Exoerythrocytic Parasites Secrete a Cysteine Protease Inhibitor Involved in Sporozoite Invasion and Capable of Blocking Cell Death of Host Hepatocytes
Plasmodium parasites must control cysteine protease activity that is critical for hepatocyte invasion by sporozoites, liver stage development, host cell survival and merozoite liberation. Here we show that exoerythrocytic P. berghei parasites express a potent cysteine protease inhibitor (PbICP, P. berghei inhibitor of cysteine proteases). We provide evidence that it has an important function in sporozoite invasion and is capable of blocking hepatocyte cell death. Pre-incubation with specific anti-PbICP antiserum significantly decreased the ability of sporozoites to infect hepatocytes and expression of PbICP in mammalian cells protects them against peroxide- and camptothecin-induced cell death. PbICP is secreted by sporozoites prior to and after hepatocyte invasion, localizes to the parasitophorous vacuole as well as to the parasite cytoplasm in the schizont stage and is released into the host cell cytoplasm at the end of the liver stage. Like its homolog falstatin/PfICP in P. falciparum, PbICP consists of a classical N-terminal signal peptide, a long N-terminal extension region and a chagasin-like C-terminal domain. In exoerythrocytic parasites, PbICP is posttranslationally processed, leading to liberation of the C-terminal chagasin-like domain. Biochemical analysis has revealed that both full-length PbICP and the truncated C-terminal domain are very potent inhibitors of cathepsin L-like host and parasite cysteine proteases. The results presented in this study suggest that the inhibitor plays an important role in sporozoite invasion of host cells and in parasite survival during liver stage development by inhibiting host cell proteases involved in programmed cell death.
Vyšlo v časopise:
Exoerythrocytic Parasites Secrete a Cysteine Protease Inhibitor Involved in Sporozoite Invasion and Capable of Blocking Cell Death of Host Hepatocytes. PLoS Pathog 6(3): e32767. doi:10.1371/journal.ppat.1000825
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1000825
Souhrn
Plasmodium parasites must control cysteine protease activity that is critical for hepatocyte invasion by sporozoites, liver stage development, host cell survival and merozoite liberation. Here we show that exoerythrocytic P. berghei parasites express a potent cysteine protease inhibitor (PbICP, P. berghei inhibitor of cysteine proteases). We provide evidence that it has an important function in sporozoite invasion and is capable of blocking hepatocyte cell death. Pre-incubation with specific anti-PbICP antiserum significantly decreased the ability of sporozoites to infect hepatocytes and expression of PbICP in mammalian cells protects them against peroxide- and camptothecin-induced cell death. PbICP is secreted by sporozoites prior to and after hepatocyte invasion, localizes to the parasitophorous vacuole as well as to the parasite cytoplasm in the schizont stage and is released into the host cell cytoplasm at the end of the liver stage. Like its homolog falstatin/PfICP in P. falciparum, PbICP consists of a classical N-terminal signal peptide, a long N-terminal extension region and a chagasin-like C-terminal domain. In exoerythrocytic parasites, PbICP is posttranslationally processed, leading to liberation of the C-terminal chagasin-like domain. Biochemical analysis has revealed that both full-length PbICP and the truncated C-terminal domain are very potent inhibitors of cathepsin L-like host and parasite cysteine proteases. The results presented in this study suggest that the inhibitor plays an important role in sporozoite invasion of host cells and in parasite survival during liver stage development by inhibiting host cell proteases involved in programmed cell death.
