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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

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