The Steroid Catabolic Pathway of the Intracellular Pathogen Is Important for Pathogenesis and a Target for Vaccine Development
Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551), ipdB (rv3552), fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD) and 3aα-H-4α(3′-propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP). Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections.
Vyšlo v časopise:
The Steroid Catabolic Pathway of the Intracellular Pathogen Is Important for Pathogenesis and a Target for Vaccine Development. PLoS Pathog 7(8): e32767. doi:10.1371/journal.ppat.1002181
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002181
Souhrn
Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551), ipdB (rv3552), fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD) and 3aα-H-4α(3′-propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP). Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections.
Zdroje
1. PrescottJF 1991 Rhodococcus equi: an animal and human pathogen. Clin Microbiol Rev 4 20 34
2. MosserDMHondalusMK 1996 Rhodococcus equi: an emerging opportunistic pathogen. Trends Microbiol 4 29 33
3. YamshchikovAVSchuetzALyonGM 2010 Rhodococcus equi infection. Lancet Infect Dis 10 350 359
4. HondalusMKMosserDM 1994 Survival and replication of Rhodococcus equi in macrophages. Infect Immun 62 4167 4175
5. HondalusMK 1997 Pathogenesis and virulence of Rhodococcus equi. Vet Microbiol 56 257 268
6. MeijerWGPrescottJF 2004 Rhodococcus equi. Vet Res 35 383 396
7. von BargenKHaasA 2009 Molecular and infection biology of the horse pathogen Rhodococcus equi. FEMS Microbiol Rev 33 870 891
8. GiguèreSPrescottJF 1997 Clinical manifestations, diagnosis, treatment, and prevention of Rhodococcus equi infections in foals. Vet Microbiol 56 313 334
9. TakaiSSekizakiTOzawaTSugawaraTWatanabeY 1991 Association between a large plasmid and 15- to 17-kilodalton antigens in virulent Rhodococcus equi. Infect Immun 59 4056 4060
10. TakaiSHinesSASekizakiTNicholsonVMAlperinDA 2000 DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC33701 and 103. Infect Immun 68 6840 6847
11. GiguereSHondalusMKYagerJADarrahPMosserDM 1999 Role of the 85-kilobase plasmid and plasmid-encoded virulence-associated protein A in intracellular survival and virulence of Rhodococcus equi. Infect Immun 67 3548 3557
12. JainSBloomBRHondalusMK 2003 Deletion of vapA encoding Virulence Associated Protein A attenuates the intracellular actinomycete Rhodococcus equi. Mol Microbiol 50 115 128
13. LetekMOcampo-SosaAASandersMFogartyUBuckleyT 2008 Evolution of the Rhodococcus equi vap pathogenicity island seen through comparison of host-associated vapA and vapB virulence plasmids. J Bacteriol 190 5797 5805
14. Tkachuk-SaadOPrescottJF 1991 Rhodococcus equi plasmids: isolation and partial characterization. J Clin Microbiol 29 2696 2700
15. MajnoGJorisI 1995 Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146 3 15
16. LührmannAMauderNSydorTFernandez-MoraESchulze-LuehrmannJ 2004 Necrotic death of Rhodococcus equi-infected macrophages is regulated by virulence-associated plasmids. Infect Immun 72 853 862
17. von BargenKPolidoriMBeckenUHuthGPrescottJF 2009 Rhodococcus equi virulence-associated protein A is required for diversion of phagosome biogenesis but not for cytotoxicity. Infect Immun 77 5676 5681
18. CoulsonGBAgarwalSHondalusMK 2010 Characterization of the role of the pathogenicity island and vapG in the virulence of the intracellular actinomycete pathogen Rhodococcus equi. Infect Immun 78 3323 3334
19. OliveiraAFFerrazLCBrocchiMRoque-BarreiraMC 2007 Oral administration of a live attenuated Salmonella vaccine strain expressing the VapA protein induces protection against infection by Rhodococcus equi. Microbes Infect 9 382 390
20. OliveiraAFRuasLPCardosoSASoaresSGRoque-BarreiraMC 2010 Vaccination of mice with salmonella expressing VapA: mucosal and systemic Th1 responses provide protection against Rhodococcus equi infection. PLoS One 5 e8644
