Infections in Cells: Transcriptomic Characterization of a Novel Host-Symbiont Interaction
The endosymbiotic bacterium Wolbachia is being investigated as a potential control agent in several important vector insect species. Recent studies have shown that Wolbachia can protect the insect host against a wide variety of pathogens, resulting in reduced transmission of parasites and viruses. It has been proposed that compromised vector competence of Wolbachia-infected insects is due to up-regulation of the host innate immune system or metabolic competition. Anopheles mosquitoes, which transmit human malaria parasites, have never been found to harbor Wolbachia in nature. While transient somatic infections can be established in Anopheles, no stable artificially-transinfected Anopheles line has been developed despite numerous attempts. However, cultured Anopheles cells can be stably infected with multiple Wolbachia strains such as wAlbB from Aedes albopictus, wRi from Drosophila simulans and wMelPop from Drosophila melanogaster. Infected cell lines provide an amenable system to investigate Wolbachia-Anopheles interactions in the absence of an infected mosquito strain. We used Affymetrix GeneChip microarrays to investigate the effect of wAlbB and wRi infection on the transcriptome of cultured Anopheles Sua5B cells, and for a subset of genes used quantitative PCR to validate results in somatically-infected Anopheles mosquitoes. Wolbachia infection had a dramatic strain-specific effect on gene expression in this cell line, with almost 700 genes in total regulated representing a diverse array of functional classes. Very strikingly, infection resulted in a significant down-regulation of many immune, stress and detoxification-related transcripts. This is in stark contrast to the induction of immune genes observed in other insect hosts. We also identified genes that may be potentially involved in Wolbachia-induced reproductive and pathogenic phenotypes. Somatically-infected mosquitoes had similar responses to cultured cells. The data show that Wolbachia has a profound and unique effect on Anopheles gene expression in cultured cells, and has important implications for mechanistic understanding of Wolbachia-induced phenotypes and potential novel strategies to control malaria.
Vyšlo v časopise:
Infections in Cells: Transcriptomic Characterization of a Novel Host-Symbiont Interaction. PLoS Pathog 7(2): e32767. doi:10.1371/journal.ppat.1001296
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1001296
Souhrn
The endosymbiotic bacterium Wolbachia is being investigated as a potential control agent in several important vector insect species. Recent studies have shown that Wolbachia can protect the insect host against a wide variety of pathogens, resulting in reduced transmission of parasites and viruses. It has been proposed that compromised vector competence of Wolbachia-infected insects is due to up-regulation of the host innate immune system or metabolic competition. Anopheles mosquitoes, which transmit human malaria parasites, have never been found to harbor Wolbachia in nature. While transient somatic infections can be established in Anopheles, no stable artificially-transinfected Anopheles line has been developed despite numerous attempts. However, cultured Anopheles cells can be stably infected with multiple Wolbachia strains such as wAlbB from Aedes albopictus, wRi from Drosophila simulans and wMelPop from Drosophila melanogaster. Infected cell lines provide an amenable system to investigate Wolbachia-Anopheles interactions in the absence of an infected mosquito strain. We used Affymetrix GeneChip microarrays to investigate the effect of wAlbB and wRi infection on the transcriptome of cultured Anopheles Sua5B cells, and for a subset of genes used quantitative PCR to validate results in somatically-infected Anopheles mosquitoes. Wolbachia infection had a dramatic strain-specific effect on gene expression in this cell line, with almost 700 genes in total regulated representing a diverse array of functional classes. Very strikingly, infection resulted in a significant down-regulation of many immune, stress and detoxification-related transcripts. This is in stark contrast to the induction of immune genes observed in other insect hosts. We also identified genes that may be potentially involved in Wolbachia-induced reproductive and pathogenic phenotypes. Somatically-infected mosquitoes had similar responses to cultured cells. The data show that Wolbachia has a profound and unique effect on Anopheles gene expression in cultured cells, and has important implications for mechanistic understanding of Wolbachia-induced phenotypes and potential novel strategies to control malaria.
