Localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells and its stimula-tory effects on the expression of proinflammatory cytokines by THP-1 cells
10.3760/cma.j.issn.0254-5101.2013.12.009
- VernacularTitle:CPSIT＿0271的定位、表达时相及其诱导THP-1细胞产生促炎症因子
- Author:
Zhiqing LI
;
Liangzhuan LIU
;
Haiying WU
;
Jing PENG
;
Lili CHEN
;
Qingzhi HE
;
Yimou WU
- Publication Type:Journal Article
- Keywords:
Chlamydophila psittaci;
CPSIT_0271;
Cellular localization;
Temporal expression;
Proin-flammatory cytokines
- From:Chinese Journal of Microbiology and Immunology
2013;(12):933-937
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the intracellular localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells; and to investigate the effects of recombinant GST-CPSIT_0271 protein on the expression of proinflammatory cytokines including IL-6, IL-1βand TNF-αby THP-1 cells.Methods The gene encoding CPSIT_0271 of Chlamydia psittaci was expressed as fusion protein ( GST-CPSIT_0271 ) in E.coli.The polyclonal antibody was prepared by immunizing BALB /c mice with the purified recombinant pro-tein.Antibody titer was determined by ELISA .Indirect immunofluorescence assay ( IFA) was performed to lo-cate the endogenous CPSIT_0271 protein in C.psittaci-infected cells .The expression characteristics of CPSIT_0271 protein were detected by Western blot in C.psittaci-infected HeLa cells at different time points .The levels of IL-6, IL-1βand TNF-αwere analyzed by ELISA after stimulating THP-1 cells with different concentrations of CPSIT_0271 protein.Results CPSIT_0271 protein was found to express in the chlamydia inclusion of C.psittaci-infected HeLa cells .The expression of CPSIT_0271 protein was detected firstly at 36 h and increased at 48 h after C.psittaci infection.The titer of anti-CPSIT_0271 specific antibody in GST-CPSIT_0271 immu-nized mice reached to 1 ∶16 000.GST-CPSIT_0271 protein increased the levels of IL-6, IL-1βand TNF-αin THP-1 cells in a dose-dependent manner in the range of 2 to 5 μg/ml.The levels of TNF-αand IL-1βreached their peaks at 24 h, and IL-6 level peaked at 48 h upon the stimulation by 5 μg/ml of GST-CPSIT_0271 pro-tein.Conclusion CPSIT_0271 expressed in inclusion bodies of Chlamydia psittaci in the infected cells , sug-gesting it might be a late expression gene .GST-CPSIT_0271 protein shows good immunogenicity and enhances the expressions of IL-6, IL-1βand TNF-αin THP-1 cells.
