The Role of ExoS in Dissemination of during Pneumonia

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Dissemination to the bloodstream is a poor prognostic sign in patients with hospital-acquired pneumonia, yet the mechanism by which this occurs is poorly understood. To begin to address this issue, we have used a mouse model of P. aeruginosa pneumonia to study the mechanism by which the type-III-secreted effector protein ExoS enhances bacterial dissemination. We show that intoxication of type I pneumocytes by ExoS leads to cell death and disruption of the pulmonary-vascular barrier, allowing bacterial dissemination into the bloodstream. These effects required the ADP-ribosyltransferase activity of ExoS, as strains secreting an ExoS variant lacking this activity demonstrated reduced type I pneumocytes death and pulmonary-vascular breakdown. This study indicates that inhibitors of the ADP-ribosyltransferase activity of ExoS could serve as novel therapeutics for the prevention of bacteremic pneumonia.

Vyšlo v časopise: The Role of ExoS in Dissemination of during Pneumonia. PLoS Pathog 11(6): e32767. doi:10.1371/journal.ppat.1004945
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004945

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1. Heyland DK, Cook DJ, Griffith L, Keenan SP, Brun-Buisson C (1999) The attributable morbidity and mortality of ventilator-associated pneumonia in the critically ill patient. The Canadian Critical Trials Group. Am J Respir Crit Care Med 159: 1249–1256. 10194173

2. Magret M, Lisboa T, Martin-Loeches I, Manez R, Nauwynck M, et al. (2011) Bacteremia is an independent risk factor for mortality in nosocomial pneumonia: a prospective and observational multicenter study. Crit Care 15: R62. doi: 10.1186/cc10036 21324159

3. Fagon JY, Chastre J, Domart Y, Trouillet JL, Pierre J, et al. (1989) Nosocomial pneumonia in patients receiving continuous mechanical ventilation. Am Rev Respir Dis 139: 877–884. 2930067

4. George DL, Galk PS, Wunderink RG, Leeper KVJ, Meduri GU, et al. (1998) Epidemiology of ventilator-acquired pneumonia based on protected bronchoscopic sampling. Am J Respir Crit Care Med 158: 1839–1847. 9847276

5. Rello J, Gallego M, Mariscal D, Sonora R, Valles J (1997) The value of routine microbial investigation in ventilator-associated pneumonia. Am J Respir Crit Care Med 156: 196–200. 9230747

6. Iannini PB, Claffey T, Quintiliani R (1974) Bacteremic Pseudomonas pneumonia. JAMA 230: 558–561. 4212965

7. Planquette B, Timsit JF, Misset BY, Schwebel C, Azoulay E, et al. (2013) Pseudomonas aeruginosa ventilator-associated pneumonia. predictive factors of treatment failure. Am J Respir Crit Care Med 188: 69–76. doi: 10.1164/rccm.201210-1897OC 23641973

8. Hauser AR (2009) The type III secretion system of Pseudomonas aeruginosa: infection by injection. Nat Rev Microbiol 7: 654–665. doi: 10.1038/nrmicro2199 19680249

9. Roy-Burman A, Savel RH, Racine S, Swanson BL, Revadigar NS, et al. (2001) Type III protein secretion is associated with death in lower respiratory and systemic Pseudomonas aeruginosa infections. J Infect Dis 183: 1767–1774. 11372029

10. Hauser AR, Cobb E, Bodí M, Mariscal D, Vallés J, et al. (2002) Type III protein secretion is associated with poor clinical outcomes in patients with ventilator-associated pneumonia caused by Pseudomonas aeruginosa. Crit Care Med 30: 521–528. 11990909

11. Feltman H, Schulert G, Khan S, Jain M, Peterson L, et al. (2001) Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa. Microbiology 147: 2659–2669. 11577145

12. Frithz-Lindsten E, Du Y, Rosqvist R, Forsberg A (1997) Intracellular targeting of exoenzyme S of Pseudomonas aeruginosa via type III-dependent translocation induces phagocytosis resistance, cytotoxicity and disruption of actin microfilaments. Mol Microbiol 25: 1125–1139. 9350868

13. Fleiszig SM, Wiener-Kronish JP, Miyazaki H, Vallas V, Mostov KE, et al. (1997) Pseudomonas aeruginosa-mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect Immun 65: 579–586. 9009316