Zdroje
1. WerrenJH
BaldoL
ClarkME
2008 Wolbachia: master manipulators of invertebrate biology. Nat Rev Microbiol 6 741 751
2. KambrisZ
CookPE
PhucHK
SinkinsSP
2009 Immune activation by life-shortening Wolbachia and reduced filarial competence in mosquitoes. Science 326 134 136
3. MoreiraLA
Iturbe-OrmaetxeI
JefferyJA
LuG
PykeAT
2009 A Wolbachia symbiont in Aedes aegypti limits infection with Dengue, Chikungunya, and Plasmodium. Cell 139 1268 1278
4. HedgesLM
BrownlieJC
O'NeillSL
JohnsonKN
2008 Wolbachia and virus protection in insects. Science 322 702 702
5. TeixeiraL
FerreiraA
AshburnerM
2008 The bacterial symbiont Wolbachia induces resistance to RNA viral infections in Drosophila melanogaster. PloS Biol 6 e2
6. BianG
XuY
LuP
XieY
XiZ
2010 The endosymbiotic bacterium Wolbachia induces resistance to Dengue virus in Aedes aegypti. PLoS Pathog 6 e1000833
7. KambrisZ
BlagboroughAM
PintoSB
BlagroveMSC
GodfrayHCJ
2010 Wolbachia stimulates immune gene expression and inhibits Plasmodium development in Anopheles gambiae. PLoS Pathog 6 e1001143
8. SinkinsSP
O'NeillSL
2000 Wolbachia as a vehicle to modify insect populations.
HandlerAF
JamesAA
Insect transgenesis: methods and applications New York CRC Press 271 287
9. SinkinsSP
CurtisC
O'NeillSL
1997 The potential application of inherited symbiont systems to pest control.
O'NeillSL
HoffmannAA
WerrenJH
Influential passengers Oxford Oxford University Press 155 175
10. KittayapongP
BaisleyKJ
BaimaiV
O'NeillSL
2000 Distribution and diversity of Wolbachia infections in southeast Asian mosquitoes (Diptera: Culicidea). J Med Entomol 37 340 345
11. RasgonJL
ScottTW
2004 An initial survey for Wolbachia (Rickettsiales: Rickettsiaceae) infections in selected California mosquitoes (Diptera: Culicidae). J Med Entomol 41 255 257
12. RicciI
CancriniG
GabrielliS
D'AmelioS
FaviG
2002 Searching for Wolbachia (Rickettsiales: Rickettsiaceae) in mosquitoes (Diptera: Culicidae): large polymerase chain reaction survey and new identifications. J Med Entomol 39 562 567
13. JinC
RenX
RasgonJL
2009 The virulent Wolbachia strain wMelPop efficiently establishes somatic infections in the malaria vector Anopheles gambiae. Appl Environ Microbiol 75 3373 3376
14. RasgonJL
RenX
PetridisM
2006 Can Anopheles gambiae be infected with Wolbachia pipientis? Insights from an in vitro system. Appl Environ Microbiol 72 7718 7722
15. SuhE
MercerD
FuY
DobsonS
2009 Life-shortening Wolbachia infection from Drosophila melanogaster transferred into Aedes albopictus is pathogenic. Appl Environ Microbiol 75 7783 7788
16. XiZ
KhooC
DobsonSL
2006 Interspecific transfer of Wolbachia into the mosquito disease vector Aedes albopictus. Proc R Soc Lond B 273 1317 1322
17. McMenimanCJ
LaneAM
CassBN
FongAWC
SidhuM
2009 Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti. Science 323 141 144
18. XiZ
DeanJ
KhooC
DobsonSL
2005 Generation of artificial Wolbachia infections in Aedes mosquito and manipulation of population with cytoplasmic incompatibilty. Am J Trop Med Hyg 73 349 349
19. XiZ
DeanJ
KhooC
DobsonSL
2005 Generation of a novel Wolbachia infection in Aedes albopictus (Asian tiger mosquito) via embryonic microinjection. Insect Biochem Mol Biol 35 903 910
20. XiZ
KhooC
DobsonSL
2005 Wolbachia establishment and invasion in an Aedes aegypti laboratory population. Science 310 326 328
21. McGrawEA
MerrittDJ
DrollerJN
O'NeillSL
2001 Wolbachia-mediated sperm modification is dependent on the host genotype in Drosophila. Proc R Soc Lond B 268 2565 2570
22. McGrawEA
