Suppression of mRNAs Encoding Tegument Tetraspanins from Results in Impaired Tegument Turnover
Schistosomes express a family of integral membrane proteins, called tetraspanins (TSPs), in the outer surface membranes of the tegument. Two of these tetraspanins, Sm-TSP-1 and Sm-TSP-2, confer protection as vaccines in mice, and individuals who are naturally resistant to S. mansoni infection mount a strong IgG response to Sm-TSP-2. To determine their functions in the tegument of S. mansoni we used RNA interference to silence expression of Sm-tsp-1 and Sm-tsp-2 mRNAs. Soaking of parasites in Sm-tsp dsRNAs resulted in 61% (p = 0.009) and 74% (p = 0.009) reductions in Sm-tsp-1 and Sm-tsp-2 transcription levels, respectively, in adult worms, and 67%–75% (p = 0.011) and 69%–89% (p = 0.004) reductions in Sm-tsp-1 and Sm-tsp-2 transcription levels, respectively, in schistosomula compared to worms treated with irrelevant control (luciferase) dsRNA. Ultrastructural morphology of adult worms treated in vitro with Sm-tsp-2 dsRNA displayed a distinctly vacuolated and thinner tegument compared with controls. Schistosomula exposed in vitro to Sm-tsp-2 dsRNA had a significantly thinner and more vacuolated tegument, and morphology consistent with a failure of tegumentary invaginations to close. Injection of mice with schistosomula that had been electroporated with Sm-tsp-1 and Sm-tsp-2 dsRNAs resulted in 61% (p = 0.005) and 83% (p = 0.002) reductions in the numbers of parasites recovered from the mesenteries four weeks later when compared to dsRNA-treated controls. These results imply that tetraspanins play important structural roles impacting tegument development, maturation or stability.
Vyšlo v časopise:
Suppression of mRNAs Encoding Tegument Tetraspanins from Results in Impaired Tegument Turnover. PLoS Pathog 6(4): e32767. doi:10.1371/journal.ppat.1000840
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1000840
Souhrn
Schistosomes express a family of integral membrane proteins, called tetraspanins (TSPs), in the outer surface membranes of the tegument. Two of these tetraspanins, Sm-TSP-1 and Sm-TSP-2, confer protection as vaccines in mice, and individuals who are naturally resistant to S. mansoni infection mount a strong IgG response to Sm-TSP-2. To determine their functions in the tegument of S. mansoni we used RNA interference to silence expression of Sm-tsp-1 and Sm-tsp-2 mRNAs. Soaking of parasites in Sm-tsp dsRNAs resulted in 61% (p = 0.009) and 74% (p = 0.009) reductions in Sm-tsp-1 and Sm-tsp-2 transcription levels, respectively, in adult worms, and 67%–75% (p = 0.011) and 69%–89% (p = 0.004) reductions in Sm-tsp-1 and Sm-tsp-2 transcription levels, respectively, in schistosomula compared to worms treated with irrelevant control (luciferase) dsRNA. Ultrastructural morphology of adult worms treated in vitro with Sm-tsp-2 dsRNA displayed a distinctly vacuolated and thinner tegument compared with controls. Schistosomula exposed in vitro to Sm-tsp-2 dsRNA had a significantly thinner and more vacuolated tegument, and morphology consistent with a failure of tegumentary invaginations to close. Injection of mice with schistosomula that had been electroporated with Sm-tsp-1 and Sm-tsp-2 dsRNAs resulted in 61% (p = 0.005) and 83% (p = 0.002) reductions in the numbers of parasites recovered from the mesenteries four weeks later when compared to dsRNA-treated controls. These results imply that tetraspanins play important structural roles impacting tegument development, maturation or stability.