Zdroje
1. SidjanskiS
VanderbergJP
1997 Delayed migration of Plasmodium sporozoites from the mosquito bite site to the blood. Am J Trop Med Hyg 57 426 429
2. MatsuokaH
YoshidaS
HiraiM
IshiiA
2002 A rodent malaria, Plasmodium berghei, is experimentally transmitted to mice by merely probing of infective mosquito, Anopheles stephensi. Parasitol Int 51 17 23
3. VanderbergJP
FrevertU
2004 Intravital microscopy demonstrating antibody-mediated immobilisation of Plasmodium berghei sporozoites injected into skin by mosquitoes. Int J Parasitol 34 991 996
4. MedicaDL
SinnisP
2005 Quantitative dynamics of Plasmodium yoelii sporozoite transmission by infected anopheline mosquitoes. Infect Immun 73 4363 4369
5. AminoR
ThibergeS
MartinB
CelliS
ShorteS
2006 Quantitative imaging of Plasmodium transmission from mosquito to mammal. Nat Med 12 220 224
6. JinY
KebaierC
VanderbergJ
2007 Direct microscopic quantification of dynamics of Plasmodium berghei sporozoite transmission from mosquitoes to mice. Infect Immun 75 5532 5539
7. AminoR
ThibergeS
BlazquezS
BaldacciP
RenaudO
2007 Imaging malaria sporozoites in the dermis of the mammalian host. Nat Protoc 2 1705 1712
8. MotaMM
PradelG
VanderbergJP
HafallaJC
FrevertU
2001 Migration of Plasmodium sporozoites through cells before infection. Science 291 141 144
9. PradelG
FrevertU
2001 Malaria sporozoites actively enter and pass through rat Kupffer cells prior to hepatocyte invasion. Hepatology 33 1154 1165
10. IshinoT
YanoK
ChinzeiY
YudaM
2004 Cell-passage activity is required for the malarial parasite to cross the liver sinusoidal cell layer. PLoS Biol 2 e4 doi:10.1371/journal.pbio.0020004
11. BaerK
RooseveltM
ClarksonABJr
van RooijenN
SchniederT
2007 Kupffer cells are obligatory for Plasmodium yoelii sporozoite infection of the liver. Cell Microbiol 9 397 412
12. AminoR
GiovanniniD
ThibergeS
GueirardP
BoissonB
2008 Host cell traversal is important for progression of the malaria parasite through the dermis to the liver. Cell Host Microbe 3 88 96
13. PrudencioM
RodriguezA
MotaMM
2006 The silent path to thousands of merozoites: the Plasmodium liver stage. Nat Rev Microbiol 4 849 856
14. LeiriaoP
AlbuquerqueSS
CorsoS
van GemertGJ
SauerweinRW
2005 HGF/MET signalling protects Plasmodium-infected host cells from apoptosis. Cell Microbiol 7 603 609
15. van de SandC
HorstmannS
SchmidtA
SturmA
BolteS
2005 The liver stage of Plasmodium berghei inhibits host cell apoptosis. Mol Microbiol 58 731 742
16. SturmA
AminoR
van de SandC
RegenT
RetzlaffS
2006 Manipulation of host hepatocytes by the malaria parasite for delivery into liver sinusoids. Science 313 1287 1290
17. BaerK
KlotzC
KappeSH
SchniederT
FrevertU
2007 Release of hepatic Plasmodium yoelii merozoites into the pulmonary microvasculature. PLoS Pathog 3 e171 doi:10.1371/journal.ppat.0030171
18. RosenthalPJ
2004 Cysteine proteases of malaria parasites. Int J Parasitol 34 1489 1499
19. AlyAS
MatuschewskiK
2005 A malarial cysteine protease is necessary for Plasmodium sporozoite egress from oocysts. J Exp Med 202 225 230
20. CoppiA
Pinzon-OrtizC
HutterC
SinnisP
2005 The Plasmodium circumsporozoite protein is proteolytically processed during cell invasion. J Exp Med 201 27 33
21. CoppiA
TewariR
BishopJR
BennettBL
LawrenceR