MerrittDJ
DrollerJN
O'NeillSL
2002 Wolbachia density and virulence attenuation after transfer into a novel host. Proc Natl Acad Sci USA 99 2918 2923
23. BourtzisK
PettigrewMM
O'NeillSL
2000 Wolbachia neither induces nor suppresses transcripts encoding antimicrobial peptides. Insect Mol Biol 9 635 639
24. DongY
AguilarR
XiZ
WarrE
MonginE
2006 Anopheles gambiae immune responses to human and rodent Plasmodium parasite species. PLoS Pathog 2 e52
25. Gonzalez-CeronL
SantillanF
RodriguezMH
MendezD
Hernandez-AvilaJE
2003 Bacteria in midguts of field-collected Anopheles albimanus block Plasmodium vivax sporogonic development. J Med Entomol 40 371 374
26. DongY
ManfrediniF
DimopoulosG
2009 Implication of the mosquito midgut microbiota in the defense against malaria parasites. PLoS Pathog 5 e1000423
27. PumpuniCB
BeierMS
NataroJP
GuersLD
DavisJR
1993 Plasmodium falciparum: inhibition of sporogonic development in Anopheles stephensi by Gram-negative bacteria. Exp Parasitol 77 195 199
28. PumpuniCB
DemaioJ
KentM
DavisJR
BeierJ
1996 Bacterial population dynamics in three anopheline species: the impact on Plasmodium sporogonic development. Am J Trop Med Hyg 54 214 218
29. NodaH
MiyoshiT
KoizumiY
2002 In vitro cultivation of Wolbachia in insect and mammalian cell lines. In Vitro Cell Dev Biol Anim 38 423 427
30. O'NeillSL
PettigrewMM
SinkinsSP
BraigHR
AndreadisTG
1997 In vitro cultivation of Wolbachia pipientis in an Aedes albopictus cell line. Insect Mol Biol 6 33 39
31. FurukawaS
TanakaK
FukatsuT
SasakiT
2008 In vitro infection of Wolbachia in insect cell lines. Appl Entomol and Zool 43 519 525
32. BarrettT
TroupDB
WilhiteSE
LedouxP
RudnevD
2009 NCBI GEO: archive for high-throughput functional genomic data. Nucl Acids Res 37 D885 890
33. XiZ
GavotteL
XieY
DobsonSL
2008 Genome-wide analysis of the interaction between the endosymbiotic bacterium Wolbachia and its Drosophila host. BMC Genomics 9 1
34. BrennanLJ
KeddieBA
BraigHR
HarrisHL
2008 The endosymbiont Wolbachia pipientis induces the expression of host antioxidant proteins in an Aedes albopictus cell line. PLoS ONE 3 e2083
35. FederME
KarrTL
YangFW
HoekstraJ
JamesDC
1999 Interaction of Drosophila and its endosymbiont Wolbachia: natural heat shock and the overcoming of sexual incompatibility. Amer Zool 39 363 373
36. LefevreT
ThomasF
SchwartzA
LevashinaE
BlandinS
2007 Malaria Plasmodium agent induces alteration in the head proteome of their Anopheles mosquito host. Proteomics 7 1908 1915
37. SimC
HongYS
TsetsarkinKA
VanlandinghamDL
HiggsS
2007 Anopheles gambiae heat shock protein cognate 70B impedes O'nyong-nyong virus replication. BMC Genomics 8 231
38. HaywardR
2000 Plasmodium falciparum phosphoenolpyruvate carboxykinase is developmentally regulated in gametocytes. Mol Biochem Parasitol 107 227 240
39. LasonderE
JanseCJ
van GemertG-J
MairGR
VermuntAMW
2008 Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity. PLoS Pathog 4 e1000195
40. del Pilar CorenaM
VanEkerisL
SalazarMI
BowersD
FiedlerMM
2005 Carbonic anhydrase in the adult mosquito midgut. J Exp Biol 208 3263 3273
41. ReungprapavutS
KrungkraiSR
KrungkraiJ
2004 Plasmodium falciparum carbonic anhydrase is a possible target for malaria chemotherapy. J Enzyme Inhib Med Chem 19 249 256
42. KrungkraiSR
SuraveratumN
RochanakijS
KrungkraiJ
2001 Characterisation of carbonic anhydrase in Plasmodium falciparum. Int J Parasitol 31 661 668
43. AdiniA
WarburgA
1999 Interaction of Plasmodium gallinaceum ookinetes and oocysts with extracellular matrix proteins. Parasitology 119 331 336