Zdroje
1. FenwickA
RollinsonD
SouthgateV
2006 Implementation of human schistosomiasis control: Challenges and prospects. Adv Parasitol 61 567 622
2. GryseelsB
PolmanK
ClerinxJ
KestensL
2006 Human schistosomiasis. Lancet 368 1106 1118
3. SteinmannP
KeiserJ
BosR
TannerM
UtzingerJ
2006 Schistosomiasis and water resources development: systematic review, meta-analysis, and estimates of people at risk. Lancet Infect Dis 6 411 425
4. MillerP
WilsonRA
1980 Migration of the schistosomula of Schistosoma mansoni from the lungs to the hepatic portal system. Parasitology 80 267 288
5. JonesMK
GobertGN
ZhangL
SunderlandP
McManusDP
2004 The cytoskeleton and motor proteins of human schistosomes and their roles in surface maintenance and host-parasite interactions. Bioessays 26 752 765
6. SmithersSR
McLarenDJ
Ramalho-PintoFJ
1977 Immunity to schistosomes: the target. Am J Trop Med Hyg 26 11 19
7. SmithersSR
TerryRJ
1976 The immunology of schistosomiasis. Adv Parasitol 14 399 422
8. LoukasA
TranM
PearsonMS
2007 Schistosome membrane proteins as vaccines. Int J Parasitol 37 257 263
9. Verjovski-AlmeidaS
DeMarcoR
MartinsEA
GuimaraesPE
OjopiEP
2003 Transcriptome analysis of the acoelomate human parasite Schistosoma mansoni. Nat Genet 35 148 157
10. BerrimanM
HaasBJ
LoVerdePT
WilsonRA
DillonGP
2009 The genome of the blood fluke Schistosoma mansoni. Nature 460 352 358
11. BraschiS
CurwenRS
AshtonPD
Verjovski-AlmeidaS
WilsonA
2006 The tegument surface membranes of the human blood parasite Schistosoma mansoni: a proteomic analysis after differential extraction. Proteomics 6 1471 1482
12. GaugitschHW
HoferE
HuberNE
SchnablE
BaumrukerT
1991 A new superfamily of lymphoid and melanoma cell proteins with extensive homology to Schistosoma mansoni antigen Sm23. Eur J Immunol 21 377 383
13. SmythD
McManusDP
SmoutMJ
LahaT
ZhangW
2003 Isolation of cDNAs encoding secreted and transmembrane proteins from Schistosoma mansoni by a signal sequence trap method. Infect Immun 71 2548 2554
14. BraschiS
BorgesWC
WilsonRA
2006 Proteomic analysis of the schistosome tegument and its surface membranes. Mem Inst Oswaldo Cruz 101 Suppl 1 205 212
15. KovalenkoOV
MetcalfDG
DeGradoWF
HemlerME
2005 Structural organization and interactions of transmembrane domains in tetraspanin proteins. BMC Struct Biol 5 11
16. HemlerME
2001 Specific tetraspanin functions. J Cell Biol 155 1103 1107
17. StippCS
KolesnikovaTV
HemlerME
2003 Functional domains in tetraspanin proteins. Trends Biochem Sci 28 106 112
18. BoucheixC
DucGH
JasminC
RubinsteinE
2001 Tetraspanins and malignancy. Expert Rev Mol Med 2001 1 17
19. HemlerME
2003 Tetraspanin proteins mediate cellular penetration, invasion, and fusion events and define a novel type of membrane microdomain. Annu Rev Cell Dev Biol 19 397 422
20. WrightMD
MoseleyGW
van SprielAB
2004 Tetraspanin microdomains in immune cell signalling and malignant disease. Tissue Antigens 64 533 542
21. HigginbottomA
TakahashiY
BollingL
CoonrodSA
WhiteJM
2003 Structural requirements for the inhibitory action of the CD9 large extracellular domain in sperm/oocyte binding and fusion. Biochem Biophys Res Commun 311 208 214
22. LazoPA
2007 Functional implications of tetraspanin proteins in cancer biology. Cancer Sci 98 1666 1677
23. RubinsteinE
ZiyyatA
PrenantM
WrobelE
WolfJP
2006 Reduced fertility of female mice lacking CD81. Dev Biol 290 351 358
24. Barraud-LangeV
Naud-BarriantN
BomselM
WolfJP
ZiyyatA
2007 Transfer of oocyte membrane fragments to fertilizing spermatozoa. FASEB J 21 3446 3449
