Enhancement of the immunogenicity of a Mycobacterium tuberculosis fusion protein using ISCOMATRIX and PLUSCOM nano-adjuvants as prophylactic vaccine after nasal administration in mice

Document Type : Original Article


1 Department of Laboratory Sciences, School of Allied Medical Sciences, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

2 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Medical Bacteriology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

5 Immunobiochemistry laboratory, Immunology Research Center, Bu-Ali Research Institute, Mashhad, Iran

6 Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad Iran


Objective(s): Tuberculosis (TB), a contagious disease caused by Mycobacterium tuberculosis                          (M. tuberculosis), remains a health problem worldwide and this infection has the highest mortality rate among bacterial infections. Current studies suggest that intranasal administration of new TB vaccines could enhance the immunogenicity of M. tuberculosis antigens. Hence, we aim to evaluate the protective efficacy and immunogenicity of HspX/EsxS fusion protein of M. tuberculosis along with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA through intranasal administration in a mice model.
Materials and Methods: In the present study, the recombinant fusion protein was expressed in Escherichia coli and purified and used to prepare different nanoparticle formulations in combination with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA. Mice were intranasally vaccinated with each formulation three times at an interval of 2 weeks. Three weeks after the final vaccination, IFN-γ, IL-4. IL-17, and TGF-β concentrations in the supernatant of cultured splenocytes of vaccinated mice as well as serum titers of IgG1 and IgG2a and sIgA titers in nasal lavage were determined.
Results: According to obtained results, intranasally vaccinated mice with formulations containing ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA could effectively induce IFN-γ and sIgA responses. Moreover, both HspX/EsxS/ISCOMATRIX/MPLA and HspX/EsxS/PLUSCOM/MPLA and their BCG booster formulation could strongly stimulate the immune system and enhance the immunogenicity of M. tuberculosis antigens.
Conclusion: The results demonstrate the potential of HspX/EsxS-fused protein in combination with ISCOMATRIX, PLUSCOM, and MPLA after nasal administration in enhancing the immune response against M. tuberculosis antigens. Both nanoparticles were good adjuvants in order to promote the immunogenicity of TB-fused antigens. So, nasal immunization with these formulations, could induce immune responses and be considered a new TB vaccine or a BCG booster.


