Open Access

Recombinant Bacille Calmette-Guérin coexpressing Ag85B-IFN-γ enhances the cell-mediated immunity in C57BL/6 mice

  • Authors:
    • Wei Liu
    • Ying Xu
    • Hongbo Shen
    • Jingran Yan
    • Enzhuo Yang
    • Honghai Wang
  • View Affiliations

  • Published online on: March 28, 2017     https://doi.org/10.3892/etm.2017.4273
  • Pages: 2339-2347
  • Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The only available vaccine against pulmonary tuberculosis is Bacille Calmette‑Guérin (BCG). As the efficacy reported of the vaccine is not up to the mark, there is an urgent need to develop improved anti‑tuberculosis vaccines. Antigen 85B (Ag85B) is a very promising vaccine candidate molecule of Mycobacterium tuberculosis and interferon (IFN)‑γ and has been considered the most attractive correlate of protective immunity. The aim of this study was to construct a novel recombinant BCG (rBCG) to secrete Ag85B and mouse IFN‑γ under control of the Mycobacterial heat shock protein 60 (hsp60) promoter and the antigen signal sequence. Second aim of the present study is to evaluate the immune response in C57BL/6 elicted by the new rBCG. Expression of the fusion protein was readily detectable by western blotting and IFN‑γ bioactivity was detected indirectly by enzyme‑linked immunosorbent assay (ELISA). Compared with BCG, rBCG::Ag85B‑IFN‑γ was substantially more active in inducing the production of IFN‑γ and tumor necrosis factor (TNF)‑α from mouse splenocytes. ELISA analysis for IgG, IgG1 and IgG2c showed that rBCG::Ag85B‑IFN‑γ induced higher titer of Ag85B and facilitated Th1 type immune response. rBCG::Ag85B‑IFN‑γ also improved nitric oxide production levels and enhanced antigen-specific splenocyte proliferation. Moreover, rBCG::Ag85B‑IFN‑γ induced human monocytes such as THP‑1 cells to enhance expression of CD80, CD86, CD40 and HLA‑DR. Flow cytometry analysis confirmed that rBCG::Ag85B-IFN-γ significantly activated CD4+  T cells. Assessing combinations of IFN‑γ, TNF‑α and interleukin‑2 at the single‑cell level by multiparameter flow cytometry, we found that rBCG::Ag85B‑IFN‑γ improved the multifunctional T cells level in comparison to BCG. In conclusion, the present study indicates that rBCG::Ag85B‑IFN‑γ increases cell mediated immune response and is a potential candidate vaccine for immunotherapeutic protocols against pulmonary tuberculosis.
View Figures
View References

Related Articles

Journal Cover

May-2017
Volume 13 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Liu W, Xu Y, Shen H, Yan J, Yang E and Wang H: Recombinant Bacille Calmette-Guérin coexpressing Ag85B-IFN-γ enhances the cell-mediated immunity in C57BL/6 mice. Exp Ther Med 13: 2339-2347, 2017
APA
Liu, W., Xu, Y., Shen, H., Yan, J., Yang, E., & Wang, H. (2017). Recombinant Bacille Calmette-Guérin coexpressing Ag85B-IFN-γ enhances the cell-mediated immunity in C57BL/6 mice. Experimental and Therapeutic Medicine, 13, 2339-2347. https://doi.org/10.3892/etm.2017.4273
MLA
Liu, W., Xu, Y., Shen, H., Yan, J., Yang, E., Wang, H."Recombinant Bacille Calmette-Guérin coexpressing Ag85B-IFN-γ enhances the cell-mediated immunity in C57BL/6 mice". Experimental and Therapeutic Medicine 13.5 (2017): 2339-2347.
Chicago
Liu, W., Xu, Y., Shen, H., Yan, J., Yang, E., Wang, H."Recombinant Bacille Calmette-Guérin coexpressing Ag85B-IFN-γ enhances the cell-mediated immunity in C57BL/6 mice". Experimental and Therapeutic Medicine 13, no. 5 (2017): 2339-2347. https://doi.org/10.3892/etm.2017.4273