1887

Abstract

Purpose. Previous research demonstrated that IL-10 was up-regulated in Chlamydia trachomatis-infected cells and that exogenous IL-10 was able to inhibit the secretion of pro-inflammatory cytokines by infected cells. However, the mechanisms are not well understood. The aim of this study was to investigate the mechanisms for up-regulation of IL-10 and inhibition of pro-inflammatory cytokine secretion in C. trachomatis-stimulated peripheral blood mononuclear cells (PBMCs).

Methodology. Human PBMCs were isolated from the blood of healthy human donors by standard Ficoll–Hypaque density gradient centrifugation. Cells were exposed to C. trachomatis in the presence or absence of MEK inhibitor U0126, the p38 inhibitor SB203580, the STAT3 inhibitor Ruxolitinib or anti-human IL-10 antibody. Cytokines were measured from culture supernatants using ELISA kits. Cells were harvested for real-time quantitative PCR to determine IL-10 mRNA levels and for Western blot assay to detect the expression of ERK1/2, p-ERK1/2, p38, p-p38, STAT3 and p-STAT3.

Results. Both mRNA and protein levels of IL-10 were up-regulated in stimulated cells, and the production of IL-10 was reduced when cells were treated with U0126 or SB203580. The expression of cytokines IL-6, IL-8 and TNF-α was enhanced in stimulated cells treated with anti-human IL-10 antibody. Moreover, neutralization of IL-10 resulted in a significant decrease of phosphorylated STAT3 in stimulated cells. Ruxolitinib caused a significant increase in the production of IL-6, IL-8 and TNF-α in stimulated cells.

Conclusion. IL-10 is up-regulated in an ERK- and p38-dependent fashion in stimulated human PBMCs. IL-10 inhibits the production of pro-inflammatory cytokines by activating the JAK/STAT signalling pathway.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000672
2018-01-08
2019-10-22
Loading full text...

Full text loading...

/deliver/fulltext/jmm/67/2/240.html?itemId=/content/journal/jmm/10.1099/jmm.0.000672&mimeType=html&fmt=ahah

