Molecular Basis of Increased Serum Resistance among Pulmonary Isolates of Non-typeable
Non-typeable Haemophilus influenzae (NTHi), a common commensal of the human pharynx, is also an opportunistic pathogen if it becomes established in the lower respiratory tract (LRT). In comparison to colonizing isolates from the upper airway, LRT isolates, especially those associated with exacerbations of chronic obstructive pulmonary disease, have increased resistance to the complement- and antibody-dependent, bactericidal effect of serum. To define the molecular basis of this resistance, mutants constructed in a serum resistant strain using the mariner transposon were screened for loss of survival in normal human serum. The loci required for serum resistance contribute to the structure of the exposed surface of the bacterial outer membrane. These included loci involved in biosynthesis of the oligosaccharide component of lipooligosaccharide (LOS), and vacJ, which functions with an ABC transporter encoded by yrb genes in retrograde trafficking of phospholipids from the outer to inner leaflet of the cell envelope. Mutations in vacJ and yrb genes reduced the stability of the outer membrane and were associated with increased cell surface hyrophobicity and phospholipid content. Loss of serum resistance in vacJ and yrb mutants correlated with increased binding of natural immunoglobulin M in serum as well as anti-oligosaccharide mAbs. Expression of vacJ and the yrb genes was positively correlated with serum resistance among clinical isolates. Our findings suggest that NTHi adapts to inflammation encountered during infection of the LRT by modulation of its outer leaflet through increased expression of vacJ and yrb genes to minimize recognition by bactericidal anti-oligosaccharide antibodies.
Vyšlo v časopise:
Molecular Basis of Increased Serum Resistance among Pulmonary Isolates of Non-typeable. PLoS Pathog 7(1): e32767. doi:10.1371/journal.ppat.1001247
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1001247
Souhrn
Non-typeable Haemophilus influenzae (NTHi), a common commensal of the human pharynx, is also an opportunistic pathogen if it becomes established in the lower respiratory tract (LRT). In comparison to colonizing isolates from the upper airway, LRT isolates, especially those associated with exacerbations of chronic obstructive pulmonary disease, have increased resistance to the complement- and antibody-dependent, bactericidal effect of serum. To define the molecular basis of this resistance, mutants constructed in a serum resistant strain using the mariner transposon were screened for loss of survival in normal human serum. The loci required for serum resistance contribute to the structure of the exposed surface of the bacterial outer membrane. These included loci involved in biosynthesis of the oligosaccharide component of lipooligosaccharide (LOS), and vacJ, which functions with an ABC transporter encoded by yrb genes in retrograde trafficking of phospholipids from the outer to inner leaflet of the cell envelope. Mutations in vacJ and yrb genes reduced the stability of the outer membrane and were associated with increased cell surface hyrophobicity and phospholipid content. Loss of serum resistance in vacJ and yrb mutants correlated with increased binding of natural immunoglobulin M in serum as well as anti-oligosaccharide mAbs. Expression of vacJ and the yrb genes was positively correlated with serum resistance among clinical isolates. Our findings suggest that NTHi adapts to inflammation encountered during infection of the LRT by modulation of its outer leaflet through increased expression of vacJ and yrb genes to minimize recognition by bactericidal anti-oligosaccharide antibodies.
