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Medical Microbiology and Immunology

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01.08.2011 | Original Investigation

Analysis of Chlamydia pneumoniae infection in mononuclear cells by reverse transcription-PCR targeted to chlamydial gene transcripts

verfasst von: Laura Mannonen, Eveliina Markkula, Mirja Puolakkainen

Erschienen in: Medical Microbiology and Immunology | Ausgabe 3/2011

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Abstract

Chlamydia pneumoniae (C. pneumoniae) is an important etiological agent of respiratory infections including pneumonia. C. pneumoniae DNA can be detected in peripheral blood mononuclear cells indicating that monocytes can assist the spread of infection to other anatomical sites. Persistent infection established at these sites could promote inflammation and enhance pathology. Thus, the mononuclear cells are in a strategic position in the development of persistent infection. To investigate the intracellular replication and fate of C. pneumoniae in mononuclear cells, we have established an in vitro model in the human Mono Mac 6 cell line. In the present study, we analyzed the transcription of 11 C. pneumoniae genes in Mono Mac 6 cells during infection by real-time RT-PCR. Our results suggest that the transcriptional profile of the studied genes in monocytes is different from that seen in epithelial cells. Furthermore, our study shows that genes related to secretion are transcribed, and secreted bacterial proteins are also translated during infection of monocytes, creating novel opportunities for the management of chlamydial infection of monocytes.

Haider S, Wagner M, Schmid MC, Sixt BS, Christian JG, Häcker G, Pichler P, Mechtler K, Müller A, Baranyi C, Toenshoff ER, Montanaro J, Horn M (2010) Raman microspectroscopy reveals long-term extracellular activity of chlamydiae. Mol Microbiol 77:687–700PubMedCrossRef

Saikku P (1992) The epidemiology and significance of Chlamydia pneumoniae. J Infect 25(Suppl 1):27–34PubMedCrossRef

Campbell LA, Kuo CC (2004) Chlamydia pneumoniae—an infectious risk factor for atherosclerosis? Nat Rev Microbiol 2:23–32PubMedCrossRef

Hahn DL, Dodge RW, Golubjatnikov R (1991) Association of Chlamydia pneumoniae (strain TWAR) infection with wheezing, asthmatic bronchitis, and adult-onset asthma. JAMA 266:225–230PubMedCrossRef

Saikku P, Leinonen M, Mattila K, Ekman MR, Nieminen MS, Mäkelä PH, Huttunen JK, Valtonen V (1988) Serological evidence of an association of a novel Chlamydia, TWAR, with chronic coronary heart disease and acute myocardial infarction. Lancet 2:983–986PubMedCrossRef

Beagley KW, Huston WM, Hansbro PM, Timms P (2009) Chlamydial infection of immune cells: altered function and implications for disease. Crit Rev Immunol 29:275–305PubMed

Boman J, Söderberg S, Forsberg J, Birgander LS, Allard A, Persson K, Jidell E, Kumlin U, Juto P, Waldenström A, Wadell G (1998) High prevalence of Chlamydia pneumoniae DNA in peripheral blood mononuclear cells in patients with cardiovascular disease and in middle-aged blood donors. J Infect Dis 178:274–277PubMedCrossRef

Maass M, Jahn J, Gieffers J, Dalhoff K, Katus HA, Solbach W (2000) Detection of Chlamydia pneumoniae within peripheral blood monocytes of patients with unstable angina or myocardial infarction. J Infect Dis 3(Suppl):S449–S451CrossRef

West SK, Kohlhepp SJ, Jin R, Gleaves CA, Stamm W, Gilbert DN (2009) Detection of circulating Chlamydophila pneumoniae in patients with coronary artery disease and healthy control subjects. Clin Infect Dis 48(5):560–567PubMedCrossRef

10.

Epstein SE, Zhu J, Burnett MS, Zhou YF, Vercellotti G, Hajjar D (2000) Infection and atherosclerosis: potential roles of pathogen burden and molecular mimicry. Arterioscler Thromb Vasc Biol 20:1417–1420PubMedCrossRef

11.

Lehr HA, Sagban TA, Ihling C, Zähringer U, Hungerer KD, Blumrich M, Reifenberg K, Bhakdi S (2001) Immunopathogenesis of atherosclerosis: endotoxin accelerates atherosclerosis in rabbits on hypercholesterolemic diet. Circulation 104:914–920PubMedCrossRef

12.

Bhakdi S, Lackner KJ, Han SR, Torzewski M, Husmann M (2004) Beyond cholesterol: the enigma of atherosclerosis revisited. Thromb Haemost 91:639–645PubMed

13.

Byrne GI, Ouellette SP, Wang Z, Rao JP, Lu L, Beatty WL, Hudson AP (2001) Chlamydia pneumoniae expresses genes required for DNA replication but not cytokinesis during persistent infection of HEp-2 cells. Infect Immun 69:5423–5429PubMedCrossRef

14.

