The role of T cells in the pathogenesis - ankararomatoloji.com fileThe role of T cells in the...
Transcript of The role of T cells in the pathogenesis - ankararomatoloji.com fileThe role of T cells in the...
The role of T cells in the pathogenesis of rheumatic disease
(….. with a focus on RA)
“T cells in health and rheumatic diseases” - Sheraton Hotel, Ankara, 13 March 2009
Andrew P. Cope MD PhDArthritis Research Campaign Professor of Rheumatology
Academic Department of RheumatologyDivision of Immunology, Infection and Inflammatory Diseases
King’s College School of MedicineKing’s College London
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
A working model of disease pathogenesis for RA
progression from UA to RA Healthy
population
little joint destruction
RA
much joint destruction
environmental triggers
genes
immune
response nodisease
auto-antibodies arthritis
onset
diagnosisof RA
non-specific arthritis
spontaneousremission
genotype serotype phenotype
remission
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Susceptible host
Onset of arthritis
Chronicdisease
initiation persistence
tobaccosmoke
trauma
infection
checkpoints defining the evolution and progression of RA
↑ innateimmunity
initiation factors
adaptive i.r. to modified self
antigens
pathways perturbing
immuneregulation
effector cellsmediating
persistence
THERAPY
Checkpoints defining the immunobiology of RA
1
2
pathways regulating
jointintegrity
4
3 5
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Overview of the Lecture
1. What triggers the adaptive immune response in RA?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
What is the molecular basis of adaptive immunity?Thymic selection
a repertoire ofself-reactive
T cells
positive selection
cTECself
ligand
DPThy
TCR
MHCII
CD4/8
Lymphocytes are referential to self-ligands!
Maintenance of T cellsin the periphery
TCR “tickling”of self-reactive
T cells
APCmultipleligands
T cellclonotypicreceptor
dendritic cell
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Immune recognition of “infectious non-self”(as opposed to “non-infectious self”)
DC
T
TCRCD28
MHCIICD80/86Signal 1 “constitutive”Signal 2
“constitutive”
“inducible”
How are costimulatory molecules induced?
pathogen derivedpeptide antigen
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
Lipoproteins(bacteria,
mycoplasma,mycobacteria)
dsRNA(viruses)
LPShsp
Flagellin(bacteria)
ssRNA(viruses) synethetic
compounds
ssRNA CpG DNA Factors fromuropathogenic
bacteria
?Exogenous ligands
Pathogen Associated Molecular Patterns (PAMPs) and Pattern Recognition Receptors (PRRs)
TLR1/6 TLR2 TLR4 TLR5 TLR7 TLR8 TLR9 TLR10 TLR11TLR3
DANGER SIGNALS
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Immune recognition of “infectious non-self”(as opposed to “non-infectious self”)
DC
T
TCRCD28
MHCIICD80/86Signal 1 “constitutive”Signal 2
“constitutive”
“inducible”
How are costimulatory molecules induced?
pathogen derivedpeptide antigen
TLR signal
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
TIR
Lipoproteins(bacteria,
mycoplasma,mycobacteria)
dsRNA(viruses)
LPShsp
Flagellin(bacteria)
ssRNA(viruses) synethetic
compounds
ssRNA CpG DNA Factors fromuropathogenic
bacteria
?Exogenous ligands
Pathogen Associated Molecular Patterns (PAMPs) and Pattern Recognition Receptors (PRRs)
TLR1/6 TLR2 TLR4 TLR5 TLR7 TLR8 TLR9 TLR10 TLR11TLR3
Endogenous ligands
Hsp60,70HMGB1
Gp96HA, HS
FN
chromatin-IgGcomplexes
dsRNA ssRNAssRNA DNAnecroticcells
TLR signal transduction pathways
adapted from O’Neill, 2006
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Inhibition of TLR dependent production of inflammatory mediators
Sacre et al, 2007 Am J Path 170: 518-525
Uninfec
ted
Adβ-gal
AdMyD
88dn
AdMald
n
AdIκBα
0
500
1000
1500
2000
2500
TNFα
(pg/
ml)
Uninfec
ted
Adβ-gal
AdMyD
88dn
AdMald
n
AdIκBα
0
1000
2000
3000
4000
MM
P-3
(ng/
ml)
Uninfec
ted
Adβ-gal
AdMyD
88dn
AdMald
n
AdIκBα
0.0
1.5
3.0
4.5
MM
P-13
(ng/
ml)