Zdroje
1. MukundanD
EcevitZ
PatelM
MarrsC
GilsdorfJ
2007 Pharyngeal colonization dynamics of Haemophilus influenzae and Haemophilus haemolyticus in healthy adult carriers. J Clin Microbiol 45 3207 3217
2. MurphyT
2003 Respiratory infections caused by non-typeable Haemophilus influenzae. Curr Opin Infect Dis 16 129 134
3. TurkDC
1984 The pathogenicity of Haemophilus influenzae. J Med Microbiol 18 1 16
4. SethiS
EvansN
GrantBJ
MurphyTF
2002 New strains of bacteria and exacerbations of chronic obstructive pulmonary disease. N Engl J Med 347 465 471
5. BandiV
ApicellaMA
MasonE
MurphyTF
SiddiqiA
2001 Nontypeable Haemophilus influenzae in the lower respiratory tract of patients with chronic bronchitis. Am J Respir Crit Care Med 164 2114 2119
6. PauwelsR
BuistA
CalverleyP
JenkinsC
HurdS
2001 Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease. NHLBI/WHO Global Initiative for Chronic Obstructive Lung Disease (GOLD) Workshop summary. Am J Respir Crit Care Med 163 1256 1276
7. MurrayC
LopezA
1997 Alternative projections of mortality and disability by cause 1990-2020: global burden of disease study. Lancet 349 1498 1504
8. NikaidoH
2003 Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 67 593 656
9. JiaW
El ZoeibyA
PetruzzielloTN
JayabalasinghamB
SeyedirashtiS
2004 Lipid trafficking controls endotoxin acylation in outer membranes of Escherichia coli. J Biol Chem 279 44966 44975
10. KamioY
NikaidoH
1976 Outer membrane of Salmonella typhimurium: accessibility of phospholipid head groups to phospholipase c and cyanogen bromide activated dextran in the external medium. Biochemistry 15 2561 2570
11. SwordsW
JonesP
ApicellaM
2003 The lipo-oligosaccharides of Haemophilus influenzae: an interesting array of characters. J Endotoxin Res 9 131 134
12. SchwedaE
RichardsJ
HoodD
MoxonE
2007 Expression and structural diversity of the lipopolysaccharide of Haemophilus influenzae: implication in virulence. Int J Med Microbiol 297 297 306
13. WeiserJN
PanN
1998 Adaptation of Haemophilus influenzae to acquired and innate humoral immunity based on phase variation of lipopolysaccharide. Mol Microbiol 30 767 775
14. ErwinAL
AllenS
HoDK
BonthuisPJ
JarischJ
2006 Role of lgtC in resistance of nontypeable Haemophilus influenzae strain R2866 to human serum. Infect Immun 74 6226 6235
15. HoDK
RamS
NelsonKL
BonthuisPJ
SmithAL
2007 lgtC expression modulates resistance to C4b deposition on an invasive nontypeable Haemophilus influenzae. J Immunol 178 1002 1012
16. FigueiraM
RamS
GoldsteinR
HoodD
MoxonE
2007 Role of complement in defense of the middle ear revealed by restoring the virulence of nontypeable Haemophilus influenzae siaB mutants. Infect Immun 75 325 333
17. WeiserJN
LoveJM
MoxonER
1989 The molecular mechanism of phase variation of H. influenzae lipopolysaccharide. Cell 59 657 665
18. WilliamsBJ
MorlinG
ValentineN
SmithAL
2001 Serum resistance in an invasive, nontypeable Haemophilus influenzae strain. Infect Immun 69 695 705
19. HoodD
DeadmanM
AllenT
MasoudH
MartinA
1996 Use of the complete genome sequence information of Haemophilus influenzae strain Rd to investigate lipopolysaccharide biosynthesis. Mol Microbiol 22 951 965
20. MalinverniJC
SilhavyTJ
2009 An ABC transport system that maintains lipid asymmetry in the gram-negative outer membrane. Proc Natl Acad Sci U S A 106 8009 8014
21. HaaseE
CampagnariA
SarwarJ
SheroM
WirthM
1991 Strain specific and immunodominant surface epitopes of the P2 porin protein of nontypeable Haemophilus influenzae. Infect Immun 59 1278 1284
22. OchsenbeinAF
FehrT
LutzC
SuterM
BrombacherF
1999 Control of early viral and bacterial distribution and disease by natural antibodies. Science 286 2156 2159
23. FadenH
DuffyL
WilliamsA
KrystofikD
WolfJ
1996 Epidemiology of nasopharyngeal colonization with nontypeable Haemophilus influenzae in the first two years of life. Acta Otolaryngol Suppl 523 128 129
24. OksenhendlerE
GérardL
FieschiC
MalphettesM
MouillotG
2008 Infections in 252 patients with common variable immunodeficiency. Clin Infect Dis 46 1547 1554
25. ErwinA
BrewahY
CouchenourD
BarrenP
BurkeS
2000 Role of lipopolysaccharide phase variation in susceptibility of Haemophilus influenzae to bactericidal immunoglobulin M antibodies in rabbit sera. Infect Immun 68 2804 2807
26. ZolaTA
LysenkoES
WeiserJN
2009 Natural antibody to conserved targets of Haemophilus influenzae limits colonization of the murine nasopharynx. Infect Immun 77 3458 3465
