Post on 13-Jun-2015
Department of Immunology
Fabienne Mackay
Immunology outside the square:Are immune functions influenced by otherbiological systems in the body such as the
neurological functions?
IMM3042
Developing Immunology of the future
Thinking outside of the box
The immune system
1- CNS
2- endocrine system
3- metabolism
4-TraditionalChinese medicine
5- Epigenetics (mirRNA,methylation)
6-cancers
Fabienne MackayDepartment of Immunology
Psychological stress and immunedefences
The adaptive immune response Antigen
APC such as a DC
T cells
AgLPS
“Mature” DC
RecirculationThrough lymphoid tissue
Th1Th2
•Cell-mediated immunity•Defense againstintracellular pathogens
•Allergy•Defense againstextracellular pathogens
BCD4
T
B cell follicleT cell area
Effector cells
BTc
The T helper paradigm
Th1cell
IL-12
IFNγ
IFNγ
NaïveCD4+
T cell
Autoimmunity DTH responses
Th2 cell
IL-4
IL-4
Allergy, asthma IL-4
Th17cell
TGF-βIL-23, IL-6
AutoimmunityInflammationIL-17
IL-12 and the regulation of Th1/Th2
T cellIL-12
IFNγGM-CSFTNFIL-8 Proliferation
Th2
Macrophages, neutrophils, DCs
Th1
B cell
IgE IgA
IFNγ IgG2a
An example of interaction between the neuronal and immunesystems
A fundamental bimodal role for neuropeptide Y1 receptor in the immune system and amodulator of cytokine production.
Stress
Neuropeptide Y
Immune cells
Neuropeptide Y
• Present in the peripheral, central and sympatheticnervous systems• Also been found in nonsympathetic neurons in severalorgans• Highly conserved• Family of hormones with peptide YY (PYY) andpancreatic polypeptide (PP)
NPY mRNA distribution in themouse brain
NPY
PYY
PP
Y P S K PD N P G E D A P A E D MA R Y Y S A L RH Y I N L I T RQ R Y
Y P I K P E A P G E D AS P E E L N R Y Y AS L RH Y L N L V T RQ R Y
A P L E P V Y P G D N A T P E QMAQ Y A AD L R R Y I N M L T R P R Y
Neuropeptide Y receptors
• 5 receptors: Y1, Y2, Y4, Y5, y6
• 7 transmembrane GPCRs of Rhodopsin-like superfamily
• NPY and PYY identical affinity for all Y receptors, PP prefers Y4
• Act in an inhibitory fashion, eg inhibition of cAMP accumulation
• NPY induces proliferation via Y1 / MAPK pathway
Seizure Protection
Heart RateNPY
Pain
Nerve Regeneration
Memory
Anxiety
Bone Homeostasis
Thermogenesis
Alcohol ToleranceMetabolic Rate
Blood Pressure
Aggression
Reproduction
Feeding Behavior
Circadian Rhythm
Immune System
Expression of NPY receptor Y1 by immune cellsY1 receptor expressionin leukocytes
Y1Rβ-tubulin
Human leukocytes, cell-sorted
brainCD4 CD8 NK B Mφ PMN
Y1R18S
Rat PBMCY1RGAPDH
Mouse lymph-node cells
brainLN T
Bedoui et al.
Mouse leukocytes, cell-sorted
+ spl B CD4 CD8 Mφ DC NK MC Y1-/-
MφWheway, 2005
Y1R
Phenotype of Y1-/- mice
⇔ Food intake
⇑ Body weight
⇑ Adiposity
⇑ Nociception (perception of pain)
⇑ Aggressive behaviour
⇑ Alcohol consumption
⇔Heart rate
⇑Fertility
⇑Neurogenesis
Immune responses?
