1 / 29

Finalising the cytokine repertoire of salmonid fish Chris Secombes

Finalising the cytokine repertoire of salmonid fish Chris Secombes Scottish Fish Immunology Research Centre University of Aberdeen. Cytokine Network. Interleukins. Interferons. Tumor necrosis factors. Chemokines ( chemo attractive cyto kines ).

meryle
Download Presentation

Finalising the cytokine repertoire of salmonid fish Chris Secombes

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Finalising the cytokine repertoire of salmonid fish Chris Secombes Scottish Fish Immunology Research Centre University of Aberdeen

  2. Cytokine Network Interleukins Interferons Tumor necrosis factors Chemokines (chemoattractive cytokines)

  3. Attempts to discover cytokine genes in • fish have used: • Homology cloning • EST analysis • Genome analysis • All have had some degree of success

  4. Sequenced Fish Genomes Medaka Fugu Stickleback Tetraodon Zebrafish Cod

  5. MDM1 MDM1 MDM1 MDM1 DYRK2 DYRK2 DYRK2 DYRK2 IL-26rel IL-26rel IL-26rel RAP1B RAP1B RAP1B RAP1B IFN- IFN- IFN- IFN- IL-22 IL-22 IL-22 IL-22 Human IL-26 Fugu Stickleback Zebrafish IFN- locus IFN-: ~35-40% aa homology to mammals ~50% aa homology between fish

  6. Teleost fish IL-6 IL-11a IL-34 IL-2 IL-11b MCSF1 IL-2-rel G-CSF MCSF2 IL-4/13A CNTF-like SCF IL-4/13B M17 IL-7 IL-12 IL-15 IL-23 IL-15L IL-35 IL-21 IL-17A/F1 IL-17A/F2 GDF15 IL-17A/F3 TGF-1 IL-17C/E TGF-2 IL-17D TGF-3 IL-17N TGF-6 IL-10 IFNa IL-19/20/24 IFNb IL-22 IFNc IL-26? IFNd IFN- IFNe IFN--relIFNf IL-1I IL-1II IL-18 nIL-1F CXCL1-25 CCL1-81 XCL1 CXL1-4 TNF-I TNF-II LT-? Type II Type I IL-10 IFN IL-19 IFN IL-20 IFN IL-22 IFN IL-24 IFN IL-26 IFN IFN- IFNL1 IFNL2 IFNL3 CXCL1-16 CCL1-28 CX3CL1 XCL1-2 IL-6 IL-12 IL-2 IL-11 IL-23 IL-3 OSM IL-27 IL-4 LIF IL-35 IL-5 CNTF IL-7 CLC IL-34 IL-9 CT1 MCSF IL-13 CT2 GMCSF IL-15 GCSF SCF IL-21 IL-1F1-13 IL-17A TGF-1 IL-17B TGF-2 IL-17C TGF-3 IL-17D GDF15 IL-17E IL-17F TNF- LT- LT- Mammals

  7. MDM1 MDM1 MDM1 MDM1 DYRK2 DYRK2 DYRK2 DYRK2 IL-26rel IL-26rel IL-26rel RAP1B RAP1B RAP1B RAP1B IFN- IFN- IFN- IFN- IL-22 IL-22 IL-22 IL-22 IFN-rel IFN-rel Human IL-26 Fugu Zebrafish IFN- locus Stickleback IFN-rel

  8. Signature Motif IFN-rel: A second type II IFN in fish

  9. Signature Motif Nuclear localisation signal IFN-rel: A second type II IFN in fish

  10. Trout IL-15 BCL10 INPP4B Bei et al. MolecImmunol 43, 2006, 860-869.

  11. Innate immune cells Adaptive immune cells / IEL NK Cell NK T Cell CD8+ CD4+ Dependence on IL-15 signals IL-15 signals differentially support immune cells

