahdavi M, Ebtekar M, Khorshid HR, Azadmanesh K, Hartoonian C, Hassan ZM. ELISPOT analysis of a new CTL based DNA vaccine for HIV-1 using GM-CSF in DNA prime/peptide boost strategy: GM-CSF induced long-lived memory responses. Immunol Lett 2011; 140:14-20.
6. Amini Y, Tebianian M, Mosavari N, Fasihi Ramandi M, Ebrahimi SM, Najminejad H, et al. Development of an effective delivery system for intranasal immunization against Mycobacterium tuberculosis ESAT-6 antigen. Artif Cells Nanomed Biotechnol 2017; 45:291-296.
7. Najminejad H, Kalantar SM, Mokarram AR, Dabaghian M, Abdollahpour-Alitappeh M, Ebrahimi SM, et al. Bordetella pertussis antigens encapsulated into N-trimethyl chitosan nanoparticulate systems as a novel intranasal pertussis vaccine. Artif Cells Nanomed Biotechnol 2019; 47:2605-2611.
8. Jafarpour N, Memarnejadian A, Aghasadeghi MR, Kohram F, Aghababa H, Khoramabadi N, et al. Clustered epitopes within a new poly-epitopic HIV-1 DNA vaccine shows immunogenicity in BALB/c mice. Mol Biol Rep 2014; 41:5207-5214.
9. Xie Y, Sun HX, Li D. Platycodin D is a potent adjuvant of specific cellular and humoral immune responses against recombinant hepatitis B antigen. Vaccine. 2009; 27:757-764.
10. Roohvand F, Ehsani P, Abdollahpour-Alitappeh M, Shokri M, Kossari N. Biomedical applications of yeasts-a patent view, part two: era of humanized yeasts and expanded applications. Expert Opin Ther Pat 2020; 30:609-631.
11. 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; 34:516-522.
12. Jamali A, Mahdavi M, Shahabi S, Hassan ZM, Sabahi F, Javan M, et al. Naloxone, an opioid receptor antagonist, enhances induction of protective immunity against HSV-1 infection in BALB/c mice. Microb Pathog 2007; 43:217-223.
13. Lazarevic I, Banko A, Miljanovic D, Cupic M. Immune-escape hepatitis B virus mutations associated with viral reactivation upon immunosuppression. Viruses. 2019; 11:778-794.
14. Calabro S, Tritto E, Pezzotti A, Taccone M, Muzzi A, Bertholet S, et al. The adjuvant effect of MF59 is due to the oil-in-water emulsion formulation, none of the individual components induce a comparable adjuvant effect. Vaccine 2013; 31:3363-3369.
15. Bagheri K, Delirezh N, Moazzeni S-M. PPD extract induces the maturation of human monocyte-derived dendritic cells. Immunopharmacol Immunotoxicol 2008; 30:91-104.
16. Rezaei M, Hosseini SN, Khavari-Nejad RA, Najafi F, Mahdavi M. HBs antigen and mannose loading on the surface of iron oxide nanoparticles in order to immuno-targeting: fabrication, characterization, cellular and humoral immunoassay. Artif Cells Nanomed Biotechnol 2019; 47:1543-1558.
17. Abou‐Taleb DA, Abou‐Taleb HA, El‐Badawy O, Ahmed AO, Thabiet Hassan AE, Awad SM. Intralesional vitamin D3 versus intralesional purified protein derivative in treatment of multiple warts: a comparative clinical and immunological study. Dermatol Ther 2019; 32:e13034.
18. Kowalewicz-Kulbat M, Szpakowski P, Locht C, Biet F, Kaplonek P, Krawczyk KT, et al. Tuberculin skin test reaction is related to memory, but not naive CD4+ T cell responses to mycobacterial stimuli in BCG-vaccinated young adults. Vaccine 2018; 36:4566-4577.
19. Golkaran B, Mahdavi M. Application of PPD in the Formulation of Commercial Hepatitis B Vaccine to Improve Humoral and Cellular Immune Responses. Pathobiology Research. 2018; 21:73-78.
20. Reed SG, Coler RN, Dalemans W, Tan EV, DeLa Cruz EC, Basaraba RJ, et al. Defined tuberculosis vaccine, Mtb72F/AS02A, evidence of protection in cynomolgus monkeys. Proc Natl Acad Sci U S A 2009; 106:2301-2306.
21. Silveira H, Ordway D, Dockrell H, Jackson M, Ventura F. Cell‐mediated immune responses to mycobacterial antigens in patients with pulmonary tuberculosis and HIV infection. Clin Exp Immunol 1997; 110:26-34.
22. Mahdavi M, Ebtekar M, Azadmanesh K, Mahboudi F, Khorramkhorshid H, Rahbarizadeh F, et al. HIV-1 Gag p24-Nef fusion peptide induces cellular and humoral immune response in a mouse model. Acta virol 2010; 54:131-136.
23. Zadon S, Chaturvedi VK, Kumar B, Gupta PK, Mayank R, Varsha P. Effect on immune response against Pasteurella multocida capsular type A: 1 and A: 4 on supplementation with protein purified derivatives of Mycobacterium bovis in chicken. JEBAS 2017; 5:780-785.
24. Lewis D, Eiden J, Goilav C, Langenberg A, Suggett F, Griffin G. Rapid and frequent induction of protective immunity exceeding UK recommendations for healthcare settings by MF59-adjuvated hepatitis B vaccine. Commun Dis Public Health 2003; 6:320-324.
25. Heineman TC, Clements-Mann ML, Poland GA, Jacobson RM, Izu AE, Sakamoto D, et al. A randomized, controlled study in adults of the immunogenicity of a novel hepatitis B vaccine containing MF59 adjuvant. Vaccine. 1999; 17:2769-2778.
26. Traquina P, Morandi M, Contorni M, Van Nest G. MF59 adjuvant enhances the antibody response to recombinant hepatitis B surface antigen vaccine in primates. J Infect Dis 1996;174:1168-1175.
27. Gavillet BM, Eberhardt CS, Auderset F, Castellino F, Seubert A, Tregoning JS, et al. MF59 mediates its B cell adjuvanticity by promoting T follicular helper cells and thus germinal center responses in adult and early life. J Immunol 2015; 194:4836-4845. 
28. Riteau N, Radtke AJ, Shenderov K, Mittereder L, Oland SD, Hieny S, et al. Water-in-oil–only adjuvants selectively promote T follicular helper cell polarization through a type I IFN and IL-6–dependent pathway. J Immunol. 2016; 197:3884-3893. 
29. Wu C, Li Z, Fu X, Yu S, Lao S, Yang B. Antigen-specific human NKT cells from tuberculosis patients produce IL-21 to help B cells for the production of immunoglobulins. Oncotarget. 2015; 6:28633-28645. 
30. He J, Fan Y, Shen D, Yu M, Shi L, Ding S, Li L. Characterization of cytokine profile to distinguish latent tuberculosis from active tuberculosis and healthy controls. Cytokine 2020; 135:155218.
Iranian Journal of Basic Medical Sciences
Volume 25, Issue 11
November 2022
Pages 1326-1333

Files

Share

How to cite

Statistics