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Overview of the Lecture
1. What triggers the adaptive immune response in RA?
2. What do T cells see?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
P4Asp-
β71Lys+
collagen II (1168-1180)N-terminus
C-terminus
Non-associated DRB1*0402β86 Valβ67 Ileβ70 Aspβ71 Glu Pocket 4}
DRα chainα-helix
DRβ chainα-helix
Disease associated DRB1*0401β86 Glyβ67 Leuβ70 Glnβ71 Lys Pocket 4}
Proposed hierarchy of HLA-DRB1 alleles indetermining disease severity in RA
(from Weyand et al, 1995)
*0401/*0401
*0401/*0404
*0401/*01
*0404/*01
*0401/X
*0404/X
*01/*01 nodular vasculitis
*01/X seropositive erosive disease
X/Xnon-erosive erosive
seronegative diseaseANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”
13 Mart 2009 Sheraton Otel, Ankara
HCgp39 in IFA
DRαβ1*0401 DRαβ1*0402
Defining HLA-DR4 restricted T cell responses to cartilage antigens in transgenic mice lacking mouse MHC class II
harvest draining LN @ 10d
evaluate antigen & peptide
specific responses
fuse with TCRneg
thymoma asfusion partner
clone hybridomas
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
DRαβ1*0401 and DRαβ1*0402 present completely distinct sets of HCgp-39 peptide epitopes to CD4+ T cells
0
10
20
30
40
50
60
% a
ll re
spon
ding
hyb
rids
22 28 34 40 46 52 58 64 70 76 82 88 94 100
106
112
118
124
130
136
142
148
154
160
166
172
178
184
190
196
202
208
214
220
226
232
238
244
250
256
262
268
274
280
286
292
298
304
310
316
322
328
334
340
346
352
358
364
368
HCgp-39 peptide
DRαβ 1*0402 (n = 151)
DRαβ1*0401 (n = 250)
Cope et al, 1999 Arthritis Rheum 42:1497-1507
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
The immune response is different (1) a.
0
5000
10000
15000
20000
25000
30000
[3H
] thy
mid
ine
inco
rpor
atio
n (c
pm)
*0401 *0402DRB1*04 subtype
SI - 3.4 SI - 3.6
HCgp-39
medium
0
4000
8000
12000
16000
IFNγ
(pg/
ml)
*0401 *0402DRB1*04 subtype
b.
I-d pool
peptide pool
HCgp-39
Cope et al, 1999 Arthritis Rheum 42:1497-1507
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
0
4000
8000
12000
16000
IFNγ
(pg/
ml)
*0401 (C-line)
*0401(3C4)
*0402
DRB1*04 subtype
0
200
400
600
800
1000
1200
TN
F (p
g/m
l)
*0401 (C-line)
*0401 (3C4)
*0402
DRB1*04 subtype
c. IFNγ d. TNFα
Cope et al, 1999 Arthritis Rheum 42:1497-1507
The immune response is different (2)
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
DC
Teff
TCRCD28
MHC class II
B disease specificautoantibodies
antigen experiencedeffector T cells
Defining the key autoantigenic determinants
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
What have RA-specific autoantibodies taught us?
reactivity to“modified”
self
“stress”
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
reactivity to“modified”
self
“stress”
citrullinatedproteins
arginine → citrulline“neoepitopes”
What have RA-specific autoantibodies taught us?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
reactivity to“modified”
self
“stress”
inflammation
infiltration
PAD2/4
citrullinatedproteins
mΦ, PMN
vimentin, fibrin, histones, α-enolase
deiminationCa2+
Schellekens, Vossenaar, Tak, van Venrooij et alinflammation specific
What have RA-specific autoantibodies taught us?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
reactivity to“modified”
self
“stress”
inflammation
infiltration
PAD2/4
citrullinatedproteins
mΦ, PMN
PAD4 geneticpolymorphism
↑ mRNAstability
deiminationCa2+
Schellekens, Vossenaar, Tak, van Venrooij et alinflammation specific
Suzuki et al, Nat Gen 2003
What have RA-specific autoantibodies taught us?
vimentin, fibrin, histones, α-enolase
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
reactivity to“modified”
self
“stress”
inflammation
infiltration
PAD2/4
citrullinatedproteins
HLA class II
T cellresponse
B cellresponse
mΦ, PMN
PAD4 geneticpolymorphism
↑ mRNAstability
synovialplasma cells
anti-CCP
TH
IgG1
inflammation specific disease specific
deiminationCa2+
Suzuki et al, Nat Gen 2003
?