Mannonen L, Kamping E, Penttilä T, Puolakkainen M (2004) IFN-γ induced persistent Chlamydia pneumoniae infection in HL and Mono Mac 6 cells: characterization by real-time quantitative PCR and culture. Microb Pathog 36:41–50PubMedCrossRef

15.

Bobryshev YV, Killingsworth MC, Tran D, Lord R (2008) Amalgamation of Chlamydia pneumoniae inclusions with lipid droplets in foam cells in human atherosclerotic plaque. Virchows Arch 453:69–77PubMedCrossRef

16.

Al-Younes HM, Rudel T, Brinkmann V, Szczepek AJ, Meyer TF (2001) Low iron availability modulates the course of Chlamydia pneumoniae infection. Cell Microbiol 3:427–437PubMedCrossRef

17.

Beatty WL, Byrne GI, Morrison RP (1993) Morphologic and antigenic characterization of interferon γ-mediated persistent Chlamydia trachomatis infection in vitro. Proc Natl Acad Sci USA 90:3998–4002PubMedCrossRef

18.

Clark RB, Schatzki PF, Dalton HP (1982) Ultrastructural analysis of the effects of erythromycin on the morphology and developmental cycle of Chlamydia trachomatis HAR-13. Arch Microbiol 133:278–282PubMedCrossRef

19.

Coles AM, Reynolds DJ, Harper A, Devitt A, Pearce JH (1993) Low-nutrient induction of abnormal chlamydial development: a novel component of chlamydial pathogenesis? FEMS Microbiol Lett 106:193–200PubMedCrossRef

20.

Kutlin A, Roblin PM, Hammerschlag MR (1999) In vitro activities of azithromycin and ofloxacin against Chlamydia pneumoniae in a continuous-infection model. Antimicrob Agents Chemother 43:2268–2272PubMed

21.

Moulder JW, Levy NJ, Schulman LP (1980) Persistent infection of mouse fibroblasts (L cells) with Chlamydia psittaci: evidence for a cryptic chlamydial form. Infect Immun 30:874–883PubMed

22.

Heinemann M, Susa M, Simnacher U, Marre R, Essig A (1996) Growth of Chlamydia pneumoniae induces cytokine production and expression of CD14 in a human monocytic cell line. Infect Immun 64:4872–4875PubMed

23.

Wahl C, Oswald F, Simnacher U, Weiss S, Marre R, Essig A (2001) Survival of Chlamydia pneumoniae-infected Mono Mac 6 cells is dependent on NF-κB binding activity. Infect Immun 69:7039–7045PubMedCrossRef

24.

Numazaki K, Chiba S (1996) Serum γ-interferon in patients with pneumonia caused by Chlamydia pneumoniae. Pediatr Infect Dis J 15:174–175PubMedCrossRef

25.

Penttilä JM, Anttila M, Puolakkainen M, Laurila A, Varkila K, Sarvas M, Mäkelä PH, Rautonen N (1998) Local immune responses to Chlamydia pneumoniae in the lungs of BALB/c mice during primary infection and reinfection. Infect Immun 66:5113–5118PubMed

26.

Mathews S, George C, Flegg C, Stenzel D, Timms P (2001) Differential expression of ompA, ompB, pyk, nlpD and Cpn0585 genes between normal and interferon-γ treated cultures of Chlamydia pneumoniae. Microb Pathog 30:337–345PubMedCrossRef

27.

Mäurer AP, Mehlitz A, Mollenkopf HJ, Meyer TF (2007) Gene expression profiles of Chlamydophila pneumoniae during the developmental cycle and iron depletion-mediated persistence. PLoS Pathog 3:e83PubMedCrossRef

28.

Ouellette SP, Hatch TP, AbdelRahman YM, Rose LA, Belland RJ, Byrne GI (2006) Global transcriptional upregulation in the absence of increased translation in Chlamydia during IFNγ-mediated host cell tryptophan starvation. Mol Microbiol 62:1387–1401PubMedCrossRef

29.

Polkinghorne A, Hogan RJ, Vaughan L, Summersgill JT, Timms P (2005) Differential expression of chlamydial signal transduction genes in normal and interferon γ-induced persistent Chlamydophila pneumoniae infections. Microbes Infect 8:61–72PubMedCrossRef

30.

Slepenkin A, Motin V, de la Maza LM, Peterson EM (2003) Temporal expression of type III secretion genes of Chlamydia pneumoniae. Infect Immun 71:2555–2562PubMedCrossRef

31.