References

  1. Hackstadt T. The diverse habitats of obligate intracellular parasites. Curr Opin Microbiol 1998; 1: 82– 87 [CrossRef] [PubMed]
    [Google Scholar]
  2. Hackstadt T, Fischer ER, Scidmore MA, Rockey DD, Heinzen RA. Origins and functions of the chlamydial inclusion. Trends Microbiol 1997; 5: 288– 293 [CrossRef] [PubMed]
    [Google Scholar]
  3. Hybiske K, Stephens RS. Mechanisms of host cell exit by the intracellular bacterium Chlamydia. Proc Natl Acad Sci USA 2007; 104: 11430– 11435 [CrossRef] [PubMed]
    [Google Scholar]
  4. Taylor HR, Johnson SL, Schachter J, Caldwell HD, Prendergast RA. Pathogenesis of trachoma: the stimulus for inflammation. J Immunol 1987; 138: 3023– 3027 [PubMed]
    [Google Scholar]
  5. Sherman KJ, Daling JR, Stergachis A, Weiss NS, Foy HM et al. Sexually transmitted diseases and tubal pregnancy. Sex Transm Dis 1990; 17: 115– 121 [CrossRef] [PubMed]
    [Google Scholar]
  6. Kuo CC, Grayston JT, Campbell LA, Goo YA, Wissler RW et al. Chlamydia pneumoniae (TWAR) in coronary arteries of young adults (15–34 years old). Proc Natl Acad Sci USA 1995; 92: 6911– 6914 [CrossRef] [PubMed]
    [Google Scholar]
  7. Campbell LA, Kuo CC. Chlamydia pneumoniae and atherosclerosis. Semin Respir Infect 2003; 18: 48– 54 [CrossRef] [PubMed]
    [Google Scholar]
  8. Skwor TA, Atik B, Kandel RP, Adhikari HK, Sharma B et al. Role of secreted conjunctival mucosal cytokine and chemokine proteins in different stages of trachomatous disease. PLoS Negl Trop Dis 2008; 2: e264 [CrossRef] [PubMed]
    [Google Scholar]
  9. Dessus-Babus S, Darville TL, Cuozzo FP, Ferguson K, Wyrick PB. Differences in innate immune responses (in vitro) to HeLa cells infected with nondisseminating serovar E and disseminating serovar L2 of Chlamydia trachomatis. Infect Immun 2002; 70: 3234– 3248 [CrossRef] [PubMed]
    [Google Scholar]
  10. Ji J, Sahu GK, Braciale VL, Cloyd MW. HIV-1 induces IL-10 production in human monocytes via a CD4-independent pathway. Int Immunol 2005; 17: 729– 736 [CrossRef] [PubMed]
    [Google Scholar]
  11. Sloan DD, Jerome KR. Herpes simplex virus remodels T-cell receptor signaling, resulting in p38-dependent selective synthesis of interleukin-10. J Virol 2007; 81: 12504– 12514 [CrossRef] [PubMed]
    [Google Scholar]
  12. Kekarainen T, Montoya M, Mateu E, Segalés J. Porcine circovirus type 2-induced interleukin-10 modulates recall antigen responses. J Gen Virol 2008; 89: 760– 765 [CrossRef] [PubMed]
    [Google Scholar]
  13. Balcewicz-Sablinska MK, Gan H, Remold HG. Interleukin 10 produced by macrophages inoculated with Mycobacterium avium attenuates mycobacteria-induced apoptosis by reduction of TNF-α activity. J Infect Dis 1999; 180: 1230– 1237 [CrossRef] [PubMed]
    [Google Scholar]
  14. Sabat R. IL-10 family of cytokines. Cytokine Growth Factor Rev 2010; 21: 315– 324 [CrossRef] [PubMed]
    [Google Scholar]
  15. Filippi CM, von Herrath MG. IL-10 and the resolution of infections. J Pathol 2008; 214: 224– 230 [CrossRef] [PubMed]
    [Google Scholar]
  16. Ohman H, Tiitinen A, Halttunen M, Birkelund S, Christiansen G et al. IL-10 polymorphism and cell-mediated immune response to Chlamydia trachomatis. Genes Immun 2006; 7: 243– 249 [CrossRef] [PubMed]
    [Google Scholar]
  17. Iyer SS, Cheng G. Role of interleukin 10 transcriptional regulation in inflammation and autoimmune disease. Crit Rev Immunol 2012; 32: 23– 63 [CrossRef] [PubMed]
    [Google Scholar]
  18. Wills-Karp M, Nathan A, Page K, Karp CL. New insights into innate immune mechanisms underlying allergenicity. Mucosal Immunol 2010; 3: 104– 110 [CrossRef] [PubMed]