14. Jia J, Wang Y, Zhou L, Jin S (2006) Expression of Pseudomonas aeruginosa toxin ExoS effectively induces apoptosis in host cells. Infect Immun 74: 6557–6570. 16966406

15. Kaufman MR, Jia J, Zeng L, Ha U, Chow M, et al. (2000) Pseudomonas aeruginosa mediated apoptosis requires the ADP-ribosylating activity of ExoS. Microbiology 146: 2531–2541. 11021928

16. Angus AA, Lee AA, Augustin DK, Lee EJ, Evans DJ, et al. (2008) Pseudomonas aeruginosa induces membrane blebs in epithelial cells, which are utilized as a niche for intracellular replication and motility. Infect Immun 76: 1992–2001. doi: 10.1128/IAI.01221-07 18316391

17. Kudoh I, Wiener-Kronish JP, Hashimoto S, Pittet J-F, Frank D (1994) Exoproduct secretions of P. aeruginosa strains influence severity of alveolar epithelial injury. Am J Physiol 267(5 Pt 1): L551–L556. 7977765

18. Shaver CM, Hauser AR (2004) Relative contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to virulence in the lung. Infect Immun 72: 6969–6977. 15557619

19. Nicas TI, Bradley J, Lochner JE, Iglewski BH (1985) The role of exoenzyme S in infections with Pseudomonas aeruginosa. J Infect Dis 152: 716–721. 2995500

20. Beasley MB, Travis W.D., Rubin E. (2008) The Respiratory System. In: Rubin R. Strayer DS, editor. Rubin's Pathology: Clinicopathologic. 5th ed. Philadelphia, PA: Lippincott Williams & Wilkins.

21. McCormack FX, Whitsett JA (2002) The pulmonary collectins, SP-A and SP-D, orchestrate innate immunity in the lung. J Clin Invest 109: 707–712. 11901176

22. Rangel SM, Logan LK, Hauser AR (2014) The ADP-ribosyltransferase domain of the effector protein ExoS inhibits phagocytosis of Pseudomonas aeruginosa during pneumonia. MBio 5: e01080–01014. doi: 10.1128/mBio.01080-14 24917597

23. Zlokarnik G, Negulescu PA, Knapp TE, Mere L, Burres N, et al. (1998) Quantitation of transcription and clonal selection of single living cells with beta-lactamase as reporter. Science 279: 84–88. 9417030

24. Verove J, Bernarde C, Bohn YS, Boulay F, Rabiet MJ, et al. (2012) Injection of Pseudomonas aeruginosa Exo toxins into host cells can be modulated by host factors at the level of translocon assembly and/or activity. PLoS One 7: e30488. doi: 10.1371/journal.pone.0030488 22299042

25. Charpentier X, Oswald E (2004) Identification of the secretion and translocation domain of the enteropathogenic and enterohemorrhagic Escherichia coli effector Cif, using TEM-1 beta-lactamase as a new fluorescence-based reporter. J Bacteriol 186: 5486–5495. 15292151

26. Marketon MM, DePaolo RW, DeBord KL, Jabri B, Schneewind O (2005) Plague bacteria target immune cells during infection. Science 309: 1739–1741. 16051750

27. Goehring U-M, Schmidt G, Pederson KJ, Aktories K, Barbieri JT (1999) The N-terminal domain of Pseudomonas aeruginosa exoenzyme S is a GTPase-activating protein for Rho GTPases. J Biol Chem 274: 36369–36372. 10593930

28. Radke J, Pederson KJ, Barbieri JT (1999) Pseudomonas aeruginosa exoenzyme S is a biglutamic acid ADP-ribosyltransferase. Infect Immun 67: 1508–1510. 10024602

29. Yahr TL, Barbieri JT, Frank DW (1996) Genetic relationship between the 53- and 49-kilodalton forms of exoenzyme S from Pseudomonas aeruginosa. J Bacteriol 178: 1412–1419. 8631719

30. Sun J, Barbieri JT (2003) Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 regulator of kinase (Crk) proteins. J Biol Chem 278: 32794–32800. 12807879

31. Shaver CM, Hauser AR (2006) Interactions between effector proteins of the Pseudomonas aeruginosa type III secretion system do not significantly affect several measures of disease severity in mammals. Microbiology 152: 143–152. 16385124