Metabolic
Behavioural
Neurological
Physiological
Immune phenotype of Y1-/- mice
***
Basal serum antibody
0
5
10
15
Y1
WT
IgG1 IgG2a IgG2b IgG3 IgA IgM
Titre
(log
2)
**
0
5
10
15
Y1
WTWT Y1-/-
0
5
10
15
Y1
WT
• Smaller spleens•⇓ mature B cells in spleen,
mesenteric and peripheral LN andperitoneal lavage•Lymphocyte survival normal invitro
•T cell development in thymus
normal
JW281003PLN.020JW281003PLN.020
100
101
102
103
104
CD44
100
101
102
103
104
CD44
JW281003PLN.024JW281003PLN.024
100
101
102
103
104
CD44
100
101
102
103
104
CD44
JW281003PLN.020JW281003PLN.020
100
101
102
103
104
CD44
100
101
102
103
104
CD44
JW281003PLN.024JW281003PLN.024
100
101
102
103
104
CD44
100
101
102
103
104
CD44
0
500
1000
1500
CD4 CD8
Y1-/-
WT
0
50
100
150
200
CD4 CD8
Y1-/-
WT
Altered T cell phenotype in Y1-/- mice
Naive
Effector
Y1+/+ Y1-/-
CD44
CD62
L
CD4
CD8
51.5
12.4
80.8
3.5
23.6
10.0
65.5
2.6**
*
*
*
Y1+/+
Y1-/-
Y1+/+
Y1-/-
N.B. Percentage of total CD4 or CD8 shown
cells
(x10
3 )ce
lls (x
10 3 )
The Delayed-Type Hypersensitivity (DTH) protocol
• Classic T helper 1 (Th1) mediated model
•Day 0 •Day 7
Sensitisation(mBSA/CFA)
Footpad re-challenge(mBSA)
•Day 8 •Day 9
Measurement offootpad swelling
• Experimental protocol:
•Day 14
Culled for serumantibody ELISA
0
25
50
75
100
Column 5
Column 4
Y1
WT
Reduced Th1 responses in Y1-/- mice
0
2500
5000
7500
10000
Y1-/-
WT
Column 7
WT
DTH footpad swelling
WT Y1-/-
WT
Y1-/-
**
IL-4IFN-γ
WT Y1-/- WT
pg/m
l cyt
okin
e
*
Y1-/-
% in
crea
se in
sw
ellin
g
Conflicting observations
T cells T cells?Y1-/-Y1+/+
NPYOr Y1 agonist
Inhibition of Th1 responsesInhibition of EAE
Inhibition of Th1 responses
-How can we reconcile the idea that Y1 is a negative regulator of Th1 T cellfunction with the fact that Y1-/- mice are protected against Th1-mediateddisorders?
Others have shown:Bedoui et al. 2004
JI, Trends Immunol.Vol 10, pp 508-512
We are showing this:
A possible explanation
Antigen
APC such as a DC
T cells
AgLPS
“Mature” DC
RecirculationThrough lymphoid tissue
Th1Th2
•Cell-mediated immunity•Defense againstintracellular pathogens
•Allergy•Defense againstextracellular pathogens
BCD4
T
B cell follicleT cell area
Effector cells
BTc
Y1 signalling
+-
Step 1Step 2
Using Y1 agonists
Antigen
APC such as a DC
T cells
AgLPS
“Mature” DC
RecirculationThrough lymphoid tissue
Th1Th2
•Cell-mediated immunity•Defense againstintracellular pathogens
•Allergy•Defense againstextracellular pathogens
BCD4
T
B cell follicleT cell area
Effector cells
BTc
Y1 signalling
+-
Step 1Step 2
No Th1
Using Y1-/- mice
Antigen
APC such as a DC
T cells
AgLPS
“Mature” DC
RecirculationThrough lymphoid tissue
Th1Th2
•Cell-mediated immunity•Defense againstintracellular pathogens
•Allergy•Defense againstextracellular pathogens
BCD4
T
B cell follicleT cell area
Effector cells
BTc
Y1 signalling
+-
Step 1Step 2
No Th1
To prove this we had to test Y1-/- T cells and APCseparately
T cell responses in Y1-/- mice
0
50
100
150
200
250
0 2.5 5 7.5 10 12.5
11/4/05 aCD3 proliferation exp
Y1
WT
[3HT
]thym
idin
e up
take
(cpm
x10
3)
• Y1-/- T cells are hyper responsive to αCD3 stimulation and undergo more divisionin vitro
αCD3 response
αCD3 (µg/ml)
*
***
WT
Y1-/-
Cell division
WT
Y1-/-
CFSEce
ll nu
mbe
r
Model of autoimmune colitis
•Rag1-/- mice are recipients – noT or B cells•Naïve CD4 T cells(CD4+CD45RBhi) are donor cells•In the lymphopenic environment,T cells proliferate and areactivated by an unknown gutantigen resulting in colitis•Mice develop colitis after 21-28days
CD4+CD45RBhi transfermodel
Y1-/- T cells hyperproliferate in lymphopenichost resulting in severe colitis
-20
-15
-10
-5
0
5
0 10 20 30
Data out to day 28
Column 9
Column 7
% w
eigh
t cha
nge
Days
***
WT donor Y1-/- donor
CD4+ CD45RBhi transfer colitis model in RAG1-/- mice
***
**
0
250
500
750
1000
IFNg
IFNg Y1
IFNg WT
0
25
50
75
100
CD4 T
CD4 Y1
CD4 T
pg/m
l IFN
-γ
CD4+
in s
plee
n (x
10 3 )
*****
WT donor Y1-/- donor WT donor Y1-/- donor
F
Y1+/+ T cells ⇒ RAG-/-
Y1-/- T cells ⇒ RAG-/-
Y1+/+ T cells ⇒ RAG-/-
Day12
Day28
What about the function ofY1-/- APCs ?