2007 Heparan sulfate proteoglycans provide a signal to Plasmodium sporozoites to stop migrating and productively invade host cells. Cell Host Microbe 2 316 327
22. HartmannS
KyewskiB
SonnenburgB
LuciusR
1997 A filarial cysteine protease inhibitor down-regulates T cell proliferation and enhances interleukin-10 production. Eur J Immunol 27 2253 2260
23. ManouryB
GregoryWF
MaizelsRM
WattsC
2001 Bm-CPI-2, a cystatin homolog secreted by the filarial parasite Brugia malayi, inhibits class II MHC-restricted antigen processing. Curr Biol 11 447 451
24. HartmannS
LuciusR
2003 Modulation of host immune responses by nematode cystatins. Int J Parasitol 33 1291 1302
25. SchierackP
LuciusR
SonnenburgB
SchillingK
HartmannS
2003 Parasite-specific immunomodulatory functions of filarial cystatin. Infect Immun 71 2422 2429
26. MurrayJ
ManouryB
BalicA
WattsC
MaizelsRM
2005 Bm-CPI-2, a cystatin from Brugia malayi nematode parasites, differs from Caenorhabditis elegans cystatins in a specific site mediating inhibition of the antigen-processing enzyme AEP. Mol Biochem Parasitol 139 197 203
27. KotsyfakisM
Sa-NunesA
FrancischettiIM
MatherTN
AndersenJF
2006 Antiinflammatory and immunosuppressive activity of sialostatin L, a salivary cystatin from the tick Ixodes scapularis. J Biol Chem 281 26298 26307
28. SchnoellerC
RauschS
PillaiS
AvagyanA
WittigBM
2008 A helminth immunomodulator reduces allergic and inflammatory responses by induction of IL-10-producing macrophages. J Immunol 180 4265 4272
29. Sa-NunesA
BaficaA
AntonelliLR
ChoiEY
FrancischettiIM
2009 The immunomodulatory action of sialostatin L on dendritic cells reveals its potential to interfere with autoimmunity. J Immunol 182 7422 7429
30. HackerG
HawkinsCJ
SmithKG
VauxDL
1996 Effects of viral inhibitors of apoptosis in models of mammalian cell death. Behring Inst Mitt 118 126
31. ZhouQ
SalvesenGS
2000 Viral caspase inhibitors CrmA and p35. Methods Enzymol 322 143 154
32. BlottEJ
GriffithsGM
2002 Secretory lysosomes. Nat Rev Mol Cell Biol 3 122 131
33. LuzioJP
PryorPR
BrightNA
2007 Lysosomes: fusion and function. Nat Rev Mol Cell Biol 8 622 632
34. Jose CazzuloJ
StokaV
TurkV
2001 The major cysteine proteinase of Trypanosoma cruzi: a valid target for chemotherapy of Chagas disease. Curr Pharm Des 7 1143 1156
35. MonteiroAC
AbrahamsonM
LimaAP
Vannier-SantosMA
ScharfsteinJ
2001 Identification, characterization and localization of chagasin, a tight-binding cysteine protease inhibitor in Trypanosoma cruzi. J Cell Sci 114 3933 3942
36. RigdenDJ
MosolovVV
GalperinMY
2002 Sequence conservation in the chagasin family suggests a common trend in cysteine proteinase binding by unrelated protein inhibitors. Protein Sci 11 1971 1977
37. SandersonSJ
WestropGD
ScharfsteinJ
MottramJC
CoombsGH
2003 Functional conservation of a natural cysteine peptidase inhibitor in protozoan and bacterial pathogens. FEBS Lett 542 12 16
38. BesteiroS
CoombsGH
MottramJC
2004 A potential role for ICP, a Leishmanial inhibitor of cysteine peptidases, in the interaction between host and parasite. Mol Microbiol 54 1224 1236
39. RiekenbergS
WitjesB
SaricM
BruchhausI
ScholzeH
2005 Identification of EhICP1, a chagasin-like cysteine protease inhibitor of Entamoeba histolytica. FEBS Lett 579 1573 1578
