Analysis of immunogenicity of tuberculosis fusion protein consisting of Ag85B, Mpt64 and HspX antigens expressed in replication and dormancy bacilli
10.3760/cma.j.issn.0254-5101.2009.02.002
- VernacularTitle:结核分枝杆菌抗原Ag85B-Mpt64190-198-HspX融合蛋白免疫原性的初步观察
- Author:
Qing LI
;
Wenwen JIANG
;
Yu LUO
;
Hongjuan YU
;
Nannan SONG
;
Bingxiang WANG
;
Xin LIU
;
Ying ZHANG
;
Bingdong ZHU
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
Subunit vaccine;
Fusion protein;
Ag85B;
HspX
- From:
Chinese Journal of Microbiology and Immunology
2009;29(2):103-107
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct protective immunity to Mycobncterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods Ag85B and Mpt64190-198-HspX sequences were amplified by PCR and cloned into plasmids pET-28a. The fusion protein, Ag85BMpt64190-198-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCGPSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64190-198 and HspX. Conclusion It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.