21. HaghighiHRPrescottJF 2005 Assessment in mice of vapA-DNA vaccination against Rhodococcus equi infection. Vet Immunol Immunopathol 104 215 225
22. PhumoonnaTBartonMDVanniasinkamTHeuzenroederMW 2008 Chimeric vapA/groEL2 DNA vaccines enhance clearance of Rhodococcus equi in aerosol challenged C3H/He mice. Vaccine 26 2457 2465
23. AshourJHondalusMK 2003 Phenotypic mutants of the intracellular actinomycete Rhodococcus equi created by in vivo Himar1 transposon mutagenesis. J Bacteriol 185 2644 2652
24. LopezAMTownsendHGAllenALHondalusMK 2008 Safety and immunogenicity of a live-attenuated auxotrophic candidate vaccine against the intracellular pathogen Rhodococcus equi. Vaccine 26 998 1009
25. NavasJGonzález-ZornBLadrónNGarridoPVázquez-BolandJA 2001 Identification and mutagenesis by allelic exchange of choE, encoding a cholesterol oxidase from the intracellular pathogen Rhodococcus equi. J Bacteriol 183 4796 4805
26. PeiYDupontCSydorTHaasAPrescottJF 2006 Cholesterol oxidase (ChoE) is not important in the virulence of Rhodococcus equi. Vet Microbiol 118 240 246
27. PeiYNicholsonVWoodsKPrescottJF 2007 Immunization by intrabronchial administration to 1-week-old foals of an unmarked double gene disruption strain of Rhodococcus equi strain 103+. Vet Microbiol 125 100 110
28. WallDMDuffyPSDupontCPrescottJFMeijerWG 2005 Isocitrate lyase activity is required for virulence of the intracellular pathogen Rhodococcus equi. Infect Immun 73 6736 6741
29. PeiYParreiraVNicholsonVMPrescottJF 2007 Mutation and virulence assessment of chromosomal genes of Rhodococcus equi 103. Can J Vet Res 71 1 7
30. HeHZahrtTC 2005 Identification and characterization of a regulatory sequence recognized by Mycobacterium tuberculosis persistence regulator MprA. J Bacteriol 187 202 212
31. MacArthurIParreiraVRLeppDMuthariaLMVazquez-BolandJA 2011 The sensor kinase MprB is required for Rhodococcus equi virulence. Vet Microbiol In press
32. AhmadSRoyPKBasuSKJohriBN 1993 Cholesterol side-chain cleavage by immobilized cells of Rhodococcus equi DSM89-133. Indian J Exp Biol 31 319 322
33. MurohisaTIidaM 1993 Some new intermediates in microbial side chain degradation of β-sitosterol. J Ferment Bioeng 76 174 177
34. Van der GeizeRde JongWHesselsGIGrommenAWJacobsAA 2008 A novel method to generate unmarked gene deletions in the intracellular pathogen Rhodococcus equi using 5-fluorocytosine conditional lethality. Nucleic Acids Res 36 e151
35. LeeSSSihCJ 1967 Mechanisms of steroid oxidation by microorganisms. XII. Metabolism of hexahydroindanpropionic acid derivatives. Biochemistry 6 1395 1403
36. Van der GeizeRYamKHeuserTWilbrinkMHHaraH 2007 A gene cluster encoding cholesterol catabolism in a soil actinomycete provides insight into Mycobacterium tuberculosis survival in macrophages. Proc Natl Acad Sci USA 104 1947 1952
37. PandeyAKSassettiCM 2008 Mycobacterial persistence requires the utilization of host cholesterol. Proc Natl Acad Sci USA 105 4376 4380
38. YamKCD′AngeloIKalscheuerRZhuHWangJX 2009 Studies of a ring-cleaving dioxygenase illuminate the role of cholesterol metabolism in the pathogenesis of Mycobacterium tuberculosis. PLoS Pathog 5 e1000344
39. HuYvan der GeizeRBesraGSGurchaSSLiuA 2010 3-Ketosteroid 9alpha-hydroxylase is an essential factor in the pathogenesis of Mycobacterium tuberculosis. Mol Microbiol 75 107 121
40. MohnWWvan der GeizeRStewartGROkamotoSLiuJ 2008 The actinobacterial mce4 locus encodes a steroid transporter. J Biol Chem 283 35368 35374
41. RengarajanJBloomBRRubinEJ 2005 Genome-wide requirements for Mycobacterium tuberculosis adaptation and survival in macrophages. Proc Natl Acad Sci USA 102 8327 8332
42. LetekMGonzálezPMacarthurIRodríguezHFreemanTC 2010 The genome of a pathogenic Rhodococcus: cooptive virulence underpinned by key gene acquisitions. PLoS Genet 6 e1001145
43. FinnRDMistryJTateJCoggillPHegerA 2010 The Pfam protein families database. Nucleic Acids Res Database Issue 38 D211 222
