C. elegans as an. Centre d Immunologie de Marseille-Luminy

Transcription

1 C. elegans as an alternative ti model host Centre d Immunologie de Marseille-Luminy

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4 Why C. elegans? Small size Transparent Rapid generation time evgen nl De e taken from h image 0.1 mm

5 46 hr at 20 C

6 Virulence factors

7 Identity of P. aeruginosa mutants Gene Gene product gaca Two-component regulator gacs Two-component regulator lasr Quorum-sensing regulator ptsp mtrr aefa X X Transcriptional regulator Transcriptional regulator of multi-drug transporter Integral membrane protein Unknown Unknown Tan and Ausubel (2000) Curr. Op. Microbio.

8 Universal virulence factors Screen in C. elegans 8 mutants Decreased virulence in: Arabidopsis 6 Wax moth 6 7 Mahajan-Miklos et al. (2000) Mol. Microbiol.

9 E. coli is broken down in the pharynx Kurz et al. (2003) Embo J Bacteria Autofluorescent gut granules Short contact with fluorescent E. coli

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11 S. marcescens destroys the grinder Short contact with fluorescent S. marcescens Kurz et al. (2003) Embo J

12 Screening Bank of S. marcescens Tn5 mutants Worms Individual clones 96 well plates Infection Less virulent clones

13 Attenuated mutants SM202 20C2 SM204 SM237 wbdc wbdb wbda wbbd wzt wzm manb1manc1 gale manb2manc2 O-antigen biosynthesis

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16 Host response

17 5 Transcriptional response to Photorhabdus luminescens Genes

18 5 Transcriptional response to P. luminescens compared to Db Genes

19 Host response to infection is specific Wong et al. (2007) Genome Biol.

20 Host response to infection includes a common signature Wong et al. (2007) Genome Biol.

21 Infection by P. luminescens induces asp-4::gfp in the intestine ti DIC GFP fluorescence necrotic cells necrotic GFP expression cells Wong et al. (2007) Genome Biol.

22 Suppression of necrosis increases resistance to infection E. carotovora Wong et al. (2007) Genome Biol.

23 Impossible d'afficher l'image. Votre ordinateur manque peut-être de mémoire pour ouvrir l'image ou l'image est endommagée. Redémarrez l'ordinateur, puis ouvrez à nouveau le fichier. Si le x rouge est toujours affiché, vous devrez peut-être supprimer l'image avant de la réinsérer. Drechmeria coniospora spores adhere to the nematode cuticle Jürgen Berger, MPI, Tübingen.

24 Impossible d'afficher l'image. Votre ordinateur manque peut-être de mémoire pour ouvrir l'image ou l'image est endommagée. Redémarrez l'ordinateur, puis ouvrez à nouveau le fichier. Si le x rouge est toujours affiché, vous devrez peut-être supprimer l'image avant de la réinsérer. Drechmeria coniospora spores adhere to the nematode cuticle Jürgen Berger, MPI, Tübingen.

25 nlp peptide genes are induced rms alive % wo50 Microarrays Antimicrobial peptide p genes Time (Hours) Couillault et al. (2004) Nat. Immun.

26 Induction of nlp-29 after Drechmeria infection Pathogen Is7 transgenic strain pcol-12 RFP constitutive pnlp-29 GFP inducible Non-infected Infected

27 Induction of pnlp-29::gfp Gree en fluo orescen nce Infected Non-infected n > 1,000 Length

28 In vivo identification of signaling pathways No infection Infection Infection A B C A B C A B C pcol-12l dsred pcol-12l dsred pcol-12l constitutive dsred pnlp-29 GFP inducible pnlp-29 GFP pnlp-29 GFP RED GREEN RED

29 TIR/MAPK pathway involved in defence FUNGAL INFECTION TIR-1 MAPKKK NSY-1 MAPKK SEK-1 MAPK PMK-1 ANTIMICROBIAL PEPTIDES

30 Blocking induction of pnlp-29::gfp Control tir-1 (RNAi) Couillault et al. (2004) Nat. Immun.

31 C. elegans is attracted by bacteria E. coli

32 C. elegans is attracted by bacteria E. coli

33 C. elegans prefers S. marcescens E. coli S. mar. PATHOGEN

34 C. elegans prefers S. marcescens E. coli S. mar. PATHOGEN

35 Worms are repelled by S. marcescens S. marcescens Db11

36 Worms are repelled by S. marcescens S. marcescens Db11

37 Worms are not repelled by SM267 S. marcescens SM267 Pradel et al. (2007) PNAS

38 SM267 contains an insertion in the swra gene that t encodes a NRPS Condensation domain AMP-binding domain Phosphopantetheine attachment Thioesterase domain Pradel et al. (2007) PNAS

39 swra is necessary for the synthesis of serrawettin W2 L-Ser D-Leu 3-hydroxydecanoic acid H HO H L-Thr O H IN N I H O OH H N H H O O N I H H H H I N H O O H O L-Ile D-Phe Pradel et al. (2007) PNAS

40 Conclusions C. elegans can be used to identify virulence factors can be used to identify antimicrobials responds specifically to infection complex control at the genetic and cellular level detects pathogens

41 Equipe FRM C. Couillault S. Cypowyj Y. Duverger L. Kurz K.Z. Lee L. Matthews N. Pujol D. Wong K. Ziegler O. Zugasti Tohey Matsuyama Niigata Cori Bargmann New York Elizabeth Pradel Centre d Immunologie de Marseille-Luminy

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43 Known pathogens of C. elegans Gram-positive bacteria: Bacillus thuringiensis Enterococcus faecalis Listeria monocytogenes Microbacterium nematophilum Staphylococcus aureus Streptococcus t pneumoniae Fungi: Gram-negative bacteria: Acinetobacter baumannii Aeromonas hydrophila Agrobacterium tumefaciens Burkholderia cepacia B. thailandensis B. pseudomallei Erwinia christamthemi E. carotovora carotovora Escherichia coli (EPEPC & EHEC) Arthrobotrys sp. Candida albicans Cryptococcus neoformans Drechmeria coniospora Duddingtonia flagrans Paecilomyces lilacinus i Pseudomonas aeruginosa P. fluorescens Salmonella typhimurium Shewanella frigidimarina S. massalia Serratia marcescens Vibrio cholerae Yersinia spp. VIRUSES