  12. Human Chromosome 5 143 Mb 130 Mb 135 Mb YIPF5 SPRY4 RNF14 ARHGAP26 NR3C1 FGF-1 UQCRQ SHROOM1 FBXL21 NDFIP1 IL-9 SLC22A5 LECT2 KIF3A ANKRD43 SEPT8 CNNI PDLIM4 SLC22A4 IL-4 IL-5 AFF4 GDF9 IL-13 IRF-1 HSPA4 P4HA2 RAD50 LEAP-2 Stickleback Tetraodon Fugu Genome UQCRQ SEPT8 GDF9 NUDCD2 CCNG1 NDFIP1 FGF1 NR3C1 SH3RF2 FGF16 ZDHHC15 UPRT KCTD16 HSPA4 GNPDA1 ANKRD43 KIF3A IL-4/13B ATRX ABCB7 GABRB2 GABRA6 Stickleback Tetraodon Fugu Genome GPR112 FHL1 SLC9A6 DRD1 RNF14 NR3C1 YIPF5 GRXCR2 POU4F3 MSX2 HMP19 NDFIP1 SPRY4 FGF1 ARHGAP26 KCTD16 RBM27 RAD50 CPEB4 LARS IL-4/13A PLAC8L1 Th2 cytokines Li et al, Molec Immunol 44, 2007, 2078-2086.

  13. Family No. of species Gar Lepisosteida Outgroup Sardine Clupeomorpha 357 Zebrafish Carp Ostariophysi 6375 Catfish Salmon Protacanthopterygii 312 Trout Cod Paracanthopterygii 1212 MedakaAtherinomorpha 1283 Halibut Pleuronectiformes 570 Stickleback Gasterosteiformes 257 Tilapia Perciformes 9293 Fugu Tetraodontiformes 339 Tetraodon Davidson et al., Genome Biology 11, 2010, 403.  

  14. TNF, IL-1, Chemokines (eg IL-8)

  15. 89 138 397 intron (bp) TNF1 154 192 49 60 440 503 exon (bp) 191 219 49 60 440 590 exon (bp) TNF2 89 138 395 intron (bp) AT rich repeats Time (hrs) after LPS 140 0 2 4 8 24 36 120 100 TNF1 80 No. of Leucocytes per Field of View (x400) 60 40 TNF2 20 0 PBS 1ng 2ng 10ng 20ng 100ng 200ng Treatment/Dose b-actin TNF1 TNF2 Two TNF genes in trout 88% amino acid identity

  16. Teleost TNFSF Human TNFSF LIGHT-v1 4-1BBL CD27-L LIGHT 4-1BBL Chr 19 Tn Chr 18 (Dr Chr 3) ? Notch-3 1.5kb 49kb 73kb FasL FAS-L Ox40L GITRL ? ? Dr Chr 20 Chr 1 Notch-2 132kb 372kb HumanMHC-Paralogousregions TL1A-like CD30L TL1A ? Dr Chr 8 Chr 9 Notch-1 TNF-a v2 96kb TNF-N LT-b TNFa LT-a ? Dr Chr 15 19kb Chr 6 TNF-a v1 Notch-4 2kb 3.2kb Dr Chr 19 EDA CD40L CD40L BALM EDA Chr X Tn Chr 1 (Dr Chr 14) ? 6.8Mb 1kb 55Mb TRAIL-like-v4 APRIL APRIL TWEAK Dr Chr 5 Chr 17 ? 39Mb 707bp RANKL BAFF BAFF RANKL Tn Chr 2 (Dr Chr 9) 3.0Mb ? Chr 13 TRAIL-like-v3 65Mb Dr Chr 24 TRAIL TRAIL-like-v2 TWEAK Chr 3 Dr Chr 7 19Mb TRAIL-like-v1 Dr Chr 21 ? Glenney & Wiens, J Immunol 178, 2007, 7955-73.

  17. HorseIL18 0.1 PigIL18 CowIL18 DogIL18 IL-18 HumanIL18 RabbitIL18 MouseIL18 DuckIL18 ChickenIL18 TroutIL18 FuguIL18 IL-33 HumanIL33 DogIL33 MouseIL33 CowIL1a PigIL1a HorseIL1a IL-1 DogIL1a DolphinIL1a HumanIL1a RabbitIL1a MouseIL1a nIL-1F Trout nIL1F Tetraodon nIL1F CowIL1ra DolphinIL1ra PigIL1ra IL-1ra DogIL1ra HumanIL1ra HorseIL1ra RabbitIL1ra MouseIL1ra HumanIL1F5 MouseIL1F5 HumanIL1F10 MouseIL1F10 IL-1F6-10 HumanIL1F6 MouseIL1F6 HumanIL1F9 MouseIL1F9 HumanIL1F8 MouseIL1F8 HumanIL1F7 HalibutIL1b TurbotIL1b CobiaIL1b IcefishIL1b ChineseperchIL1b SeabreamIL1b RedbreamIL1b SeabassIL1b TetraodonIL1b SalmonIL1b TroutIL1b1 TroutIL1b2 CodIL1b HaddockIL1b CarpIL1b21 CarpIL1b22 IL-1 GoldfishIL1b1 GoldfishIL1b2 CarpIL1b1 ZebrafishIL1b MouseIL1b RabbitIL1b HumanIL1b HorseIL1b DogIL1b CowIL1b DolphinIL1b PigIL1b PossumIL1b PlatypusIL1b XlaevisIL1b ChickenIL1b CatsharkIL1b LeopardsharkIL1b The IL-1 cytokine family