Timms P, Good D, Wan C, Theodoropoulos C, Mukhopadhyay S, Summersgill J, Mathews S (2009) Differential transcriptional responses between the interferon-gamma-induction and iron-limitation models of persistence for Chlamydia pneumoniae. J Microbiol Immunol Infect 42:27–37PubMed

32.

Airenne S, Surcel HM, Alakärppä H, Laitinen K, Paavonen J, Saikku P, Laurila A (1999) Chlamydia pneumoniae infection in human monocytes. Infect Immun 67:1445–1449PubMed

33.

Gieffers J, Füllgraf H, Jahn J, Klinger M, Dalhoff K, Katus HA, Solbach W, Maass M (2001) Chlamydia pneumoniae infection in circulating human monocytes is refractory to antibiotic treatment. Circulation 103:351–356PubMed

34.

Haranaga S, Yamaguchi H, Ikejima H, Friedman H, Yamamoto Y (2003) Chlamydia pneumoniae infection of alveolar macrophages: a model. J Infect Dis 187:1107–1115PubMedCrossRef

35.

Klos A, Thalmann J, Peters J, Gérard HC, Hudson AP (2009) The transcript profile of persistent Chlamydophila (Chlamydia) pneumoniae in vitro depends on the means by which persistence is induced. FEMS Microbiol Lett 29:120–126CrossRef

36.

Hsia RC, Pannekoek Y, Ingerowski E, Bavoil PM (1997) Type III secretion genes identify a putative virulence locus of Chlamydia. Mol Microbiol 25:351–359PubMedCrossRef

37.

Stephens RS, Kalman S, Lammel C, Fan J, Marathe R, Aravind L, Mitchell W, Olinger L, Tatusov RL, Zhao Q, Koonin EV, Davis RW (1998) Genome sequence of an obligate intracellular pathogen of humans: Chlamydia trachomatis. Science 282:754–759PubMedCrossRef

38.

Clifton DR, Fields KA, Grieshaber SS, Dooley CA, Fischer ER, Mead DJ, Carabeo RA, Hackstadt T (2004) A chlamydial type III translocated protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. Proc Natl Acad Sci USA 101:10166–10171PubMedCrossRef

39.

Lugert R, Kuhns M, Polch T, Gross U (2004) Expression and localization of type III secretion-related proteins of Chlamydia pneumoniae. Med Microbiol Immunol 193:163–171PubMedCrossRef

40.

Müller N, Sattelmacher F, Lugert R, Gross U (2008) Characterization and intracellular localization of putative Chlamydia pneumoniae effector proteins. Med Microbiol Immunol 197:387–396PubMedCrossRef

41.

Subtil A, Delevoye C, Balañá ME, Tastevin L, Perrinet S, Dautry-Varsat A (2005) A directed screen for chlamydial proteins secreted by a type III mechanism identifies a translocated protein and numerous other new candidates. Mol Microbiol 56:1636–1647PubMedCrossRef

42.

Slepenkin A, Enquist PA, Hägglund U, de la Maza LM, Elofsson M, Peterson EM (2007) Reversal of the antichlamydial activity of putative type III secretion inhibitors by iron. Infect Immun 75:3478–3489PubMedCrossRef

43.

Cles LD, Stamm WE (1990) Use of HL cells for improved isolation and passage of Chlamydia pneumoniae. J Clin Microbiol 28:938–940PubMed

44.

Ekman MR, Grayston JT, Visakorpi R, Kleemola M, Kuo CC, Saikku P (1993) An epidemic of infections due to Chlamydia pneumoniae in military conscripts. Clin Infect Dis 17:420–425PubMedCrossRef

45.

Summersgill JT, Sahney NN, Gaydos CA, Quinn TC, Ramirez JA (1995) Inhibition of Chlamydia pneumoniae growth in HEp-2 cells pretreated with gamma interferon and tumor necrosis factor alpha. Infect Immun 63:2801–2803PubMed

46.

Mannonen L, Nikula T, Haveri A, Reinikainen A, Vuola JM, Lahesmaa R, Puolakkainen M (2007) Up-regulation of host cell genes during interferon-γ-induced persistent Chlamydia pneumoniae infection in HL cells. J Infect Dis 195:212–219PubMedCrossRef

47.

Airaksinen U, Penttilä T, Wahlström E, Vuola JM, Puolakkainen M, Sarvas M (2003) Production of Chlamydia pneumoniae proteins in Bacillus subtilis and their use in characterizing immune responses in the experimental infection model. Clin Diagn Lab Immunol 10:367–375PubMed

48.

Hefty PS, Stephens RS (2007) Chlamydial type III secretion system is encoded on ten operons preceded by sigma 70-like promoter elements. J Bacteriol 189:198–206PubMedCrossRef

49.

Hogan RJ, Mathews SA, Mukhopadhyay S, Summersgill JT, Timms P (2004) Chlamydial persistence: beyond the biphasic paradigm. Infect Immun 72:1843–1855PubMedCrossRef

50.

