Recombinant Mycobacterium smegmatis Expressing an ESAT6-CFP10 Fusion Protein Induces Anti-Mycobacterial Immune Responses and Protects Against Mycobacterium tuberculosis Challenge in Mice

ABSTRACT Bioengineered bacterial polyester inclusions have the potential to be used as a vaccine delivery system. The biopolyester beads were engineered to display a fusion protein of the polyester synthase PhaC and the two key antigens involved in immune response to the infectious agent that causes tuberculosis, Mycobacterium tuberculosis, notably antigen 85A (Ag85A) and the 6-kDa early secreted antigenic target (ESAT-6) from Mycobacterium tuberculosis. Polyester beads displaying the respective fusion protein at a high density were successfully produced (henceforth called Ag85A-ESAT-6 beads) by recombinant Escherichia coli. The ability of the Ag85A-ESAT-6 beads to enhance mouse immunity to the displayed antigens was investigated. The beads were not toxic to the animals, as determined by weight gain and absence of lesions at the inoculation site in immunized animals. In vivo injection of the Ag85A-ESAT-6 beads in mice induced significant humoral and cell-mediated immune responses to both Ag85A and ESAT-6. Vaccination with Ag85A-ESAT-6 beads was efficient at stimulating immunity on their own, and this ability was enhanced by administration of the beads in an oil-in-water emulsion. In addition, vaccination with the Ag85A-ESAT-6 beads induced significantly stronger humoral and cell-mediated immune responses than vaccination with an equivalent dose of the fusion protein Ag85A-ESAT-6 alone. The immune response induced by the beads was of a mixed Th1/Th2 nature, as assessed from the induction of the cytokine gamma interferon (Th1 immune response) and increased levels of immunoglobulin G1 (Th2 immune response). Hence, engineered biopolyester beads displaying foreign antigens represent a new class of versatile, safe, and biocompatible vaccines.

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Corrigendum to “Immune responses to recombinant Mycobacterium smegmatis expressing fused core protein and preS1 peptide of hepatitis B virus in mice” [J. Virol. Methods 141 (2007) 41–48]

Journal of Virological Methods ◽

10.1016/j.jviromet.2007.05.019 ◽

2007 ◽

Vol 144 (1-2) ◽

pp. 172

Author(s):

Xingbin Hu ◽

Wen Yin ◽

Qiaohong Yue ◽

Xueqing Xu ◽

Sanhua Wei ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Immune Responses ◽

Mycobacterium Smegmatis ◽

Core Protein ◽

B Virus ◽

Recombinant Mycobacterium Smegmatis

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Prime-boost Mycobacterium smegmatis recombinant vaccination improves protection in mice infected with Mycobacterium tuberculosis by enhancing humoral and cellular specific immune responses

Frontiers in Immunology ◽

10.3389/conf.fimmu.2013.02.00068 ◽

2013 ◽

Vol 4 ◽

Author(s):

Junqueira-Kipnis Ana Paula ◽

Oliveira F�bio ◽

Trentini Monalisa ◽

Araujo-Filho Joao ◽

Kipnis Andr�

Keyword(s):

Mycobacterium Tuberculosis ◽

Immune Responses ◽

Mycobacterium Smegmatis

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

10.1101/2021.08.27.458002 ◽

2021 ◽

Author(s):

Arshid Yousefi Avarvand ◽

Zahra Meshkat ◽

Farzad Khademi ◽

Ehsan Aryan ◽

Mojtaba Sankian ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Immune Responses ◽

Fusion Protein ◽

Intranasal Administration ◽

Bacterial Infections ◽

Protective Efficacy ◽

Nasal Administration ◽

Mice Model ◽

Ifn Γ ◽

Nasal Immunization

Background: 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 tuberculosis 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 the intranasal administration in mice model. Methods:In 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. Finally, IFN-γ, IL-4. IL-17 and TGF-β concentration in 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 induced 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 immune response against of M. tuberculosis antigens. So, nasal immunization with these formulations, could induce immune responses and considered as new TB vaccine or as BCG booster.

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