™Fish are the most primitive vertebrates to

posses adaptive immune system (lymphocytes,

Ig, MHC and TCR)

™Fish immune system is simple and

undifferentiated.
Comparison of vertebrate immune system
 Immune Tissues and Organs

I
Immunocompetent
t t organs:

¾Thymus: Th, Tc-cells

¾Kidney: undifferentiated blood cell

¾Spleen: lymphocytes and macrophages

¾Liver: complement cascade

 Immune system classification:

‰Innate VS Adaptive immune

response

‰Non-specific VS specific immune

response
Hematopoietic
p stem cells produce
p cells
in blood and lymph

Adapted from Biology of

the Immune System,
JAMA 278
 Fish Immune System
y
Innate/Natural Immunity
mm y
an inherited, non-specific immunity
Resistance
mucus layer: physical and chemical
barrier (bactericidal substance)
Acquired
q Immunity
y
It was stimulated by contact with the
antigens.
Adaptive immunity
‰T cells (T cell receptors and MHC)
‰B cells
ll (I
(Immunoglobulin)
l b li )
Innate immunity
‰body surface (scale, mucus, etc)
‰macrophage
‰interferon
‰inflammation
‰serum proteins (lection, transferrin, CRP, etc)
‰complement component
 Fish Immune System

Nonspecific defense mechanisms

¾ Behavior aspects
¾ Physical barrier: Skin, scales, and mucus
¾ Chemical defenses: mucus, Lysozyme, Ig,
Fr fatty
Free f tt acids,
cids C’
C.
¾ Humoral factors
¾ Cellular factors
 Fish Immune System

Specific defense mechanisms

¾ H
Humoral
l iimmune system:
t I
Ig

¾ C ll
Cell-mediated
di d iimmune system: L
Leukocytes
k
 Nonspecific:
p Behavioral aspects
p

• Swimming behavior
– Flashing
g
– Out of schooling
– Settle down on the bottom
 Nonspecific : physical barrier

• Epithelium barrier
– Non-keratinized
– Wound
W dhhealing
li (outsideÆ
( t id Æ inside)
i id )
(mammals: inside Æ outside of the wound)
(mammals
• Mucus
– Goblet cells: response to infection, physical or
chemical irritant or pollutants
• Lysozyme, C-reactive proteins, Complement,
Protease, hemagglutinins and lipids
 Nonspecific : Humoral factors

• Acute phase reactants

–C
C-reactive
reactive protein

– Ceruloplasmin

– Transferrin
• Lectins and lysins

• Complement (C)
 Non-specific Humoral Molecules:
Molecule Composition Mode of Action
Lectins Specific sugar-binding Recognition,
proteins
p p
precipitation,
p
agglutination
Lytic Catalytic proteins Hemolytic and
enzymes lysozyme, etc. antibacterial activity
Transferrin/ Glycoprotein Iron binding
l
lactoferrin
f
Ceruloplasmin Acute-phase protein Copper binding
C-reactive Acute-phase protein Activation of
protein complement
Interferon protein Resistance to viral
infection
 Overview of the complement cascade
Specific
f antibody
b d

P th
Pathogen

Complement activation

Classical p
pathway
y Lectin p
pathway
y Alternative p
pathway
y
Antigen:antibody Lectin binding to
Pathogen surfaces
complex pathogen surfaces

Complement activation

Recruitment
R it t of
f Opsonization
O i ti of f Killing of
Killi f pathogens
th
inflammatory cells pathogens (Membrane attack complex, lysis of
cells)
 Nonspecific: Cellular defenses

• Macrophage and mononuclear phagocytes

• Melanin-containing cells

– absorb free radicals and reduce tissue damage

• Granulocytes

– Neutrophil: Phagocytosis!

– Eosinophilic granular cells (EGCs)

• Nonspecific Cytotoxic Cells (NCC)

 Leukocyte responses of fish to
noninfectious and infectious diseases
Species Stressor Total leukocytes Lymphocytes

Channel catfish Transport Ð Ð

Rainbow trout Surgery

g y Ð

Brown trout Handling Ð

Rainbow trout Pulp mill effluent Ð

American eel Cadmium Ï

Goldfish Aeromonas Ð
h d hil
hydrophila
Salmonid Infectious Ð
h
hematopoetic
t ti
necrosis (IHN)
Nonspecific immunity: Cellular defenses

• Inflammation
– No pus formation, liquefaction

• Granulomatous inflammation

– Epitheloid cells, high metabolic

activity of phagolysosomes

– Cytoplasmic interdigitation between

cells
 Specific Immunity

• Specific
S ifi response tto antigenic
ti i stimulation
ti l ti

• Lymphoid organs:

– Primary : thymus, kidney

– Secondary : spleen, gut-associated and gonadal

lymphoid tissues
 Specific: CMI

‰ Relies
R li on contact
t t of f th
the f
foreign
i iinvader
d with
ith
the subsequent presentation of an antigen having
the same major histocompatability complex (MHC
I or II) to T
T-helper
helper cells
‰ Once T-helper cells are stimulated, the produce
cytokines
t ki th
thatt result
lt iin stimulation
ti l ti of f effecter
ff t
cells (cytotoxic lymphocytes) or macrophages
‰ Cytokines stimulate of ore mentioned cells and
also recruit new cells to the area,
area activate them
 Specific:
p f HMI
¾Leukocytes: B-cell,
B-cell T-cell

