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Published on: March 24, 2017
[Immunological characteristics of Mycobacterium tuberculosis antigen Rv2628]
Abstract:
Antigen Rv2628 of Mycobacterium tuberculosis is associated with latent tuberculosis infection. In this study, Rv2628 was prokaryotic expressed and purified, its immunological characteristics was evaluated with macrophage cell line RAW264.7 and BALB/c mice. The results show that Rv2628 was mainly expressed in form of inclusion body confirmed by SDS-PAGE, and could react with rabbit anti-H37Rv polyclonal antibody detected by Western blotting assay, indicating that the protein had an effective immunoreactivity. The interactions between Rv2628 and macrophage cell line RAW264.7 confirmed that it could effectively induce cells to produce pro-inflammatory cytokines, the relative expression level of IL-6 mRNA was higher than the control group in 1-12 h. BALB/c mice were subcutaneously immunized with Rv2628 protein, the production of IFN-gamma and IL-4 in the spleen cells was determined by Sandwich ELISA, in the Rv2628 immunized group, the level of IFN-gamma was significantly higher than that of IL-4 (P < 0.000 1). It indicated the protein induced Th1-tendency immune responses. At the same time, Rv2628(11-30) peptide used as coating antigen, the murine serum antibody titer detected by indirect-ELISA was 1:1 600, which demonstrated that Rv2628 could also induce humoral immune responses. In summary, Rv2628 could induce specific pro-inflammatory cytokines, affectively induce strongly Th1-tendency immune response and humoral response, it could be a potential target for developing subunit vaccine against TB. In addition, it laid foundation for probing the cross-talk between M. tb and host.
Insights
Mycobacterium tuberculosis antigen Rv2628 induces pro-inflammatory cytokines and Th1-biased immune responses. This antigen shows potential as a target for developing subunit vaccines against tuberculosis (TB).
Area of Science:
- Immunology
- Microbiology
- Vaccinology
Background:
- Antigen Rv2628 from Mycobacterium tuberculosis is linked to latent tuberculosis infection.
- Understanding the immunological role of Rv2628 is crucial for developing novel TB vaccines.
Purpose of the Study:
- To characterize the immunological properties of Mycobacterium tuberculosis antigen Rv2628.
- To evaluate its potential as a target for a subunit vaccine against tuberculosis.
Main Methods:
- Prokaryotic expression and purification of Rv2628.
- Immunological assays using macrophage cell line RAW264.7 and BALB/c mice.
- SDS-PAGE, Western blotting, cytokine analysis (IL-6 mRNA, IFN-gamma, IL-4), and antibody titer determination (indirect-ELISA).
Main Results:
- Rv2628 protein exhibited immunoreactivity and induced pro-inflammatory cytokines (IL-6).
- Immunization with Rv2628 induced a significant Th1-biased immune response (IFN-gamma > IL-4).
- Rv2628 also elicited a humoral immune response with a high antibody titer.
Conclusions:
- Rv2628 induces specific pro-inflammatory cytokines, Th1-biased immunity, and humoral responses.
- Rv2628 is a potential candidate antigen for developing a subunit vaccine against tuberculosis.
- This study provides a foundation for investigating host-pathogen interactions in M. tuberculosis infections.
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