  18. Cytokines produced/ protective activity Th1-cells IL-2, IFN-, LT- Intracellular bacteria and some viruses Th2-cells IL-4, IL-5, IL-9, IL-13, IL-20, IL-31 Helminths IL-17A, IL-17F, IL-22 Th17-cells Extracellular bacteria and some fungi Wilson et al, Nature Reviews Immunology 9, 2009, 91-105.

  19. Human IL-17A 57 House IL-17A 73 Cow IL-17A 99 Pig IL-17A 84 Mouse IL-17A 57 99 Rat IL-17A Human IL-17F IL-17A/F 98 Rat IL-17F 99 Mouse IL-17F 99 Anolis IL-17A Duck IL-17A 54 97 Chicken IL-17A 100 Chicken IL-17F 99 96 Zebrafish IL-17A/F-1 Zebrafish IL-17A/F-3 Zebrafish IL-17A/F-2 40 99 Trout IL-17A/F 99 Mouse IL-17B IL-17B 71 Human IL-17B Guttata IL-17B 99 75 Chicken IL-17B Frog IL-17B 73 Frog IL-17D2 73 Lamprey IL-17 99 Salmon IL-17D 99 Trout IL-17D 93 IL-17D Zebrafish IL-17D Frog IL-17D1 99 Guttata IL-17D 80 Mouse IL-17D 76 Human IL-17D 99 Fish IL-17C 98 Zebrafish IL-17C 76 Promelas IL-17C 97 Catfish IL-17C Trout IL17C1 Trout IL17C2 99 IL-17C Guttata IL-17C 98 91 99 Mouse IL-17C Rat IL-17C Dog IL-17C 99 Human IL-17C 79 82 IL-17E Mouse IL-17E Human IL-17E 99 Horse IL-17E 87 0.2 Cow IL-17E 80 IL-17 cytokine family in fish Considered a Th2 cytokine

  20. IL-17 loci in Zebrafish Gunimaladevi et al, Fish & Shellfish Immunol 21, 2006, 393-403.

  21. IL-17 loci in Fugu Korenaga et al, Fish & Shellfish Immunol 28, 2010, 809-818.

  22. Chemokines

  23. 111 chemokine genes now identified in zebrafish (v6 assembly). 25 CXC, 81 CC, 4 CX and 1 XC. But - only 18 genes found in pufferfish Nomiyama et al, BMC Genomics 9, 2008, 222.

  24. CXCL1-8 CXCL13 Nomiyama et al, BMC Genomics 9, 2008, 222.

  25. Interferons

  26. zIFN1 zIFN2 Chromosome 3 18,200,000 START START STOP STOP STOP START zIFN3 Chromosome 12 5,221,000 START STOP zIFN4 Salmon IFN genes In zebrafish, four IFN genesare located in two clusters on Chr 3 and 12 1 kb Group II IFNs (two C-C pairs) Group I IFNs (one C-C pair) Aggad et al, J. Immunol 183, 2009, 3924-31. Sun et al, Dev Comp Immunol 33, 2009, 547-58.

  27. Mammals/ Birds Fugu Zebrafish Salmon Trout IFN-1 and 2 Interferon gene organisation Presence of introns in fish IFN genes allows alternative splicing.

  28. Conclusions We will soon know the major players within the salmonid cytokine network. In some cytokine families there will be a large number of genes to be analysed functionally. Expression analysis will help identify the most important paralogues to study functionally. However, it is still going to be many years of further study before we really have a good understanding of their function within the salmonid immune system.

  29. Imaquanim programme Thanks to: Dr Steve Bird* Dr Suhee Hong Mansourah Husain Dr Sam Martin Dr Tiehui Wang Dr Jun Zou Bertrand Collet, MSS, Aberdeen, UK Pierre Boudinot & Carine Genet INRA, France MingXian Chang CAS, China *Now at: Waikato University, NZ

More Related