Arshid Yousefi Avarvand - Department of Laboratory Sciences, School of Allied Medical Sciences, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Zahra Meshkat - Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
farzad khademi - Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
Ehsan Aryan - Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Mojtaba Sankian - Immunobiochemistry laboratory, Immunology Research Center, Bu-Ali Research Institute, Mashhad, Iran
Mohsen Tafaghodi - 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 ۲ weeks. Three weeks after the final vaccination, IFN-γ, IL-۴. IL-۱۷, and TGF-β concentrations in the supernatant of cultured splenocytes of vaccinated mice as well as serum titers of IgG۱ and IgG۲a 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.

HspX/EsxS, ISCOMATRIX, MPLA, Mycobacterium tuberculosis, Nasal administration, PLUSCOM