27. ReidR
ProdeusA
KhanW
HsuT
RosenF
1997 Endotoxin shock in antibody-deficient mice: unraveling the role of natural antibody and complement in the clearance of lipopolysaccharide. J Immunol 159 970 975
28. BoesM
ProdeusA
SchmidtT
CarrollM
ChenJ
1998 A critical role of natural immunoglobulin M in immediate defense against systemic bacterial infection. J Exp Med 188 2381 2386
29. SuzukiT
MuraiT
FukudaI
TobeT
YoshikawaM
1994 Identification and characterization of a chromosomal virulence gene, vacJ, required for intercellular spreading of Shigella flexneri. Mol Microbiol 11 31 41
30. NikaidoH
VaaraM
1985 Molecular Basis of Bacterial Outer Menbrane Permeability. Microbiol Rev 49 1 32
31. St SauverJ
MarrsC
FoxmanB
SomselP
MaderaR
2000 Risk factors for otitis media and carriage of multiple strains of Haemophilus influenzae and Streptococcus pneumoniae. Emerg Infect Dis 6 622 630
32. FarjoR
FoxmanB
PatelM
ZhangL
PettigrewM
2004 Diversity and sharing of Haemophilus influenzae strains colonizing healthy children attending day-care centers. Pediatr Infect Dis J 23 41 46
33. McCreaK
XieJ
LaCrossN
PatelM
MukundanM
2008 Relationships of non-typeable Haemophilus influenzae with hemolytic and non-hemolytic Haemophilus haemolyticus. J Clin Microbiol 46 406 416
34. LampeDJ
ChurchillME
RobertsonHM
1996 A purified mariner transposase is sufficient to mediate transposition in vitro. Embo J 15 5470 5479
35. MartinB
PrudhommeM
AlloingG
GranadelC
ClaverysJP
2000 Cross-regulation of competence pheromone production and export in the early control of transformation in Streptococcus pneumoniae. Mol Microbiol 38 867 878
36. AkerleyB
RubinE
CamilliA
LampeD
RobertsonH
1998 Systematic identification of essential genes by in vitro mariner mutagenesis. Proc Natl Acad Sci U S A 95 8927 8932
37. HerriottRM
MeyerEM
VogtM
1970 Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol 101 517 524
38. MatsushitaM
OkadaH
1986 Alternative complement pathway activation by C4b deposited during classical pathway activation. J Immunol 136 2994 2998
39. WeiserJN
ShchepetovM
ChongST
1997 Decoration of lipopolysaccharide with phosphorylcholine: a phase-variable characteristic of Haemophilus influenzae. Infect Immun 65 943 950
40. GuligPA
PatrickCC
HermanstorferL
McCrackenGHJr
HansenEJ
1987 Conservation of epitopes in the oligosaccharide portion of the lipooligosaccharide of Haemophilus influenzae type b. Infect Immun 55 513 520
41. KhemiriA
GallandA
VaudryD
Chan Tchi SongP
VaudryH
2008 Outer-membrane proteomic maps and surface-exposed proteins of Legionella pneumophila using cellular fractionation and fluorescent labelling. Anal Bioanal Chem 390 1861 1871
42. GiufreM
CarattoliA
CardinesR
MastrantonioP
CerquettiM
2008 Variation in expression of HMW1 and HMW2 adhesins in invasive nontypeable Haemophilus influenzae isolates. BMC Microbiol 8 83
43. Martinez de TejadaG
MorivónI
1993 The outer membranes of Brucella spp. are not barriers to hydrophobic permeants. J Bacteriol 175 5273 5275
44. GoldfineH
JohnstonN
BishopD
1982 Ether phospholipid asymmetry in Clostridium butyricum. Biochem Biophys Res Commun 108 1502 1507
45. BlighE
DyerW
1959 A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37 911 917
46. FleischmannRD
AdamsMD
WhiteO
ClaytonRA
KirknessEF
1995 Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269 496 512
47. PhillipsNJ
ApicellaMA
GriffissJM
GibsonBW
1992 Structural characterization of the cell surface lipooligosaccharides from a nontypable strain of Haemophilus influenzae. Biochemistry 31 4515 4526
48. MerrellDS
HavaDL
CamilliA
2002 Identification of novel factors involved in colonization and acid tolerance of Vibrio cholerae. Mol Microbiol 43 1471 1491
49. HavaDL
CamilliA
2002 Large-scale identification of serotype 4 Streptococcus pneumoniae virulence factors. Mol Microbiol 45 1389 1406
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2011 Číslo 1
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- Dual-Use Research and Technological Diffusion: Reconsidering the Bioterrorism Threat Spectrum
- Pathogenesis of the 1918 Pandemic Influenza Virus
- Critical Role of IRF-5 in the Development of T helper 1 responses to infection
- A Cardinal Role for Cathepsin D in Co-Ordinating the Host-Mediated Apoptosis of Macrophages and Killing of Pneumococci