Recombinant Cpn 0810 stimulates proinflammatory cytokine expression and apoptosis in human monocytes

Chen,Yuyu; Wu,Baiping; Liu,Liangzhuan; You,Xiaoxing; Chen,Lili; Wu,Yimou; Zhang,Qiugui

doi:10.3892/etm.2014.2111

Recombinant Cpn 0810 stimulates proinflammatory cytokine expression and apoptosis in human monocytes

Authors:
- Yuyu Chen
- Baiping Wu
- Liangzhuan Liu
- Xiaoxing You
- Lili Chen
- Yimou Wu
- Qiugui Zhang
View Affiliations

Affiliations: Department of Inspection, The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, Hunan 410013, P.R. China, Department of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, P.R. China, Department of Inspection, The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, P.R. China
Published online on: December 5, 2014 https://doi.org/10.3892/etm.2014.2111
Pages: 459-463

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to express the recombinant Chlamydophila pneumoniae (C. pneumoniae) protein, Cpn 0810, in Escherichia coli (E. coli) BL21, and investigate the effects of Cpn 0810 on inflammatory and apoptotic processes in human monocytic (THP‑1) cells. An ELISA was performed to detect the levels of the proinflammatory cytokines, tumor necrosis factor (TNF)‑α and interleukin (IL)‑6. In addition, Hoechst 33258 staining and annexin V binding analyses were performed to measure the rates of apoptosis. Purified glutathione S‑transferase (GST)‑Cpn 0810 recombinant proteins were obtained from the E. coli BL21 cells carrying the pGEX6p‑2/Cpn 0810 plasmid, and were shown to stimulate the expression of TNF‑α and IL‑6 in the THP‑1 cells in a dose‑ and time‑dependent manner. TNF‑α and IL‑6 levels peaked at 24 h after GST‑Cpn 0810 stimulation. Furthermore, GST‑Cpn 0810 significantly promoted the apoptosis of THP‑1 cells. In conclusion, recombinant GST‑Cpn 0810 was shown to stimulate the expression of TNF‑α and IL‑6, inhibit proliferation and induce apoptosis in THP‑1 cells. Therefore, Cpn 0810 may interact with host cells following C. pneumoniae infection, functioning as an important pathogenic factor.