44. ArrighiRBG
LycettG
MahairakiV
Siden-KiamosI
LouisC
2005 Laminin and the malaria parasite's journey through the mosquito midgut. J Exp Biol 208 2497 2502
45. GareDC
PiertneySB
BillingsleyPF
2003 Anopheles gambiae collagen IV genes: cloning, phylogeny and midgut expression associated with blood feeding and Plasmodium infection. Int J Parasitol 33 681 690
46. WarburgA
ShternA
CohenN
DahanN
2007 Laminin and a Plasmodium ookinete surface protein inhibit melanotic encapsulation of Sephadex beads in the hemocoel of mosquitoes. Microb Infect 9 192 199
47. KumarS
ChristophidesGK
CanteraR
CharlesB
HanYS
2003 The role of reactive oxygen species on Plasmodium melanotic encapsulation in Anopheles gambiae. Proc Natl Acad Sci USA 100 14139 14144
48. BlandinSA
LevashinaEA
2004 Thioester-containing proteins and insect immunity. Mol Immunol 40 903 908
49. BlandinSA
ShiaoS-H
MoitaLF
JanseCJ
WatersAP
2004 Complement-like protein TEP1 is a determinant of vectorial capacity in the malaria vector Anopheles gambiae. Cell 116 661 670
50. LevashinaEA
MoitaLF
BlandinSA
VriendG
LagueuxM
2001 Conserved role of a complement-like protein in phagocytosis revealed by dsRNA knockout in cultured cells of the mosquito, Anopheles gambiae. Cell 104 709 718
51. VolzJ
MuellerH
ZdanowiczA
KafatosFC
OstaMA
2006 A genetic module regulates the melanization response of Anopheles to Plasmodium. Cell Microbiol 8 1392 1405
52. DimopoulosG
RichmanA
MüllerHM
KafatosFC
1997 Molecular immune responses of the mosquito Anopheles gambiae to bacteria and malaria parasites. Proc Natl Acad Sci USA 94 11508 11513
53. PaskewitzSM
AndreevO
ShiL
2006 Gene silencing of serine proteases affects melanization of Sephadex beads in Anopheles gambiae. Insect Biochem Mol Biol 36 701 711
54. AbrahamEG
PintoSB
GhoshA
VanlandinghamDL
BuddA
2005 An immune-responsive serpin, SRPN6, mediates mosquito defense against malaria parasites. Proc Natl Acad Sci USA 102 16327 16332
55. ZouZ
ShinSW
AlvarezKS
BianG
KokozaV
2008 Mosquito RUNX4 in the immune regulation of PPO gene expression and its effect on avian malaria parasite infection. Proc Natl Acad Sci USA 105 18454 18459
56. RichmanAM
DimopoulosG
SeeleyD
KafatosFC
1997 Plasmodium activates the innate immune response of Anopheles gambiae mosquitoes. EMBO J 16 6114 6119
57. WarrE
DasS
DongY
DimopoulosG
2008 The Gram-negative bacteria-binding protein gene family: its role in the innate immune system of Anopheles gambiae and in anti-Plasmodium defence. Insect Mol Biol 17 39 51
58. WernerT
LiuG
KangD
EkengrenS
SteinerH
2000 A family of peptidoglycan recognition proteins in the fruit fly Drosophila melanogaster. Proc Natl Acad Sci USA 97 13772 13777
59. PerssonC
OldenviS
SteinerH
2007 Peptidoglycan recognition protein LF: a negative regulator of Drosophila immunity. Insect Biochem Mol Biol 37 1309 1316
60. Zaidman-RémyA
HervéM
PoidevinM
Pili-FlouryS
KimM-S
2006 The Drosophila amidase PGRP-LB modulates the immune response to bacterial infection. Immunity 24 463 473
61. MeisterS
AgianianB
TurlureF
RelógioA
MorlaisI
2009 Anopheles gambiae PGRPLC-mediated defense against bacteria modulates infections with malaria parasites. PLoS Pathog 5 e1000542
62. ChoeK-M
WernerT
StövenS
HultmarkD
AndersonKV
2002 Requirement for a peptidoglycan recognition protein (PGRP) in Relish activation and antibacterial immune responses in Drosophila. Science 296 359 362
63. CarlssonA
EngströmP
PalvaET
BennichH