    [Google Scholar]
  19. Kinnunen A, Surcel HM, Halttunen M, Tiitinen A, Morrison RP et al. Chlamydia trachomatis heat shock protein-60 induced interferon-γ and interleukin-10 production in infertile women. Clin Exp Immunol 2003; 131: 299– 303 [CrossRef] [PubMed]
    [Google Scholar]
  20. Geng Y, Shane RB, Berencsi K, Gonczol E, Zaki MH et al. Chlamydia pneumoniae inhibits apoptosis in human peripheral blood mononuclear cells through induction of IL-10. J Immunol 2000; 164: 5522– 5529 [CrossRef] [PubMed]
    [Google Scholar]
  21. Yilma AN, Singh SR, Fairley SJ, Taha MA, Dennis VA. The anti-inflammatory cytokine, interleukin-10, inhibits inflammatory mediators in human epithelial cells and mouse macrophages exposed to live and UV-inactivated Chlamydia trachomatis. Mediators Inflamm 2012; 2012: 1– 10 [CrossRef] [PubMed]
    [Google Scholar]
  22. Kun D, Xiang-Lin C, Ming Z, Qi L. Chlamydia inhibit host cell apoptosis by inducing Bag-1 via the MAPK/ERK survival pathway. Apoptosis 2013; 18: 1083– 1092 [CrossRef] [PubMed]
    [Google Scholar]
  23. Gao Q, Qi L, Wu T, Wang J. An important role of interleukin-10 in counteracting excessive immune response in HT-29 cells exposed to Clostridium butyricum. BMC Microbiol 2012; 12: 100 [CrossRef] [PubMed]
    [Google Scholar]
  24. Yi AK, Yoon JG, Yeo SJ, Hong SC, English BK et al. Role of mitogen-activated protein kinases in CpG DNA-mediated IL-10 and IL-12 production: central role of extracellular signal-regulated kinase in the negative feedback loop of the CpG DNA-mediated Th1 response. J Immunol 2002; 168: 4711– 4720 [PubMed] [Crossref]
    [Google Scholar]
  25. Chi H, Barry SP, Roth RJ, Wu JJ, Jones EA et al. Dynamic regulation of pro- and anti-inflammatory cytokines by MAPK phosphatase 1 (MKP-1) in innate immune responses. Proc Natl Acad Sci USA 2006; 103: 2274– 2279 [CrossRef] [PubMed]
    [Google Scholar]
  26. Du K, Zheng Q, Zhou M, Zhu L, Ai B et al. Chlamydial antiapoptotic activity involves activation of the Raf/MEK/ERK survival pathway. Curr Microbiol 2011; 63: 341– 346 [CrossRef] [PubMed]
    [Google Scholar]
  27. Yilma AN, Singh SR, Morici L, Dennis VA. Flavonoid naringenin: a potential immunomodulator for Chlamydia trachomatis inflammation. Mediators Inflamm 2013; 2013: 1– 13 [CrossRef] [PubMed]
    [Google Scholar]
  28. Hakimi H, Zare-Bidaki M, Zainodini N, Assar S, Arababadi MK. Significant roles played by IL-10 in Chlamydia infections. Inflammation 2014; 37: 818– 823 [CrossRef] [PubMed]
    [Google Scholar]
  29. Batteiger BE, Xu F, Johnson RE, Rekart ML. Protective immunity to Chlamydia trachomatis genital infection: evidence from human studies. J Infect Dis 2010; 201: 178– 189 [CrossRef] [PubMed]
    [Google Scholar]
  30. Azenabor AA, York J. Chlamydia trachomatis evokes a relative anti-inflammatory response in a free Ca2+ dependent manner in human macrophages. Comp Immunol Microbiol Infect Dis 2010; 33: 513– 528 [CrossRef] [PubMed]
    [Google Scholar]
  31. Ohman H, Tiitinen A, Halttunen M, Lehtinen M, Paavonen J et al. Cytokine polymorphisms and severity of tubal damage in women with Chlamydia-associated infertility. J Infect Dis 2009; 199: 1353– 1359 [CrossRef] [PubMed]
    [Google Scholar]
  32. Moniz RJ, Chan AM, Kelly KA. Identification of dendritic cell subsets responding to genital infection by Chlamydia muridarum. FEMS Immunol Med Microbiol 2009; 55: 226– 236 [CrossRef] [PubMed]
    [Google Scholar]
  33. Rizzo A, Paolillo R, Lanza AG, Guida L, Annunziata M et al. Chlamydia pneumoniae induces interleukin-6 and interleukin-10 in human gingival fibroblasts. Microbiol Immunol 2008; 52: 447– 454 [CrossRef] [PubMed]