View Figures

View References

1	Lui G, Ip M, Lee N, et al: Role of ‘atypical pathogens’ among adult hospitalized patients with community-acquired pneumonia. Respirology. 14:1098–1105. 2009. View Article : Google Scholar : PubMed/NCBI
2	Damy SB, Higuchi ML, Timenetsky J, et al: Mycoplasma pneumoniae and/or Chlamydophila pneumoniae inoculation causing different aggravations in cholesterol-induced atherosclerosis in apoE KO male mice. BMC Microbiol. 9:1942009. View Article : Google Scholar : PubMed/NCBI
3	Papaetis GS, Anastasakou E and Orphanidou D: Chlamydophila pneumoniae infection and COPD: more evidence for lack of evidence? Eur J Intern Med. 20:579–585. 2009. View Article : Google Scholar : PubMed/NCBI
4	Jupelli M, Shimada K, Chiba N, et al: Chlamydia pneumoniae infection in mice induces chronic lung inflammation, iBALT formation, and fibrosis. PLoS One. 8:e774472013. View Article : Google Scholar : PubMed/NCBI
5	Mota LJ and Cornelis GR: The bacterial injection kit: type III secretion systems. Ann Med. 37:234–249. 2005. View Article : Google Scholar : PubMed/NCBI
6	Mota LJ, Sorg I and Cornelis GR: Type III secretion: the bacteria-eukaryotic cell express. FEMS Microbiol Lett. 252:1–10. 2005. View Article : Google Scholar : PubMed/NCBI
7	Journet L, Hughes KT and Cornelis GR: Type III secretion: a secretory pathway serving both motility and virulence (review). Mol Membr Biol. 22:41–50. 2005. View Article : Google Scholar : PubMed/NCBI
8	Müller N, Sattelmacher F, Lugert R and Gross U: Characterization and intracellular localization of putative Chlamydia pneumoniae effector proteins. Med Microbiol Immunol. 197:387–396. 2008. View Article : Google Scholar : PubMed/NCBI
9	Stone CB, Johnson DL, Bulir DC, Gilchrist JD and Mahony JB: Characterization of the putative type III secretion ATPase CdsN (Cpn0707) of Chlamydophila pneumoniae. J Bacteriol. 190:6580–6588. 2008. View Article : Google Scholar : PubMed/NCBI
10	Herrmann M, Schuhmacher A, Mühldorfer I, Melchers K, Prothmann C and Dammeier S: Identification and characterization of secreted effector proteins of Chlamydophila pneumoniae TW183. Res Microbiol. 157:513–524. 2006. View Article : Google Scholar : PubMed/NCBI
11	Cortes C, Rzomp KA, Tvinnereim A, Scidmore MA and Wizel B: Chlamydia pneumoniae inclusion membrane protein Cpn0585 interacts with multiple Rab GTPases. Infect Immun. 75:5586–5596. 2007. View Article : Google Scholar : PubMed/NCBI
12	Luo J, Jia T, Zhong Y, Chen D, Flores R and Zhong G: Localization of the hypothetical protein Cpn0585 in the inclusion membrane of Chlamydia pneumoniae-infected cells. Microb Pathog. 42:111–116. 2007. View Article : Google Scholar : PubMed/NCBI
13	Kariagina AS, Alekseevskiĭ AV, Spirin SA, Zigangirova NA and Gintsburg AL: Effector proteins of Clamidia. Mol Biol (Mosk). 43:963–983. 2009.(In Russian).
14	Luo J, Jia T, Flores R, Chen D and Zhong G: Hypothetical protein Cpn0308 is localized in the Chlamydia pneumoniae inclusion membrane. Infect Immun. 75:497–503. 2007. View Article : Google Scholar :
15	Flores R, Luo J, Chen D, Sturgeon G, Shivshankar P, Zhong Y and Zhong G: Characterization of the hypothetical protein Cpn1027, a newly identified inclusion membrane protein unique to Chlamydia pneumoniae. Microbiology. 153:777–786. 2007. View Article : Google Scholar : PubMed/NCBI
16	Fields KA and Hackstadt T: Evidence for the secretion of Chlamydia trachomatis CopN by a type III secretion mechanism. Mol Microbiol. 38:1048–1060. 2000. View Article : Google Scholar : PubMed/NCBI
17	Lugert R, Kuhns M, Polch T and Gross U: Expression and localization of type III secretion-related proteins of Chlamydia pneumoniae. Med Microbiol Immunol. 193:163–171. 2004. View Article : Google Scholar
18	Beeckman DS, Geens T, Timmermans JP, Van Oostveldt P and Vanrompay DC: Identification and characterization of a type III secretion system in Chlamydophila psittaci. Vet Res. 39:272008. View Article : Google Scholar : PubMed/NCBI
19	Jamison WP and Hackstadt T: Induction of type III secretion by cell-free Chlamydia trachomatis elementary bodies. Microb Pathog. 45:435–440. 2008. View Article : Google Scholar : PubMed/NCBI
20	Häcker H and Karin M: Regulation and function of IKK and IKK-related kinases. Sci STKE. 2006:re132006. View Article : Google Scholar : PubMed/NCBI
21	Pfeffer K: Biological functions of tumor necrosis factor cytokines and their receptors. Cytokine Growth Factor Rev. 14:185–191. 2003. View Article : Google Scholar : PubMed/NCBI
22	Perrins CJ and Bobryshev YV: Current advances in understanding of immunopathology of atherosclerosis. Virchows Arch. 458:117–123. 2011. View Article : Google Scholar
23	Hu S, Liang S, Guo H, et al: Comparison of the inhibition mechanisms of adalimumab and infliximab in treating tumor necrosis factor α-associated diseases from a molecular view. J Biol Chem. 288:27059–27067. 2013. View Article : Google Scholar : PubMed/NCBI
24	Paladino N, Mul Fedele ML, Duhart JM, Marpegan L and Golombek DA: Modulation of mammalian circadian rhythms by tumor necrosis factor-α. Chronobiol Int. 1–12. 2014.
25	Li L, Li DH, Qu N, Wen WM and Huang WQ: The role of ERK1/2 signaling pathway in coronary microembolization-induced rat myocardial inflammation and injury. Cardiology. 117:207–215. 2010. View Article : Google Scholar : PubMed/NCBI
26	Tisato V, Zauli G, Rimondi E, et al: Inhibitory effect of natural anti-inflammatory compounds on cytokines released by chronic venous disease patient-derived endothelial cells. Mediators Inflamm. 2013:4234072013.
27	Johnston SC, Zhang H, Messina LM, Lawton MT and Dean D: Chlamydia pneumoniae burden in carotid arteries is associated with upregulation of plaque interleukin-6 and elevated C-reactive protein in serum. Arterioscler Thromb Vasc Biol. 25:2648–2653. 2005. View Article : Google Scholar : PubMed/NCBI
28	Ridker PM, Rifai N, Stampfer MJ and Hennekens CH: Plasma concentration of interleukin-6 and the risk of future myocardial infarction among apparently healthy men. Circulation. 101:1767–1772. 2000. View Article : Google Scholar : PubMed/NCBI
29	Sharma J, Dong F, Pirbhai M and Zhong G: Inhibition of proteolytic activity of a chlamydial proteasome/protease-like activity factor by antibodies from humans infected with Chlamydia trachomatis. Infect Immun. 73:4414–4419. 2005. View Article : Google Scholar : PubMed/NCBI
30	Biasucci LM, Santamaria M and Liuzzo G: Inflammation, atherosclerosis and acute coronary syndromes. Minerva Cardioangiol. 50:475–486. 2002.(In Italian). PubMed/NCBI
31	Geng YJ and Libby P: Evidence for apoptosis in advanced human atheroma. Colocalization with interleukin-1 beta-converting enzyme. Am J Pathol. 147:251–266. 1995.PubMed/NCBI
32	Hayakawa Y, Takemura G, Misao J, et al: Apoptosis and overexpression of bax protein and bax mRNA in smooth muscle cells within intimal hyperplasia of human radial arteries: analysis with arteriovenous fistulas used for hemodialysis. Arterioscler Thromb Vasc Biol. 19:2066–2077. 1999. View Article : Google Scholar : PubMed/NCBI