Cloning, expression and biological activity of Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10
10.7644/j.issn.1674-9960.2016.03.015
- VernacularTitle:结核分枝杆菌ESAT-6＋CFP-10融合抗原克隆表达及其生物学活性
- Author:
Yanfeng ZHAO
;
Xiaoyan FENG
;
Bingshui XIU
;
Cuimi DUAN
;
Yawen FAN
;
Heqiu ZHANG
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
ESAT-6 and CFP-10 fusion antigen;
THP-1 cell;
cytokines
- From:
Military Medical Sciences
2016;40(3):226-229
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10 ( EC) and to evaluate the biological activity of the fusion antigen EC in inducing specific cytokines secretion from THP -1 cells. Methods The fusion antigen EC gene was cloned into pET-30 a prokaryotic expression vector and expressed highly in E.coli BL21.Then, the THP-1 cells were stimulated with purified fusion antigen EC of different concentrations (10 and 20 μg/ml).Culture supernatants were collected after 12 h and 24 h, respectively.The secretion levels of IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-αand IFN-γin THP-1 cell culture supernatants were detected using Bio-PlexProTM Assays kit.Results The M.tuberculosis fusion antigen EC was cloned and expressed successfully .The secretion levels of IL-6, IL-8 and TNF-αin EC infected THP-1 cells were significantly higher than those in THP-1 cells (P<0.05).The secretion levels of other cytokines did not change significantly .Conclusion The obtained M.tuberculosis fusion antigen EC has biological activity in inducing the THP-1 cells to secrete a higher level of IL-6,IL-8 and TNF-α.