    [Google Scholar]
  34. Igietseme JU, Ananaba GA, Bolier J, Bowers S, Moore T et al. Suppression of endogenous IL-10 gene expression in dendritic cells enhances antigen presentation for specific Th1 induction: potential for cellular vaccine development. J Immunol 2000; 164: 4212– 4219 [CrossRef] [PubMed]
    [Google Scholar]
  35. Yang X, Gartner J, Zhu L, Wang S, Brunham RC. IL-10 gene knockout mice show enhanced Th1-like protective immunity and absent granuloma formation following Chlamydia trachomatis lung infection. J Immunol 1999; 162: 1010– 1017 [PubMed]
    [Google Scholar]
  36. Hou J, Wang L, Quan R, Fu Y, Zhang H et al. Induction of interleukin-10 is dependent on p38 mitogen-activated protein kinase pathway in macrophages infected with porcine reproductive and respiratory syndrome virus. Virol J 2012; 9: 165 [CrossRef] [PubMed]
    [Google Scholar]
  37. Su H, McClarty G, Dong F, Hatch GM, Pan ZK et al. Activation of Raf/MEK/ERK/cPLA2 signaling pathway is essential for chlamydial acquisition of host glycerophospholipids. J Biol Chem 2004; 279: 9409– 9416 [CrossRef] [PubMed]
    [Google Scholar]
  38. Bea F, Puolakkainen MH, McMillen T, Hudson FN, Mackman N et al. Chlamydia pneumoniae induces tissue factor expression in mouse macrophages via activation of Egr-1 and the MEK-ERK1/2 pathway. Circ Res 2003; 92: 394– 401 [CrossRef] [PubMed]
    [Google Scholar]
  39. Lad SP, Li J, da Silva Correia J, Pan Q, Gadwal S et al. Cleavage of p65/RelA of the NF-kappaB pathway by Chlamydia. Proc Natl Acad Sci USA 2007; 104: 2933– 2938 [CrossRef] [PubMed]
    [Google Scholar]
  40. Buchholz KR, Stephens RS. The extracellular signal-regulated kinase/mitogen-activated protein kinase pathway induces the inflammatory factor interleukin-8 following Chlamydia trachomatis infection. Infect Immun 2007; 75: 5924– 5929 [CrossRef] [PubMed]
    [Google Scholar]
  41. Sun Y, Zhou P, Chen S, Hu C, Bai Q et al. The JAK/STAT3 signaling pathway mediates inhibition of host cell apoptosis by Chlamydia psittaci infection. Pathog Dis 2017; [CrossRef] [PubMed]
    [Google Scholar]
  42. Sellami H, Said-Sadier N, Znazen A, Gdoura R, Ojcius DM et al. Chlamydia trachomatis infection increases the expression of inflammatory tumorigenic cytokines and chemokines as well as components of the Toll-like receptor and NF-κB pathways in human prostate epithelial cells. Mol Cell Probes 2014; 28: 147– 154 [CrossRef] [PubMed]
    [Google Scholar]
  43. Yu P, Xiao L, Lin L, Tang L, Chen C et al. STAT3-mediated TLR2/4 pathway upregulation in an IFN-γ-induced Chlamydia trachomatis persistent infection model. Pathog Dis 2016; 74: ftw076 [CrossRef]
    [Google Scholar]
  44. Takagi H, Sanada T, Minoda Y, Yoshimura A. Regulation of cytokine and toll-like receptor signaling by SOCS family genes. Nihon Rinsho 2004; 62: 2189– 2196 [PubMed]
    [Google Scholar]
  45. Penttilä T, Haveri A, Tammiruusu A, Vuola JM, Lahesmaa R et al. Chlamydia pneumoniae infection in IL-10 knock out mice: accelerated clearance but severe pulmonary inflammatory response. Microb Pathog 2008; 45: 25– 29 [CrossRef] [PubMed]
    [Google Scholar]
  46. Omosun Y, Mckeithen D, Ryans K, Kibakaya C, Blas-Machado U et al. Interleukin-10 modulates antigen presentation by dendritic cells through regulation of NLRP3 inflammasome assembly during Chlamydia infection. Infect Immun 2015; 83: 4662– 4672 [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000672
Loading
/content/journal/jmm/10.1099/jmm.0.000672
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error