Bannantine JP, Griffiths RS, Viratyosin W, Brown WJ, Rockey DD (2000) A secondary structure motif predictive of protein localization to the chlamydial inclusion membrane. Cell Microbiol 2:35–47PubMedCrossRef

51.

Hackstadt T, Fischer ER, Scidmore MA, Rockey DD, Heinzen RA (1997) Origins and functions of the chlamydial inclusion. Trends Microbiol 5:288–293PubMedCrossRef

52.

Hackstadt T, Scidmore-Carlson MA, Shaw EI, Fischer ER (1999) The Chlamydia trachomatis IncA protein is required for homotypic vesicle fusion. Cell Microbiol 1:119–130PubMedCrossRef

53.

Paumet F, Wesolowski J, Garcia-Diaz A, Delevoye C, Aulner N, Shuman HA, Subtil A, Rothman JE (2009) Intracellular bacteria encode inhibitory SNARE-like proteins. PLoS ONE 4:e7375PubMedCrossRef

54.

Wyrick PB (2000) Intracellular survival by Chlamydia. Cell Microbiol 2:275–282PubMedCrossRef

55.

Watson MW, Clarke IN, Everson JS, Lambden PR (1995) The CrP operon of Chlamydia psittaci and Chlamydia pneumoniae. Microbiology 141:2489–2497PubMedCrossRef

56.

Heuer D, Brinkmann V, Meyer TF, Szczepek AJ (2003) Expression and translocation of chlamydial protease during acute and persistent infection of the epithelial HEp-2 cells with Chlamydophila (Chlamydia) pneumoniae. Cell Microbiol 5:315–322PubMedCrossRef

57.

Dong F, Su H, Huang Y, Zhong Y, Zhong G (2004) Cleavage of host keratin 8 by a Chlamydia-secreted protease. Infect Immun 72:3863–3868PubMedCrossRef

58.

Kumar Y, Valdivia RH (2008) Actin and intermediate filaments stabilize the Chlamydia trachomatis vacuole by forming dynamic structural scaffolds. Cell Host Microbe 4:159–169PubMedCrossRef

59.

Hogan RJ, Mathews SA, Kutlin A, Hammerschlag MR, Timms P (2003) Differential expression of genes encoding membrane proteins between acute and continuous Chlamydia pneumoniae infections. Microb Pathog 34:11–16PubMedCrossRef

60.

Nicholson TL, Olinger L, Chong K, Schoolnik G, Stephens RS (2003) Global stage-specific gene regulation during the developmental cycle of Chlamydia trachomatis. J Bacteriol 185:3179–3189PubMedCrossRef

61.

Gérard HC, Köhler L, Branigan PJ, Zeidler H, Schumacher HR, Hudson AP (1998) Viability and gene expression in Chlamydia trachomatis during persistent infection of cultured human monocytes. Med Microbiol Immunol 187:115–120PubMedCrossRef

62.

Koehler L, Nettelnbreker E, Hudson AP, Ott N, Gérard HC, Branigan PJ, Schumacher HR, Drommer W, Zeidler H (1997) Ultrastructural and molecular analyses of the persistence of Chlamydia trachomatis (serovar K) in human monocytes. Microb Pathog 22:133–142PubMedCrossRef

63.

Schmitz E, Nettelnbreker E, Zeidler H, Hammer M, Manor E, Wollenhaupt J (1993) Intracellular persistence of chlamydial major outer-membrane protein, lipopolysaccharide and ribosomal RNA after non-productive infection of human monocytes with Chlamydia trachomatis serovar K. J Med Microbiol 38:278–285PubMedCrossRef

64.

Bailey L, Gylfe A, Sundin C, Muschiol S, Elofsson M, Nordström P, Henriques-Normark B, Lugert R, Waldenström A, Wolf-Watz H, Bergström S (2007) Small molecule inhibitors of type III secretion in Yersinia block the Chlamydia pneumoniae infection cycle. FEBS Lett 581:587–595PubMedCrossRef

Titel: Analysis of Chlamydia pneumoniae infection in mononuclear cells by reverse transcription-PCR targeted to chlamydial gene transcripts
verfasst von: Laura Mannonen
Eveliina Markkula
Mirja Puolakkainen
Publikationsdatum: 01.08.2011
Verlag: Springer-Verlag
Erschienen in: Medical Microbiology and Immunology / Ausgabe 3/2011
Print ISSN: 0300-8584
Elektronische ISSN: 1432-1831
DOI: https://doi.org/10.1007/s00430-011-0184-3

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Analysis of Chlamydia pneumoniae infection in mononuclear cells by reverse transcription-PCR targeted to chlamydial gene transcripts

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