25. Gordon-AlonsoM
Yanez-MoM
BarreiroO
AlvarezS
Munoz-FernandezMA
2006 Tetraspanins CD9 and CD81 modulate HIV-1-induced membrane fusion. J Immunol 177 5129 5137
26. BartoschB
VitelliA
GranierC
GoujonC
DubuissonJ
2003 Cell entry of hepatitis C virus requires a set of co-receptors that include the CD81 tetraspanin and the SR-B1 scavenger receptor. J Biol Chem 278 41624 41630
27. YalaouiS
ZougbedeS
CharrinS
SilvieO
ArduiseC
2008 Hepatocyte permissiveness to Plasmodium infection is conveyed by a short and structurally conserved region of the CD81 large extracellular domain. PLoS Pathog 4 e1000010 doi:10.1371/journal.ppat.1000010
28. TranMH
PearsonMS
BethonyJM
SmythDJ
JonesMK
2006 Tetraspanins on the surface of Schistosoma mansoni are protective antigens against schistosomiasis. Nature Med 12 835 840
29. van BalkomBW
van GestelRA
BrouwersJF
KrijgsveldJ
TielensAG
2005 Mass spectrometric analysis of the Schistosoma mansoni tegumental sub-proteome. J Proteome Res 4 958 966
30. BraschiS
WilsonRA
2006 Proteins exposed at the adult schistosome surface revealed by biotinylation. Mol Cell Proteomics 5 347 356
31. CorrentiJM
PearceEJ
2004 Transgene expression in Schistosoma mansoni: introduction of RNA into schistosomula by electroporation. Mol Biochem Parasitol 137 75 79
32. SkellyPJ
Da'daraA
HarnDA
2003 Suppression of cathepsin B expression in Schistosoma mansoni by RNA interference. Int J Parasitol 33 363 369
33. FreitasTC
JungE
PearceEJ
2007 TGF-beta signaling controls embryo development in the parasitic flatworm Schistosoma mansoni. PLoS Pathog 3 e52 doi:10.1371/journal.ppat.0030052
34. BoyleJP
WuXJ
ShoemakerCB
YoshinoTP
2003 Using RNA interference to manipulate endogenous gene expression in Schistosoma mansoni sporocysts. Mol Biochem Parasitol 128 205 215
35. HockleyDJ
McLarenDJ
1973 Schistosoma mansoni: changes in the outer membrane of the tegument during development from cercaria to adult worm. Int J Parasitol 3 13 25
36. BaschPF
1981 Cultivation of Schistosoma mansoni in vitro. I. Establishment of cultures from cercariae and development until pairing. J Parasitol 67 179 185
37. SchmidtJ
BodorO
GohrL
KunzW
1996 Paramyosin isoforms of Schistosoma mansoni are phosphorylated and localized in a large variety of muscle types. Parasitology 112 459 467
38. CousinCE
StirewaltMA
DorseyCH
WatsonLP
1986 Schistosoma mansoni: comparative development of schistosomules produced by artificial techniques. J Parasitol 72 606 609
39. Da'daraAA
LiYS
XiongT
ZhouJ
WilliamsGM
2008 DNA-based vaccines protect against zoonotic schistosomiasis in water buffalo. Vaccine 26 3617 3625
40. NdegwaD
Krautz-PetersonG
SkellyPJ
2007 Protocols for gene silencing in schistosomes. Exp Parasitol 117 284 291
41. PearceEJ
FreitasTC
2008 Reverse genetics and the study of the immune response to schistosomes. Parasite Immunol 30 215 221
42. CorrentiJM
BrindleyPJ
PearceEJ
2005 Long-term suppression of cathepsin B levels by RNA interference retards schistosome growth. Mol Biochem Parasitol 143 209 215
43. MoralesME
RinaldiG
GobertGN
KinesKJ
TortJF
2008 RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade. Mol Biochem Parasitol 157 160 168
44. FaghiriZ
SkellyPJ
2009 The role of tegumental aquaporin from the human parasitic worm, Schistosoma mansoni, in osmoregulation and drug uptake. FASEB J 23 2780 2789
45. HemlerME
2008 Targeting of tetraspanin proteins - potential benefits and strategies. Nature Rev Drug Discov 7 747 758