1991 Attacin, an antibacterial protein from Hyalophora cecropia, inhibits synthesis of outer membrane proteins in Escherichia coli by interfering with omp gene transcription. Infect Immun 59 3040 3045
64. SakamotoJM
AzadAF
2007 Propagation of arthropod-borne Rickettsia spp. in two mosquito cell lines. Appl Environ Microbiol 73 6637 6643
65. LiY
MaW-M
DaiJ-Q
FengC-Z
YangF
2008 Inhibition of a novel sperm gelatinase in prawn sperm by the male reproduction-related Kazal-type peptidase inhibitor. Mol Reprod Dev 75 1327 1337
66. LinM-H
LeeRK-K
HwuY-M
LuC-H
ChuS-L
2008 SPINKL, a Kazal-type serine protease inhibitor-like protein purified from mouse seminal vesicle fluid, is able to inhibit sperm capacitation. Reproduction 136 559 571
67. PerrimonN
EngstromL
MahowaldAP
1984 Developmental genetics of the 2E-F: a regions of the Drosophila X chromosome: a region rich in “developmentally important” genes. Genetics 108 559 572
68. BurnetteJM
HattonAR
LopezAJ
1999 Trans-acting factors required for inclusion of regulated exons in the Ultrabithorax mRNAs of Drosophila melanogaster. Genetics 151 1517 1529
69. ChungS
ZhouZ
HuddlestonKA
HarrisonDA
ReedR
2002 Crooked neck is a component of the human spliceosome and implicated in the splicing process. Biochim Biophys Acta 1576 287 297
70. JiangX
XiaL
ChenD
YangY
HuangH
2008 Otefin, a nuclear membrane protein, determines the fate of germline stem cells in Drosophila via interaction with Smad complexes. Dev Cell 14 494 506
71. VardanyanA
AtanesyanL
EgliD
RajaSJ
Steinmann-ZwickyM
2008 Dumpy-30 family members as determinants of male fertility and interaction partners of metal-responsive transcription factor 1 (MTF-1) in Drosophila. BMC Dev Biol 8 68
72. DottoriniT
NicolaidesL
RansonH
RogersDW
CrisantiA
2007 A genome-wide analysis in Anopheles gambiae mosquitoes reveals 46 male accessory gland genes, possible modulators of female behavior. Proc Natl Acad Sci USA 104 16215 16220
73. SnookRR
ClelandSY
WolfnerMF
KarrTL
2000 Offsetting effects of Wolbachia infection and heat shock on sperm production in Drosophila simulans: analyses of fecundity, fertility and accessory gland proteins. Genetics 155 167 178
74. TurleyAP
MoreiraLA
O'NeillSL
McGrawEA
2009 Wolbachia Infection Reduces Blood-Feeding Success in the Dengue Fever Mosquito, Aedes aegypti. PloS Negl Trop Dis 3 e516
75. MoreiraLA
SaigE
TurleyAP
RibeiroJMC
O'NeillSL
2009 Human probing behavior of Aedes aegypti when infected with a life-shortening strain of Wolbachia. PloS Negl Trop Dis 3 e568
76. GoldmanTD
ArbeitmanMN
2007 Genomic and functional studies of Drosophila sex hierarchy regulated gene expression in adult head and nervous system tissues. PLoS Genet 3 e216
77. BudanovAV
SablinaAA
FeinsteinE
KooninEV
ChumakovPM
2004 Regeneration of peroxiredoxins by p53-regulated sestrins, homologs of bacterial AhpD. Science 304 596 600
78. LeeJH
BudanovAV
ParkEJ
BirseR
KimTE
2010 Sestrin as a feedback inhibitor of TOR that prevents age-related pathologies. Science 327 1223 1228
79. MinKT
BenzerS
1997 Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death. Proc Natl Acad Sci USA 94 10792 10796
80. PfafflMW
2001 A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29 e45
81. WaterhouseRM
KriventsevaEV
MeisterS
XiZ
AlvarezKS
2007 Evolutionary dynamics of immune-related genes and pathways in disease-vector mosquitoes. Science 316 1738 1743
82. DanaAN
HongYS
KernMK
HillenmeyerME
HarkerBW
2005 Gene expression patterns associated with blood-feeding in the malaria mosquito Anopheles gambiae. BMC Genomics 6 5
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