•Y1-/- T cells are hyper-responsive and can differentiate into Th1 effector cells in the
presence of WT APCs
•Y1 receptor is a critical negative regulator of T cell activation
Reduction of DTH in Y1-/- mice is not due to an intrinsic T cell defect
100
101
102
103
104
OVA
Y1-/- DCs are functionally impaired
FITC- OVA
WT
0
500
1000
1500
2000
2500
Y1-/'-
WT
WT Y1-/-
% in
crea
se in
MFI
**
•Bone marrow derived DC (BMDDC) generated in presence of GM-CSF & IL-4
Y1-/-
Antigen uptake in vitro
[3HT
]thym
idin
e up
take
(cpm
x10
3)
0
100
200
300
stim only iDC LPS aCD40
Y1
WTWT
Y1-/-
***
***
anti-CD40
LPSiDCstim only
Allogenic MLR- Y1-/- DCsas stimulators
Y1-/- DCs are functionally impaired in vivo
0
25
50
75
100
DC DTH model data
Column 15
Column 14
WT/Y1 DC
WT/WT DC
DC induced antigen-specificT cell priming in vivo
% in
crea
se in
foot
pad
swel
ling
*
WTBMDDC
Y1-/-
BMDDC
Pulsed withmBSA
DC
Day 7Foot pad injection of mBSA
Swelling
-5
0
5
10
0 3 6 9 12
Days
RAG_Y1 RB colitis model
Y1 RAG
WT RAG
Rbhi colitis model in RAG1-/-Y1-/- mice
*
Y1-/- DCs are functionally impaired
WT/RAG
Y1-/-/RAG
% w
eigh
t cha
nge
Defective IL-12 production in Y1-/- mice
0
500
1000
1500
2000
2500
Column 5
Column 4
Y1-/-
WT
Homeostatic serumconcentration of IL-12
pg/m
l IL-
12 **
WT Y1-/-
resp WT 0
5
10
15
24 48 72
Time
DC prep# 7
Y1 LPS
WT LPS
Y1 con
WT con
0
5
10
15
20
25
24 48 72
Time
DC prep# 7
Y1 CD40
WT CD40
Y1 con
WT con
ng/m
l IL
-12
LPSanti-CD40
IL-12 production in stimulatedBMDDCs
WT Y1-/-
WT Y1-/-
Lethal irradiation
WT Y1-/-Inject BM cells fromdonor mouse
WT⇒Y1-/- and Y1-/-⇒WT
Y1-/- BMWT BM
WT Y1-/-
6 weeks reconstitution
DTH model
Chimeras
WT mouse withY1-/- ImmuneSystem
Y1-/- mouse withWT ImmuneSystem
Inhibition in Th1 models is immunecell intrinsic
0
20
40
60
80
100
Column 33
Column 32
Column 31
Column 30
Y1 (WT BM)
WT(Y1 BM)
Y1-/-
WT
Y1-/-
WT
WTY1-
/-
**
% in
crea
se in
sw
ellin
g
*
DTH
Y1-/-
WT
T cells
Y1 signalling
APC such as a DC
+-
Bimodal functions of Y1 receptor
-How this system fits with other known immuno-regulatory systems?
-What happens when exogenous levels of NPY rise ( e.g. stress)
Wheway et al. 2005, J. Exp. Med., 202:1527-1538
Bimodal functions of Y1 receptor
T cellAPC
Step 2Step 1
Y1
Activation
PresentationY1 agonist
OK
Y1
Stop
Y1 agonist
Y1
Y1-/- mice APC
Y1
T cell
StopStop
Activation
Presentation
Le Monde, FranceDecember 2005
The Australian, Dec 5th 2005 The Age, Dec 5th 2005
The Telegraph, Dec 5th 2005
Relevance in the clinic
-Y1 expression is reduced on T cells from patients with schizophrenia
-Well documented defects in Th1 responses inpatients with schizophrenia
-Schizophrenic patients have reduced numbersof circulating activated T cells
-T cells from schizophrenic patients are hyper-responsive ex-vivo but these patients havereduced in vivo T cell responses to tuberculin
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