1. Immune response and
disease tolerance in shellfish
Steven Roberts
University of Washington
School of Aquatic and Fishery Sciences

2. flickr | Basenisa

3. Stress
• Harmful anthropogenic compounds
• Temperature
• Hypoxia
• Harmful organisms
Bacteria,Viruses, Parasites

4. Sam White

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. 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. flickr | cpurrin1

8. Outline
• Genes expressed in oyster hemocytes
• Dermo disease and mechanical stress
• Disease resistance in oysters (and abalone)
• Clams and QPX

9. cDNA Library
Rick Goetz

11. Interleukin-17

15. gene expression level
(fold increase over minimum)

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

17. Dermo disease and mechanical stress
Gary Wikfors and Inke Sunila

19. Stress and Disease

20. Resistance ?
MVineyard flickr | cpurrin1
Roxannna Smolowitz
Rick Karney

26. Another example
Dale Leavitt, Marta Gomez-Chiarri

27. Markers ?

28. Glenn VanBlaricom, Carolyn Friedman

29. Abalone

30. Glenn VanBlaricom, Carolyn Friedman

31. Carolyn Friedman

32. 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

33. TOLLIP TOLLIP

34. Clams
Roxanna Smolowitz

37. Differential Display

38. Serine Protease Inhibitor
• Critical factor in disease pathogenesis
• Serine proteases produced by pathogen
• Facilitate penetration
• Stimulate degradation of host proteins involved
in immune defense

39. 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.

41. photo: Maille Lyons

42. Field Trials
100
June
August
Percent Infected
75
50
25
0
MA FL NJ

44. QPX gene expression

45. 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

46. 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

48. thanks
Smolowitz