Main Subjects

1. Organization WH. Global status report on alcohol and health 2018: World Health Organization; 2019.
2. Avarvand AY, Khademi F, Tafaghodi M, Ahmadipour Z, Moradi B, Meshkat Z. The roles of latency-associated antigens in tuberculosis vaccines. Indian J Tuberc 2019;66:487-491.
3. Yousefi-Avarvand A, Tafaghodi M, Soleimanpour S, Khademi F. HspX protein as a candidate vaccine against Mycobacterium tuberculosis: An overview. Front Biol 2018;13:293-296.
4. Khademi F, Yousefi-Avarvand A, Derakhshan M, Vaez H, Sadeghi R. Middle east Mycobacterium tuberculosis antibiotic resistance: A systematic review and meta-analysis. Infect Epidemio Med 2017;3:25-35.
5. Ghafoor T, Ikram A, Abbassi SA, Mirza IA, Hussain A, ullah Khan I, et al. Antimicrobial sensitivity pattern of clinical isolates of Mycobacterium tuberculosis: A retrospective study from a reference laboratory in Pakistan. J Virol Microbiol 2014;2014:1-6.
6. Liang J, Teng X, Yuan X, Zhang Y, Shi C, Yue T, et al. Enhanced and durable protective immune responses induced by a cocktail of recombinant BCG strains expressing antigens of multistage of Mycobacterium tuberculosis. Mol Immunol 2015;66:392-401.
7. Andersen P, Kaufmann SH. Novel vaccination strategies against tuberculosis. Cold Spring Harb Perspect Med 2014;4:a018523.
8. Costa ACd, Costa-Junior AdO, Oliveira FMd, Nogueira SV, Rosa JD, Resende DP, et al. A new recombinant BCG vaccine induces specific Th17 and Th1 effector cells with higher protective efficacy against tuberculosis. PLoS One 2014;9:e112848.
9. Soleimanpour S, Farsiani H, Mosavat A, Ghazvini K, Eydgahi MRA, Sankian M, et al. APC targeting enhances immunogenicity of a novel multistage Fc-fusion tuberculosis vaccine in mice. Appl Microbiol Biotechnol  2015;99:10467-10480.
10. Khademi F, Derakhshan M, Yousefi-Avarvand A, Tafaghodi M, Soleimanpour S. Multi-stage subunit vaccines against Mycobacterium tuberculosis: An alternative to the BCG vaccine or a BCG-prime boost? Expert Review of Vaccines 2018;17:31-44.
11. Shirvani F, Karimi A, Rajabnejad M. BCG vaccination as a prevention strategy, threats and benefits. Arch Pediatr Infect Dis 2016;4:e30180.
12. Triccas JA, Counoupas C. Novel vaccination approaches to prevent tuberculosis in children. Pneumonia 2016;8:1-7.
13. McHugh KJ, Guarecuco R, Langer R, Jaklenec A. Single-injection vaccines: Progress, challenges, and opportunities. J Control Release 2015 Dec 10;219:596-609.
14. Khademi F, Taheri RA, Avarvand AY, Vaez H, Momtazi-Borojeni AA, Soleimanpour S. Are chitosan natural polymers suitable as adjuvant/delivery system for anti-tuberculosis vaccines? Microb Pathog 2018;121:218-223.
15. Khademi F, Derakhshan M, Yousefi-Avarvand A, Tafaghodi M. Potential of polymeric particles as future vaccine delivery systems/adjuvants for parenteral and non-parenteral immunization against tuberculosis: A systematic review. Iran J Basic Med Sci 2018;21:116-123
16. Khademi F, Yousefi-Avarvand A, Derakhshan M, Meshkat Z, Tafaghodi M, Ghazvini K, et al. Mycobacterium tuberculosis HspX/EsxS fusion protein: gene cloning, protein expression, and purification in Escherichia coli. Rep Biochem Mol Biol 2017;6:15-21.
17. Khademi F, Sahebkar A, Fasihi‐Ramandi M, Taheri RA. Induction of strong immune response against a multicomponent antigen of Mycobacterium tuberculosis in BALB/c mice using PLGA and DOTAP adjuvant. APMIS 2018;126:509-514.
18. Khademi F, Yousefi A, Derakhshan M, Najafi A, Tafaghodi M. Enhancing immunogenicity of novel multistage subunit vaccine of Mycobacterium tuberculosis using PLGA: DDA hybrid nanoparticles and MPLA: Subcutaneous administration. Iran J Basic Med Sci 2019;22:893-900.
19. Wang X, Zhang J, Liang J, Zhang Y, Teng X, Yuan X, et al. Protection against Mycobacterium tuberculosis infection offered by a new multistage subunit vaccine correlates with increased number of IFN-γ+ IL-2+ CD4+ and IFN-γ+ CD8+ T cells. PLoS One 2015;10:e0122560.
20. Avarvand AY, Meshkat Z, Khademi F, Tafaghodi M. Immunogenicity of HspX/EsxS fusion protein of Mycobacterium tuberculosis along with ISCOMATRIX and PLUSCOM nano-adjuvants after subcutaneous administration in animal model. Microb Pathog 2021;154:104842.
21. McBurney WT, Lendemans DG, Myschik J, Hennessy T, Rades T, Hook S. In vivo activity of cationic immune stimulating complexes (PLUSCOMs). Vaccine. 2008;26:4549-4556.
22. Baz Morelli A, Becher D, Koernig S, Silva A, Drane D, Maraskovsky E. ISCOMATRIX: A novel adjuvant for use in prophylactic and therapeutic vaccines against infectious diseases. J Med Microbiol 2012;61:935-943.
23. Lendemans DG, Myschik J, Hook S, Rades T. Cationic cage-like complexes formed by DC-cholesterol, Quil-A, and phospholipid. J Pharm Sci 2005;94:1794-1807.
24. Khademi F, Derakhshan M, Yousefi-Avarvand A, Najafi A, Tafaghodi M. A novel antigen of Mycobacterium tuberculosis and MPLA adjuvant co-entrapped into PLGA: DDA hybrid nanoparticles stimulates mucosal and systemic immunity. Microb Pathog 2018;125:507-513.
25. Drane D, Gittleson C, Boyle J, Maraskovsky E. ISCOMATRIX™ adjuvant for prophylactic and therapeutic vaccines. Expert Rev Vaccines 2007;6:761-772.
26. Maraskovsky E, Schnurr M, Wilson NS, Robson NC, Boyle J, Drane D. Development of prophylactic and therapeutic vaccines using the ISCOMATRIX adjuvant. Immunol Cell Biol 2009;87:371-376.
27. Bigaeva E, Doorn Ev, Liu H, Hak E. Meta-analysis on randomized controlled trials of vaccines with QS-21 or ISCOMATRIX adjuvant: Safety and tolerability. PLoS One 2016;11:e0154757.
28. Mehravaran A, Jaafari MR, Jalali SA, Khamesipour A, Tafaghodi M, Hojatizade M, et al. Cationic immune stimulating complexes containing soluble Leishmania antigens: Preparation, characterization and in vivo immune response evaluation. Iran J Immunol 2015;12:274-287.
29. Badiee A, Shargh VH, Khamesipour A, Jaafari MR. Micro/nanoparticle adjuvants for antileishmanial vaccines: Present and future trends. Vaccine 2013;31:735-749.
30. Tafaghodi M, Khademi F, Firouzi Z. Polymer-based nanoparticles as delivery systems for treatment and vaccination of tuberculosis.  Nanotechnology Based Approaches for Tuberculosis Treatment: Elsevier; 2020. p. 123-42.
31. Kaufmann SH, editor Tuberculosis vaccines: Time to think about the next generation. Semin Immunol 2013; 25:172-181.