44. HorinouchiMYamamotoTTaguchiKAraiHKudoT 2001 Meta-cleavage enzyme tesB is necessary for testosterone degradation in Comamonas testosteroni TA441. Microbiology (UK) 147 3367 3375
45. HorinouchiMHayashiTKoshinoHKuritaTKudoT 2005 Identification of 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid, 4-hydroxy-2-oxohexanoic acid, and 2-hydroxyhexa-2,4-dienoic acid and related enzymes involved in testosterone degradation in Comamonas testosteroni TA441. Appl Environ Microbiol 71 5275 5281
46. MicloAGermainP 1990 Catabolism of methylperhydroindanedione propionate by Rhodococcus equi: evidence of a MEPHIP-reductase activity. Appl Microbiol Biotechnol 32 594 599
47. MicloAGermainP 1992 Hexahydroindanone derivatives of steroids formed by a Rhodococcus equi. Appl Microbiol Biotechnol 36 456 460
48. Van der GeizeRHesselsGIvan GerwenRvan der MeijdenPDijkhuizenL 2002 Molecular and functional characterization of kshA and kshB, encoding two components of 3-ketosteroid 9α-hydroxylase, a class IA monooxygenase, in Rhodococcus erythropolis strain SQ1. Mol Microbiol 45 1007 1018
49. LoriaRM 2009 Beta-androstenes and resistance to viral and bacterial infections. Neuroimmunomodulation 16 88 95
50. KendallSLBurgessPBalhanaRWithersMTen BokumA 2010 Cholesterol utilization in mycobacteria is controlled by two TetR-type transcriptional regulators: kstR and kstR2. Microbiology 156 1362 1371
51. Chirino-TrejoJMPrescottJF 1987 Antibody response of horses to Rhodococcus equi antigens. Can J Vet Res 51 301 305
52. TakaiSShodaMSasakiYTsubakiSFortierG 1999 Restriction fragment length polymorphisms of virulence plasmids in Rhodococcus equi. J Clin Microbiol. 37 3417 3420
53. Hooper-McGrevyKEWilkieBNPrescottJF 2005 Virulence-associated protein-specific serum immunoglobulin G-isotype expression in young foals protected against Rhodococcus equi pneumonia by oral immunization with virulent R. equi. Vaccine 23 5760 5767
54. MartensRJMartensJGFiskeRAHietalaSK 1989 Rhodococcus equi foal pneumonia: protective effects of immune plasma in experimentally infected foals. Equine Vet J 21 249 255
55. MadiganJEHietalaSMullerN 1991 Protection against naturally acquired Rhodococcus equi pneumonia in foals by administration of hyperimmune plasma. J Reprod Fertil Suppl 44 571 578
56. HinesSAKanalySTByrneBAPalmerGH 1997 Immunity to Rhodococcus equi. Vet Microbiol 56 177 185
57. HurleyJRBeggAP 1995 Failure of hyperimmune plasma to prevent pneumonia caused by Rhodococcus equi in foals. Aust Vet J 72 418 420
58. GiguèreSGaskinJMMillerCBowmanJL 2002 Evaluation of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals. J Am Vet Med Assoc 220 59 63
59. DawsonTRHorohovDWMeijerWGMuscatelloG 2010 Current understanding of the equine immune response to Rhodococcus equi. An immunological review of R. equi pneumonia. Vet Immunol Immunopathol 135 1 11
60. WeinstockDMBrownAE 2002 Rhodococcus equi: an emerging pathogen. Clin Infect Dis 34 1379 1385
61. RodaRHYoungMTimponeJRosenJ 2009 Rhodococcus equi pulmonary-central nervous system syndrome: brain abscess in a patient on high-dose steroids—a case report and review of the literature. Diagn Microbiol Infect Dis 63 96 99
62. KedlayaIIngMBWongSS 2001 Rhodococcus equi infections in immunocompetent hosts: case report and review. Clin Infect Dis 32 e39 e46
63. De la Pena-MoctezumaAPrescottJF 1995 Association with HeLa cells by Rhodococcus equi with and without the virulence plasmid. Vet Microbiol 46 383 392
64. BenoitSBenachourATaoujiSAuffrayYHartkeA 2001 Induction of vap genes encoded by the virulence plasmid of Rhodococcus equi during acid tolerance response. Res Microbiol 152 439 449
65. VishniacWSanterM 1957 The thiobacilli. Bacteriol Rev 21 195 213
66. SundstromCNilssonK 1976 Establishment and characterization of a human histiocytic lymphoma cell line (U937). Int J Cancer 17 565 577
67. VerbekeGMolenberghsG 2000 Linear mixed models for longitudinal data. New York Springer-Verlag
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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