40. PandeyKC
SinghN
Arastu-KapurS
BogyoM
RosenthalPJ
2006 Falstatin, a cysteine protease inhibitor of Plasmodium falciparum, facilitates erythrocyte invasion. PLoS Pathog 2 e117 doi:10.1371/journal.ppat.0020117
41. SaricM
VahrmannA
BruchhausI
Bakker-GrunwaldT
ScholzeH
2006 The second cysteine protease inhibitor, EhICP2, has a different localization in trophozoites of Entamoeba histolytica than EhICP1. Parasitol Res 100 171 174
42. SatoD
Nakada-TsukuiK
OkadaM
NozakiT
2006 Two cysteine protease inhibitors, EhICP1 and 2, localized in distinct compartments, negatively regulate secretion in Entamoeba histolytica. FEBS Lett 580 5306 5312
43. HuangR
QueX
HirataK
BrinenLS
LeeJH
2009 The cathepsin L of Toxoplasma gondii (TgCPL) and its endogenous macromolecular inhibitor, toxostatin. Mol Biochem Parasitol 164 86 94
44. SantosCC
ScharfsteinJ
LimaAP
2006 Role of chagasin-like inhibitors as endogenous regulators of cysteine proteases in parasitic protozoa. Parasitol Res 99 323 324
45. SantosCC
Sant'annaC
TerresA
Cunha-e-SilvaNL
ScharfsteinJ
2005 Chagasin, the endogenous cysteine-protease inhibitor of Trypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells. J Cell Sci 118 901 915
46. SantosCC
CoombsGH
LimaAP
MottramJC
2007 Role of the Trypanosoma brucei natural cysteine peptidase inhibitor ICP in differentiation and virulence. Mol Microbiol 66 991 1002
47. ClosJ
BrandauS
1994 pJC20 and pJC40–two high-copy-number vectors for T7 RNA polymerase-dependent expression of recombinant genes in Escherichia coli. Protein Expr Purif 5 133 137
48. SchluterA
WiesgiglM
HoyerC
FleischerS
KlaholzL
2000 Expression and subcellular localization of cpn60 protein family members in Leishmania donovani. Biochim Biophys Acta 1491 65 74
49. TurkB
DolencI
TurkV
BiethJG
1993 Kinetics of the pH-induced inactivation of human cathepsin L. Biochemistry 32 375 380
50. HoggT
NagarajanK
HerzbergS
ChenL
ShenX
2006 Structural and functional characterization of Falcipain-2, a hemoglobinase from the malarial parasite Plasmodium falciparum. J Biol Chem 281 25425 25437
51. GraeweS
RetzlaffS
StruckN
JanseCJ
HeusslerVT
2009 Going live: A comparative analysis of the suitability of the RFP derivatives RedStar, mCherry and tdTomato for intravital and in vitro live imaging of Plasmodium parasites. Biotechnol J
52. JanseCJ
RamesarJ
WatersAP
2006 High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium berghei. Nat Protoc 1 346 356
53. KumarKA
OliveiraGA
EdelmanR
NardinE
NussenzweigV
2004 Quantitative Plasmodium sporozoite neutralization assay (TSNA). J Immunol Methods 292 157 164
54. RigdenDJ
MonteiroAC
Grossi de SaMF
2001 The protease inhibitor chagasin of Trypanosoma cruzi adopts an immunoglobulin-type fold and may have arisen by horizontal gene transfer. FEBS Lett 504 41 44
55. SmithBO
PickenNC
WestropGD
BromekK
MottramJC
2006 The structure of Leishmania mexicana ICP provides evidence for convergent evolution of cysteine peptidase inhibitors. J Biol Chem 281 5821 5828
56. SalmonD
do Aido-MachadoR
DiehlA
LeidertM
SchmetzerO
2006 Solution structure and backbone dynamics of the Trypanosoma cruzi cysteine protease inhibitor chagasin. J Mol Biol 357 1511 1521
57. Figueiredo da SilvaAA
de Carvalho VieiraL
KriegerMA
GoldenbergS
ZanchinNI