¾Thrombocyte

¾M t
¾Monocyte

G l
¾Granulocytes: neutrophil,
h l eosinophil,
h l basophil
b hl

¾Nonspecific cytotoxic cells

 Specific humoral immunity
Light
chain

• Antibody: Ig M, Ig D
– Structure: Disulfid
e bond

• Human Ig M = pentamer Heavy

H
chain
• Teleost Ig M = tetramer
– Functions:
• Neutralization
• Precipitation
• Opsonization
• Complement mediated reactions
Ig evolution
 Fish antibody
• Affinity variation of Ab
– Ig MÆ not increase affinity after booster
• Kinetics of Ab formation
– Long inductive phase
– Temperature
T t effects
ff t
• Decreased Ab diversity
y
• Distribution
– Serum Ab: found after infection,
infection prophylaxis
– Mucosal Ab: GI, skin
– Maternal transfer of Ab: in some spp.
spp
 Specific cellular immunity

• Delayed hypersensitivity
– Localized accumulation of lymphoid
cells
– Later stage of chronic inflam.
inflam
(mycobacteriosis)
• Allograft
All f immunity
i i
– Graft rejection
– Infiltrate lymphoid cells to the graft
 Specific immune response induced
by
b an antigen
ti bath
b th

Antigen (bacterin,
(bacterin vaccine)
Spleen,
Gills Circulation
kidney
Antibody
Survillance Phagocytic Lymphocytes
in serum
cells Transporting cells
monocytes
Activated T cells
B cells
Macrophages
Cytotoxic cells
Monocytes
Neutrophils
Conclusion of Fish Defenses
 Immune Response System

1) cell-mediated immunity (T cells) and

2) humoral antibody system (B cells)
Response of Fish Following an Encounter
with
ith a Pathogen
P th
Fish Contacts Pathogen

I
Innate
t IImmunity
it
Failure (Disease Success (No
and Death)) Disease or
Initiation and Instruction of the Infection)
Specific Immune Response

Humorall R
H Response Acquired
A i d IImmunity,
it Cell-Mediated
C ll M di t d Immune
I
(Extracellular Immunologic Memory, Response
Pathogens and and Protection (Intracellular
Toxins) (Survival) Pathogens and
Viruses)
Factors Influencing Disease Resistance and Immune
Response of Fish
General Specific

Genetics Individuals may exhibit differences in innate

resistance and acquired immunity
E i
Environment
t T
Temperature,
t season, photoperiod
h t i d

Stress Water quality, pollution, density, handling and

transport, breeding
b di cyclesl
Nutrition Feed quality and quantity, nutrient availability,
use of immunostimulants,
immunostimulants antinutritional factors in
feeds
Fish Age, species or strains, individuals

Pathogen Exposure levels, type (parasite, bacterial, viral),

virulence
1From Shoemaker et al.,2001. Immunity and disease resistance in fish. In: Nutrition
and Fish Health (Ed.: Lim, C., Webster, C.D.). Food Products Press, NY. Pgs 149-162.
Immunosuppressant:
pp
Pesticide: Endrin, Malathion, Methyl
b
bromide,Trichlorphon,DDT,
id T i hl h DDT Lindane,
Li d Atrazine
i
Drugs: O
Drugs OTC,
C, Aflatox
Aflatoxin
nBB-1,, Cortisol,
Cort sol, Hydrocortisone
Hydrocort sone

Immunostimulants:
Adjuvant:
j CFA, IFA
Protein: Muramyl dipeptide, Bacterial endotoxins
Drug: Levamisole
Vitamin: Glucans,
Glucans Chitin and chitosan,
chitosan Vit.
Vit C,
C Vit.
Vit E
 Immunosuppressant: Metal
Metal Parameter
Aluminum Reduced chemiluminescence
Arsenic ↑, ↓ phagocytosis
Cadmium ↑ serum antibody in rainbow trout
↓ lymphocyte number and mitogenic response
↓ chemiluminescence
Chromium ↓ serum antibody in brown trout, carp
Copper
pp ↓ serum antibody
y in brown trout
↓ chemiluminescence in rainbow trout
Lead ↓ serum antibody in brown trout
M
Mercury ↓ lymphocyte
l h number
b in b
barb
b
Nickel ↓ serum antibody in brown trout
Zinc ↓ serum antibody in brown trout
Immunosuppressant: aromatic hydrocarbon

Ch i l agents
Chemical t P
Parameter
t
Phenol ↓ antibody-producing cells
Benzidine ↑ nonspecific agglutination
PCB ↓ serum antibogygy
↑ susceptibility to diseases
Chlorinated dioxin Suppressed
pp mitogenic
g response
p
(TCDD)
Polynuclear aromatic ↓ macrophage activity
hydrocarbon (PAHs)
↓ number of
f melanomacrophage
l h