32. Bruno TF, Woods DE, Mody CH (2000) Exoenzyme S from Pseudomonas aeruginosa induces apoptosis in T lymphocytes. J Leukoc Biol 67: 808–816. 10857853

33. Vance RE, Rietsch A, Mekalanos JJ (2005) Role of the type III secreted exoenzymes S, T, and Y in systemic spread of Pseudomonas aeruginosa PAO1 in vivo. Infect Immun 73: 1706–1713. 15731071

34. Diaz MH, Shaver CM, King JD, Musunuri S, Kazzaz JA, et al. (2008) Pseudomonas aeruginosa induces localized immunosuppression during pneumonia in mammals. Infect Immun 76: 4414–4421. doi: 10.1128/IAI.00012-08 18663007

35. Tran CS, Rangel SM, Almblad H, Kierbel A, Givskov M, et al. (2014) The Pseudomonas aeruginosa type III translocon is required for biofilm formation at the epithelial barrier. PLoS Pathog 10: e1004479. doi: 10.1371/journal.ppat.1004479 25375398

36. Heiniger RW, Winther-Larsen HC, Pickles RJ, Koomey M, Wolfgang MC (2010) Infection of human mucosal tissue by Pseudomonas aeruginosa requires sequential and mutually dependent virulence factors and a novel pilus-associated adhesin. Cell Microbiol 12: 1158–1173. doi: 10.1111/j.1462-5822.2010.01461.x 20331639

37. Apodaca G, Bomsel M, Lindstedt R, Engel J, Frank D, et al. (1995) Characterization of Pseudomonas aeruginosa-induced MDCK cell injury: glycosylation defective host cells are resistant to bacterial killing. Infect Immun 63: 1541–1551. 7890421

38. Woodworth BA, Tamashiro E, Bhargave G, Cohen NA, Palmer JN (2008) An in vitro model of Pseudomonas aeruginosa biofilms on viable airway epithelial cell monolayers. Am J Rhinol 22: 235–238. doi: 10.2500/ajr.2008.22.3178 18588754

39. Soong G, Parker D, Magargee M, Prince AS (2008) The type III toxins of Pseudomonas aeruginosa disrupt epithelial barrier function. J Bacteriol 190: 2814–2821. doi: 10.1128/JB.01567-07 18165298

40. Huber P, Bouillot S, Elsen S, Attree I (2014) Sequential inactivation of Rho GTPases and Lim kinase by Pseudomonas aeruginosa toxins ExoS and ExoT leads to endothelial monolayer breakdown. Cell Mol Life Sci 71: 1927–1941. doi: 10.1007/s00018-013-1451-9 23974244

41. Angus AA, Evans DJ, Barbieri JT, Fleiszig SM (2010) The ADP-ribosylation domain of Pseudomonas aeruginosa ExoS is required for membrane bleb niche formation and bacterial survival within epithelial cells. Infect Immun 78: 4500–4510. doi: 10.1128/IAI.00417-10 20732998

42. Maldonado-Arocho FJ, Green C, Fisher ML, Paczosa MK, Mecsas J (2013) Adhesins and host serum factors drive Yop translocation by yersinia into professional phagocytes during animal infection. PLoS Pathog 9: e1003415. doi: 10.1371/journal.ppat.1003415 23818844

43. Durand EA, Maldonado-Arocho FJ, Castillo C, Walsh RL, Mecsas J (2010) The presence of professional phagocytes dictates the number of host cells targeted for Yop translocation during infection. Cell Microbiol 12: 1064–1082. doi: 10.1111/j.1462-5822.2010.01451.x 20148898

44. Kuang Z, Hao Y, Walling BE, Jeffries JL, Ohman DE, et al. (2011) Pseudomonas aeruginosa elastase provides an escape from phagocytosis by degrading the pulmonary surfactant protein-A. PLoS One 6: e27091. doi: 10.1371/journal.pone.0027091 22069491

45. Rubio F, Cooley J, Accurso FJ, Remold-O'Donnell E (2004) Linkage of neutrophil serine proteases and decreased surfactant protein-A (SP-A) levels in inflammatory lung disease. Thorax 59: 318–323. 15047952