What have RA-specific autoantibodies taught us?
vimentin, fibrin, histones, α-enolase
P4Asp-
β71Lys+
collagen II (1168-1180)N-terminus
C-terminus
Non-associated DRB1*0402β86 Valβ67 Ileβ70 Aspβ71 Glu Pocket 4}
DRα chainα-helix
DRβ chainα-helix
Disease associated DRB1*0401β86 Glyβ67 Leuβ70 Glnβ71 Lys Pocket 4}
Arg not favouredCit permissiveat P4
Overview of the Lecture
1. What triggers the adaptive immune response in RA?
2. What do T cells see?
3. Have any immunogenetic studies been informative?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
DC
Teff
TCRCD28
modified“signal output”
Functional impact
Thymic selection
Cell survival
Th differentiation
Migration
Effector responses
Immune regulation
Gene polymorphism
HLA-DRB1
CIITA
PADI4
PTPN22
PD1
CTLA4
CD25
STAT4
“signal input”
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Overview of the Lecture
1. What triggers the adaptive immune response in RA?
2. What do T cells see?
3. Have any immunogenetic studies been informative?
4. What do RA T cells look like?
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Prediction:
1.Activated phenotype2.Features of prior antigen exposure3.Extensive proliferative activity4.Expansions of selected (high affinity) clones5.Reactive to tissue specific (cartilage, synovium, bone)
antigens6. Features of distinct Th lineage differentiation7. Intrinsic survival advantage8. Potent migratory capacity9. Impaired regulatory function
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
RAsynovial
T cell
RAsynovial
T cell
senescence
memory effector
migration
antigen recognition
effectorresponse
TCR
TCRζdim
costimulation CD28
OX40
ICOSL
CD28null
CD45RO,CD45RBdim
CD44,CD69,CD62Llo
CCR7lo
VLA-1,VLA-4
LFA-1
CCR4,CCR5,CCR6, CXCR3,CXCR4,CXCR5
NKG2D
CX3CR1
CD25neg
CD27neg
CD40Lneg
IL-4RTNF+
IFNγ+
IL-17+
IL-10+
RANKL
H202+
mTNF
LTα1β2
IL-6R
IL-23R
IL-15R
TNFR
Cope AP 2008 Arthritis Res Ther
The reality:
Bcl-2lo
Bcl-XLlo
Overview of the Lecture
1. What triggers the adaptive immune response in RA?
2. What do T cells see?
3. Have any immunogenetic studies been informative?
4. What do RA T cells look like?
5. Multiple pathways of T cell effector function
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
The TCR/CD3 antigen receptor complex
TCR/CD3 complex
TCRζbright
TCRζdim
CD3εTC
Rζ
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
TCRζdim cells are enriched for effector Th cell subsets
Zhang et al, Blood 2007
Data.027
100 101 102 103 104SSC-WIL-17
TCR
ζ
CD4+
TCRζdim
%IF
Nγ
expr
essi
ng c
ells
TCRζdimTCRζbright
P < 0.0005
0
10
20
30
40
50
60
70(A) (B)
(stim: PMA and ionomycin)
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
Regulatory CD4+ T cell subsets reside in the TCRζbright population%
IL-1
0 ex
pres
sing
cel
ls
TCRζdimTCRζbright
P < 0.0005
Zhang et al, Blood 2007
(A)
0
10
20
30
40
50
0
10
20
30
40
50
60
70
80
90
100
%Fo
xp3+
cells
(B)
TCRζdimTCRζbright
P < 0.0001
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
healthy donorPBL
transwell
EC
gelatin
TCRζbright
TCRζdim
TNF
Migration of TCRζdim T cells across activated endotheliumlower chamberupper chamber
CD3
TCR
ζ
88.5%
11.5%
25%
75%
bright dim bright dim bright dim0
20
40
60
80
100
% c
ells
mig
ratin
g
TCRζ expressing subset CD3 CD4 CD8
p < 0.008 p < 0.035 p > 0.16
Zhang et al, Blood 2007ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”
13 Mart 2009 Sheraton Otel, Ankara
- +--
- - - - -- - - - -
+ ++
+- - -
+
--0
500
1000
1500 IFNγ IL-10
