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Immune response and
disease tolerance in shellfish

              Steven Roberts
             University of Washington
    ...
flickr | Basenisa
Stress
•   Harmful anthropogenic compounds
•   Temperature

•   Hypoxia

•   Harmful organisms
      Bacteria,Viruses, Par...
Sam White
Innate Immune System

• Present from birth
• “Non-specific” - acts on many pathogens
  and does not show specificity within ...
Laura Canesi, Gabriella Gallo, Miriam Gavioli, and Carla Pruzzo
Bacteria–hemocyte Interactions and Phagocytosis in Marine
...
flickr | cpurrin1
Outline

• Genes expressed in oyster hemocytes
• Dermo disease and mechanical stress
• Disease resistance in oysters (and ...
cDNA Library




               Rick Goetz
Interleukin-17
gene expression level
(fold increase over minimum)
Dermo disease and mechanical stress




Maps: Google - Oyster photos: G. Burreson
Dermo disease and mechanical stress




                          Gary Wikfors and Inke Sunila
Stress and Disease
Resistance ?
MVineyard                   flickr | cpurrin1




               Roxannna Smolowitz
                  Rick Karney
Another example




          Dale Leavitt, Marta Gomez-Chiarri
Markers ?
Glenn VanBlaricom, Carolyn Friedman
Abalone
Glenn VanBlaricom, Carolyn Friedman
Carolyn Friedman
Mining ESTs
                         from similar species
Gene                    Molecular Function         Biological Pr...
TOLLIP   TOLLIP
Clams




        Roxanna Smolowitz
Differential Display
Serine Protease Inhibitor

• Critical factor in disease pathogenesis

• Serine proteases produced by pathogen
  •   Facili...
Expression
0.009
0.008
0.007
0.006
0.005
0.004
0.003
0.002
0.001
   0
        MA +   MA -    FL +            FL -   NJ +  ...
photo: Maille Lyons
Field Trials
                   100
                                     June
                                     August
...
QPX gene expression
QPX Serine Protease
Top Blastx Hit                                    Species associated with Blast Hit
                  ...
Relative Gene Expression
Relative Gene Expression




                                                                    ...
thanks




         Smolowitz
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
Shellfish Disease
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Shellfish Disease

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Shellfish Disease

  1. 1. Immune response and disease tolerance in shellfish Steven Roberts University of Washington School of Aquatic and Fishery Sciences
  2. 2. flickr | Basenisa
  3. 3. Stress • Harmful anthropogenic compounds • Temperature • Hypoxia • Harmful organisms Bacteria,Viruses, Parasites
  4. 4. Sam White
  5. 5. Innate Immune System • Present from birth • “Non-specific” - acts on many pathogens and does not show specificity within a pathogen type • Does not become more efficient on subsequent exposure to same pathogen
  6. 6. Laura Canesi, Gabriella Gallo, Miriam Gavioli, and Carla Pruzzo Bacteria–hemocyte Interactions and Phagocytosis in Marine Bivalves. 2002. Microscopy Research And Technique 57:469–476
  7. 7. flickr | cpurrin1
  8. 8. Outline • Genes expressed in oyster hemocytes • Dermo disease and mechanical stress • Disease resistance in oysters (and abalone) • Clams and QPX
  9. 9. cDNA Library Rick Goetz
  10. 10. Interleukin-17
  11. 11. gene expression level (fold increase over minimum)
  12. 12. Dermo disease and mechanical stress Maps: Google - Oyster photos: G. Burreson
  13. 13. Dermo disease and mechanical stress Gary Wikfors and Inke Sunila
  14. 14. Stress and Disease
  15. 15. Resistance ? MVineyard flickr | cpurrin1 Roxannna Smolowitz Rick Karney
  16. 16. Another example Dale Leavitt, Marta Gomez-Chiarri
  17. 17. Markers ?
  18. 18. Glenn VanBlaricom, Carolyn Friedman
  19. 19. Abalone
  20. 20. Glenn VanBlaricom, Carolyn Friedman
  21. 21. Carolyn Friedman
  22. 22. Mining ESTs from similar species Gene Molecular Function Biological Process Cellular Component rab1 GTPase activity vesicle-mediated golgi apparatus transport rab7 (WSSV receptor) GTPase activity, protein intracellular protein golgi apparatus, binding transport lysosome catalase catalase activity, response to oxidative peroxisome, plasma antioxidant activity stress membrane toll-interacting protein protein binding, signal inflammatory response cytoplasm transducer activity beta-glucan bacterial binding regulation of innate extracellular region recognition protein immune response manganese superoxide MnSD activity, response to oxidative mitochondrial matrix dismutase antioxidant activity stress plancitoxin deoxyribonuclease II DNA catabolic process, lysosome activity DNA binding Lindsay Braun and Cullen Taplin
  23. 23. TOLLIP TOLLIP
  24. 24. Clams Roxanna Smolowitz
  25. 25. Differential Display
  26. 26. Serine Protease Inhibitor • Critical factor in disease pathogenesis • Serine proteases produced by pathogen • Facilitate penetration • Stimulate degradation of host proteins involved in immune defense
  27. 27. Expression 0.009 0.008 0.007 0.006 0.005 0.004 0.003 0.002 0.001 0 MA + MA - FL + FL - NJ + NJ - Strain Clams from different location of origin (MA, FL, NJ) all grown in common field sites in MA. Plus (+) and minus (-) symbols indicate two locations within a harbor.
  28. 28. photo: Maille Lyons
  29. 29. Field Trials 100 June August Percent Infected 75 50 25 0 MA FL NJ
  30. 30. QPX gene expression
  31. 31. QPX Serine Protease Top Blastx Hit Species associated with Blast Hit e-value % similar Glutathione-dependent formaldehyde-activating, GFARalstonia metallidurans 1.00E-04 45 Subtilisin-like serine protease pepC precursor Aspergillus niger 2.00E-16 63 Permeases of the major facilitator superfamily Yersinia frederiksenii 0.66 47 40s ribosomal protein s23 Theileria annulata 9.00E-38 96 ATP-binding cassette / transporter, sub-family G, Toxoplasma gondii member 4 4.00E-25 60 mitogen-activated protein kinase Pan troglodytes 3.30E+00 46 Multidrug Resistance Protein family member (mrp-1)Caenorhabditis elegans 3.00E-14 66 mitogen-activated protein kinase 2 Toxoplasma gondii 1.00E-12 84 haemagluttinin repeat protein Pseudomonas fluoresens 2.70E+00 52 polysaccharide biosythesis protein Methylococcus capsulatus 4.30E+00 47 Potassium inwardly-rectifying channel Homo sapiens 1.00E-07 55
  32. 32. Relative Gene Expression Relative Gene Expression Optimal Clam Tissue Clam Tissue Media and Water and Media Culture Conditions (ATTC strain) ATTC ATTC Env_66 Env_66 21! 10! 21! 10! Strain / Temperature
  33. 33. thanks Smolowitz

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