46. Fleiszig SM, Evans DJ, Do N, Vallas V, Shin S, et al. (1997) Epithelial cell polarity affects susceptibility to P. aeruginosa invasion and cytotoxicity. Infect Immun 65: 2861–2867. 9199460

47. Lee A, Chow D, Haus B, Tseng W, Evans D, et al. (1999) Airway epithelial tight junctions and binding and cytotoxicity of Pseudomonas aeruginosa. Am J Physiol 277: L204–217. 10409249

48. Ganter MT, Roux J, Su G, Lynch SV, Deutschman CS, et al. (2008) Role of small GPTases and {alpha}v{beta}5 integrin in P. aeruginosa-induced increase in lung permeability. Am J Respir Cell Mol Biol.

49. Matthay MA, Wiener-Kronish JP (1990) Intact epithelial barrier function is critical for the resolution of alveolar edema in humans. Am Rev Respir Dis 142: 1250–1257. 2252240

50. Akca O, Koltka K, Uzel S, Cakar N, Pembeci K, et al. (2000) Risk factors for early-onset, ventilator-associated pneumonia in critical care patients: selected multiresistant versus nonresistant bacteria. Anesthesiology 93: 638–645. 10969295

51. Lange M, Hamahata A, Enkhbaatar P, Esechie A, Connelly R, et al. (2008) Assessment of vascular permeability in an ovine model of acute lung injury and pneumonia-induced Pseudomonas aeruginosa sepsis. Crit Care Med 36: 1284–1289. doi: 10.1097/CCM.0b013e318169ef74 18379256

52. Ware LB, Matthay MA (2001) Alveolar fluid clearance is impaired in the majority of patients with acute lung injury and the acute respiratory distress syndrome. Am J Respir Crit Care Med 163: 1376–1383. 11371404

53. Ozer EA, Allen JP, Hauser AR (2014) Characterization of the core and accessory genomes of Pseudomonas aeruginosa using bioinformatic tools Spine and AGEnt. BMC Genomics 15: 737. doi: 10.1186/1471-2164-15-737 25168460

54. Diaz MH, Hauser AR (2010) Pseudomonas aeruginosa cytotoxin ExoU is injected into phagocytic cells during acute pneumonia. Infect Immun 78: 1447–1456. doi: 10.1128/IAI.01134-09 20100855

55. Yasmin L, Jansson AL, Panahandeh T, Palmer RH, Francis MS, et al. (2006) Delineation of exoenzyme S residues that mediate the interaction with 14-3-3 and its biological activity. FEBS J 273: 638–646. 16420486

56. Howell HA, Logan LK, Hauser AR (2013) Type III secretion of ExoU is critical during early Pseudomonas aeruginosa pneumonia. MBio 4: e00032–00013. doi: 10.1128/mBio.00032-13 23481600

57. Lee VT, Smith RS, Tummler B, Lory S (2005) Activities of Pseudomonas aeruginosa effectors secreted by the type III secretion system in vitro and during infection. Infect Immun 73: 1695–1705. 15731070

58. Vogel HJ, Bonner DM (1956) Acetylornithinase of Escherichia coli partial purification and some properties. J Biol Chem 218: 97–106. 13278318

59. Nicas TI, Iglewski BH (1984) Isolation and characterization of transposon-induced mutants of Pseudomonas aeruginosa deficient in production of exoenzyme S. Infect Immun 45: 470–474. 6086529

60. Hoang TT, Kutchma AJ, Becher A, Schweizer HP (2000) Integration-proficient plasmids for Pseudomonas aeruginosa: site- specific integration and use for engineering of reporter and expression strains. Plasmid 43: 59–72. 10610820

61. Comolli JC, Hauser AR, Waite L, Whitchurch CB, Mattick JS, et al. (1999) Pseudomonas aeruginosa gene products PilT and PilU are required for cytotoxicity in vitro and virulence in a mouse model of acute pneumonia. Infect Immun 67: 3625–3630. 10377148

62. Ecker RC, Rogojanu R, Streit M, Oesterreicher K, Steiner GE (2006) An improved method for discrimination of cell populations in tissue sections using microscopy-based multicolor tissue cytometry. Cytometry A 69: 119–123. 16479616

63. Ecker RC, Steiner GE (2004) Microscopy-based multicolor tissue cytometry at the single-cell level. Cytometry A 59: 182–190. 15170597