2007 Crystal structure of chagasin, the endogenous cysteine-protease inhibitor from Trypanosoma cruzi. J Struct Biol 157 416 423
58. RedzyniaI
LjunggrenA
AbrahamsonM
MortJS
KrupaJC
2008 Displacement of the occluding loop by the parasite protein, chagasin, results in efficient inhibition of human cathepsin B. J Biol Chem 283 22815 22825
59. MotaMM
HafallaJC
RodriguezA
2002 Migration through host cells activates Plasmodium sporozoites for infection. Nat Med 8 1318 1322
60. OnoT
Cabrita-SantosL
LeitaoR
BettiolE
PurcellLA
2008 Adenylyl cyclase alpha and cAMP signaling mediate Plasmodium sporozoite apical regulated exocytosis and hepatocyte infection. PLoS Pathog 4 e1000008 doi:10.1371/journal.ppat.1000008
61. StewartMJ
VanderbergJP
1991 Malaria sporozoites release circumsporozoite protein from their apical end and translocate it along their surface. J Protozool 38 411 421
62. BhanotP
FrevertU
NussenzweigV
PerssonC
2003 Defective sorting of the thrombospondin-related anonymous protein (TRAP) inhibits Plasmodium infectivity. Mol Biochem Parasitol 126 263 273
63. GanttS
PerssonC
RoseK
BirkettAJ
AbagyanR
2000 Antibodies against thrombospondin-related anonymous protein do not inhibit Plasmodium sporozoite infectivity in vivo. Infect Immun 68 3667 3673
64. RobsonKJ
FrevertU
ReckmannI
CowanG
BeierJ
1995 Thrombospondin-related adhesive protein (TRAP) of Plasmodium falciparum: expression during sporozoite ontogeny and binding to human hepatocytes. Embo J 14 3883 3894
65. SultanAA
ThathyV
FrevertU
RobsonKJ
CrisantiA
1997 TRAP is necessary for gliding motility and infectivity of plasmodium sporozoites. Cell 90 511 522
66. TreeckM
StruckNS
HaaseS
LangerC
HerrmannS
2006 A conserved region in the EBL proteins is implicated in microneme targeting of the malaria parasite Plasmodium falciparum. J Biol Chem 281 31995 32003
67. ReissM
ViebigN
BrechtS
FourmauxMN
SoeteM
2001 Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii. J Cell Biol 152 563 578
68. LaCrueAN
SivaguruM
WalterMF
FidockDA
JamesAA
2006 A ubiquitous Plasmodium protein displays a unique surface labeling pattern in sporozoites. Mol Biochem Parasitol 148 199 209
69. RawlingsND
MortonFR
BarrettAJ
2006 MEROPS: the peptidase database. Nucleic Acids Res 34 D270 272
70. SilvermanGA
BirdPI
CarrellRW
ChurchFC
CoughlinPB
2001 The serpins are an expanding superfamily of structurally similar but functionally diverse proteins. Evolution, mechanism of inhibition, novel functions, and a revised nomenclature. J Biol Chem 276 33293 33296
71. MusilD
ZucicD
TurkD
EnghRA
MayrI
1991 The refined 2.15 A X-ray crystal structure of human liver cathepsin B: the structural basis for its specificity. Embo J 10 2321 2330
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2010 Číslo 3
- 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
- Kaposi's Sarcoma-Associated Herpesvirus ORF57 Protein Binds and Protects a Nuclear Noncoding RNA from Cellular RNA Decay Pathways
- Endocytosis of the Anthrax Toxin Is Mediated by Clathrin, Actin and Unconventional Adaptors
- Perforin and IL-2 Upregulation Define Qualitative Differences among Highly Functional Virus-Specific Human CD8 T Cells
- Inhibition of Macrophage Migration Inhibitory Factor Ameliorates Ocular -Induced Keratitis