Study based on prokaryotic expression of Mycobacterium tuberculosis Rv2153c and evaluation of its immunogenicity
10.13200/j.cnki.cjb.004652
- VernacularTitle:基于结核分枝杆菌Rv2153c的原核表达及其免疫原性评价
- Author:
Manhong XUE
1
Author Information
1. Laboratory Department of Maternity Hospital Affiliated to Nanjing Medical University, Nanjing Women and Children’s Healthcare Hospital, Nanjing 210001, Jiangsu Province, China
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis(Mtb);
Rv2153c;
MurG;
Prokaryotic expression;
Multiplexed microsphere-based flow cytometric assay(MFCIA);
Immunogenicity
- From:
Chinese Journal of Biologicals
2026;39(02):137-143
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant vector for Mycobacterium tuberculosis(Mtb) Rv2153c(MurG), express it in prokaryotic cells, and then immunize C57 BL/6 mice to assess the immunogenicity.Methods The recombinant vector pProRv2153c was constructed, expressed in prokaryotic cells and purified. The purified product was identified by SDS-PAGE and Western blot, and the levels of related cytokines in peripheral blood of Chinese Mtb infected patients induced by recombinant Rv2153c protein were measured by multiplexed microsphere-based flow cytometric assay(MFCIA). The female C57 BL/6 mice were randomly divided into PBS, Rv2153c/SAS subunit vaccine, SAS and BCG groups with 6 mice in each group. After 9 weeks of immunization, the levels of specific antibodies in peripheral blood and secreted cytokines in splenocyte supernatant were detected.Results The recombinant vector pPro-Rv2153c was constructed correctly as identified by double enzyme digestion. The expressed Rv2153c protein had a relative molecular mass of about 43 200, and had specific reaction with Monoclonal Mouse Anti-His Tag and MurG Antibody. After stimulation with recombinant Rv2153c protein, the levels of IFNγ, TNF-α and IL-6 in subjects with active pulmonary tuberculosis(ATB) were significantly higher than those in latent tuberculosis infection(LTBI) and healthy control(HC) subjects. The serum specific antibody titers in Rv2153c/SAS group were significantly higher than those in the rest of the groups, and the ratio of IgG2a to IgG1 was greater than 1. In addition, the levels of secreted IFNγ, TNF-α and IL-2 in splenocyte supernatant of Rv2153c/SAS group were also significantly higher than those of other groups.Conclusion The recombinant Rv2153c protein can be specifically recognized by peripheral blood T cells of subjects infected with Mtb in China, and can induce strong antigen-specific Th1-type cellular immune response in the combined adjuvant SAS immunized mice, confirming the possible use of Rv2153c as a candidate target antigen for novel TB vaccine.