Accessory signals are intact in TCRζdim T cells
in collaboration with Fionula Brennan TCRζdim cells generated by stimulation with IL-2, IL-6 and TNFα
cyto
kine
pro
duct
ion
(pg/
ml)
TCRζdim cells plus: CHO/vector:CHO/CD80:CHO/CD86:
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
0
50
100
150
200
250
300m
onoc
ytes 3:
1
5:1
7:1
T cells:monocytes
TNFα
(pg/
ml)
(TCRζdim)(TCRζbright) mon
ocyt
es
T ce
lls
mon
ocyt
es+
LPS 3:1
5:1
7:10
100
200
300
4004000
5000
T cells:monocytes
ζbright ζdim
Analysis of contact dependent effector responses
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
0
10
20
30
40
50
60
70
80
CD3ε
TCR
ζ
TCRζdim T cells are enriched at sites of inflammation
SF
PB SF SM
CD3+ T cells
% T
CR
ζdim
T ce
lls
PB
Zhang et al, Blood 2007ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”
13 Mart 2009 Sheraton Otel, Ankara
% C
D3+ T
CR
ζdim
cells
PB SF SM 0
10
20
30
40
50
60
70
80
TNF blockade
effector T cells
TNF blockade
• memory, activated• CCR/CXCR profile• integrins• senescence• Ag engagement
PHENOTYPE
Persistence of TCRζdim effector T cells after infliximab therapy
circulating TCRζdim
synovial tissue
high disease activity
circulating TCRζdim
synovial tissue
low disease activity
CD
3TCRζ
11.1% 88.9%
TCRζ
45.4% 54.6%
KEY MESSAGES
1. The phenotype and function of PB and synovial T cells points to a key role for inflammation and aging in the pathogenic process.
2. Expression of costimulatory molecules can be exploited for therapeutic process.
3. Antigen determinants for T cells remain poorly define. Extendedautoantibody profiling is likely to provide insights in the future.
4. T cell effector pathways are complex. The precise role of Th1, Th2 and Th17 cells requires further investigation.
5. Deeper understanding of the basis for defective immune regulation is urgently required.
6. Genome wide association studies have provided new clues to the perturbations of adaptive immunity in chronic inflammatory autoimmune diseases such as RA.
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara
FLS
ACUTE PHASIC CHRONIC counter-
regulatorynetworks
TLR inhibitors IL-4, IL-10, TGFβ, IL-13, L-11, IL-2 IL-1Ra, sTNF-R, IL-10, IL-18bp, OPG, adiponectin
transmigration of monocytesPMN, mast cells and NK cells
tissue damage,TLR ligands
TLR
IFNs +survivalfactors
1, TNF, IL-6, IL-22PGE2, bFGF, PDGFEGF, VEGF, TGFβ
COX, LOX
CXCL5/8/9/10/12/13,CCL2/3/5/20/21
CX3CL1
LTβR
activation of endothelium,↑ adhesion molecules
INITIATION ANTIGEN MODE INFLAMMATION MODE
B
Teff
MΦ FLS
Adipo
TCRζdimCD28null
Treg
IL-2deficiency
+ IL-6
IL-1, IL-6, TNFIL-15, IL-32GM-CSFoncostatin MM-CSF, VEGFChemokinesadipokinesHMGB1IL-10
tissue response
adipokines
osteoclasts chrondrocyte
TNFM-CSF
OPG
RANKL
IL-17+
MMPADAMADAM
TNFIL-1
onco-statin M
PGE2
features of inflammation and repair
DC
Teff
TCR
CD28
B
Treg
CD40L
CD40
IL-2
EARLY LATE IL-2, IL-4, IFNγIL-13, TGFβ IL-17IL-15 RANTESGM-CSF TNF
LTα/βIL-10
IL-12, IL-23IL-15, IL-18
IL-6IL-10chemo-kines
AutoAb
BLySAPRIL
TGFβIL-10
TGFβIL-10
CD80/86
Credits ………
TCRζ genetics
Claire Gorman
Andrew RussellTim Vyse
mRNA stability
Andy Clark (3’UTR)
TCR signalling
Joanna ClarkZhuoli ZhangPia IsomäkiKarolina AleksiyadisNina Panesar
Federica Marelli-BergClaudia MonacoEnrico Ammirati
Alex AnnenkovYuti Chernajovsky
SKG mouse studies
Xiang WuOli SomenziTharsana Tharmalingam
Shimon Sakaguchi
FundingWellcome TrustarcMRCEU
ANKARA ROMATOLOJİ SEMPOZYUMU “Sağlıkta ve Romatizmal Hastalıklarda T Hücreleri”13 Mart 2009 Sheraton Otel, Ankara