46. MoribeH
YochemJ
YamadaH
TabuseY
FujimotoT
2004 Tetraspanin protein (TSP-15) is required for epidermal integrity in Caenorhabditis elegans. J Cell Sci 117 5209 5220
47. YamadaM
SumidaY
FujibayashiA
FukaguchiK
SanzenN
2008 The tetraspanin CD151 regulates cell morphology and intracellular signaling on laminin-511. FEBS J 275 3335 3351
48. SzollosiJ
HorejsiV
BeneL
AngelisovaP
DamjanovichS
1996 Supramolecular complexes of MHC class I, MHC class II, CD20, and tetraspan molecules (CD53, CD81, and CD82) at the surface of a B cell line JY. J Immunol 157 2939 2946
49. KropshoferH
SpindeldreherS
RohnTA
PlataniaN
GrygarC
2002 Tetraspan microdomains distinct from lipid rafts enrich select peptide-MHC class II complexes. Nature Immunol 3 61 68
50. ShengKC
van SprielAB
GartlanKH
SofiM
ApostolopoulosV
2009 Tetraspanins CD37 and CD151 differentially regulate Ag presentation and T-cell co-stimulation by DC. Eur J Immunol 39 50 55
51. SherA
HallBF
VadasMA
1978 Acquisition of murine major histocompatibility complex gene products by schistosomula of Schistosoma mansoni. J Exp Med 148 46 57
52. Le NaourF
AndreM
GrecoC
BillardM
SordatB
2006 Profiling of the tetraspanin web of human colon cancer cells. Mol Cell Proteomics 5 845 857
53. LiuL
HeB
LiuWM
ZhouD
CoxJV
2007 Tetraspanin CD151 promotes cell migration by regulating integrin trafficking. J Biol Chem 282 31631 31642
54. YangXH
RichardsonAL
Torres-ArzayusMI
ZhouP
SharmaC
2008 CD151 accelerates breast cancer by regulating alpha 6 integrin function, signaling, and molecular organization. Cancer Res 68 3204 3213
55. DeisslerH
KuhnEM
LangGE
2007 Tetraspanin CD9 is involved in the migration of retinal microvascular endothelial cells. Int J Mol Med 20 643 652
56. JamesER
TaylorMG
1976 Transformation of cercariae to schistosomula: a quantitative comparison of transformation techniques and of infectivity by different injection routes of the organisms produced. J Helminthol 50 223 233
57. SalafskyB
FuscoAC
WhitleyK
NowickiD
EllenbergerB
1988 Schistosoma mansoni: analysis of cercarial transformation methods. Exp Parasitol 67 116 127
58. LazdinsJK
SteinMJ
DavidJR
SherA
1982 Schistosoma mansoni: rapid isolation and purification of schistosomula of different developmental stages by centrifugation on discontinuous density gradients of Percoll. Exp Parasitol 53 39 44
59. LivakKJ
SchmittgenTD
2001 Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25 402 408
60. PearceEJ
JamesSL
DaltonJ
BarrallA
RamosC
1986 Immunochemical characterization and purification of Sm-97, a Schistosoma mansoni antigen monospecifically recognized by antibodies from mice protectively immunized with a nonliving vaccine. J Immunol 137 3593 3600
61. GriffithsG
1993 Fine Structure Immunocytochemistry Berlin Springer 459
62. BartleyPB
RammGA
JonesMK
RuddellRG
LiY
McManusDP
2006 A contributory role for activated hepatic stellate cells in the dynamics of Schistosoma japonicum egg-induced fibrosis. Int J Parasitol 36 993 1001
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2010 Číslo 4
- 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
- The Effect of Vaccination on the Evolution and Population Dynamics of Avian Paramyxovirus-1
- Reconstitution of SARS-Coronavirus mRNA Cap Methylation
- Deficiencies in Jasmonate-Mediated Plant Defense Reveal Quantitative Variation in Pathogenesis
- A Timescale for Evolution, Population Expansion, and Spatial Spread of an Emerging Clone of Methicillin-Resistant