innate and adaptive immunity- complementary & distinct ... · innate and adaptive...
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Innate and Adaptive Immunity-Complementary & Distinct Host Defenses
Innate and Adaptive Immunity-Complementary & Distinct Host Defenses
INNATE ADAPTIVE
Hard-wired Requires Priming
Immediate Delayed or Immediate
Cell bound receptors Soluble and Cellular
Pattern-molecule recognition Antigen Specific
Macrophages, NK, non-prof. APC B-cells, T-cells
Different ligands selectively activate different TLRs
Different ligands selectively Different ligands selectively activate different TLRs activate different TLRs
TLR
2TL
R2
TLR
4TL
R4
TLR
5TL
R5
TLR
6TL
R6
TLR
9TL
R9
TLR
3TL
R3
TLR
1TL
R1
TLR
7TL
R7
TLR
8TL
R8
TLR
11TL
R11
FlagellinFlagellin
LPS LPS
CpG DNACpG DNALipoproteinsLipoproteins
ds RNAds RNAss RNAss RNA ProfilinProfilin--likelike
ProtozoanProtozoan
TLRs are expressed by many cell types throughout the gastrointestinal tract
TLRs are expressed by many cell types throughout the gastrointestinal tract
IEC (IEC (stomachstomach, SI, , SI, coloncolon))MonocytesMonocytes//macrophagesmacrophagesDendriticDendritic cellscellsMyofibroblastsMyofibroblastsEndothelialEndothelial cellscellsAdipocytesAdipocytes
TLR4TLR4
CarioCario et al. Am J et al. Am J PatholPathol 2002 160:1652002 160:165
IECIEC
apicalapical
In constrast, TLR5 is preferentially expressedat the basolateral pole of differentiated IEC
In constrast, TLR5 is preferentially expressedat the basolateral pole of differentiated IEC
SubmucosaSubmucosa
LumenLumen
J. Immunol. 2001 167:1882J. Immunol. 2001 167:1882
Infect. Immun. 2000 68:7010Infect. Immun. 2000 68:7010
T84T84
ColonColontissuetissue
Cell
Nucleus
TLRs induce multiple signal transduction pathwaysvia different adaptor proteins
TLRs induce multiple signal transduction pathwaysvia different adaptor proteins
resident resident microfloramicroflora
Mechanisms of intestinal epithelial ”TOLLerance”
Mechanisms of intestinal epithelial ”TOLLerance”
Decreased Decreased surface expressionsurface expression
IntestinalIntestinalEpitheliumEpithelium
NFNFκκBB
Ligand neutralizationLigand neutralization
Mucins DefensinsMucins DefensinsIgA TrefoilIgA Trefoil
Receptor localizationReceptor localizationState of differentiationState of differentiation
TLR4TLR4
NormalNormal
ST2ST2 TollipTollip
Signaling suppressorsSignaling suppressors
PPARPPARγγ
TGFTGFßß SOCSSOCS
TRIADTRIAD3A3A
CytokineCytokine
Intestinal epithelial TLR4 expression is significantly increased in IBD
Intestinal epithelial TLR4 expression is significantly increased in IBD
HealthyHealthy
CrohnCrohn’’ss DiseaseDisease
Ulcerative ColitisUlcerative ColitisCario, Podolsky. Infect Immun 2000;68:7010Cario, Podolsky. Infect Immun 2000;68:7010
TLR4 alteration associated with IBDTLR4 alteration associated with IBD
Upregulation of mucosal TLR4 expression in active, human IBD
Association of TLR4D299G-polymorphism with IBD in selective populations
Murine TLR4-“gain-of-function”: spontaneous intestinal inflammation with TH1-excess
Murine TLR4-“loss-of-function”: increased susceptibility to DSS-colitis and gram─ infections
Flagellins stimulate aberrant intestinal immune reactions via TLR5 as dominant antigens in colitis Flagellins stimulate aberrant intestinal immune
reactions via TLR5 as dominant antigens in colitis
Lodes et al. JCI 2004;113:1296Lodes et al. JCI 2004;113:1296
••Marked Marked TT--cellcell reactivityreactivity againstagainst flagellinflagellin••FlagellinFlagellin--specificspecific T T cellscells induceinduce colitiscolitis in SCID in SCID micemice••SerologicalSerological responseresponse to to specificspecific flagellinsflagellins in CD in CD ptspts..
TLR4 mutant and knockout mice are highly susceptible to develop DSS colitis
TLR4 mutant and knockout mice are highly susceptible to develop DSS colitis
Fukuta et al. Am J Physiol GI 2005;288: G1055
4.154.15±±0.790.79C57BL6/JC57BL6/J
7.187.18±±0.560.56C3H/HeJC3H/HeJ
DSS colitis scoreDSS colitis scoreMouse strainMouse strain
Mahler et al. Am J Physiol GI 1998;274: G544
RakoffRakoff--Nahoum et al. Nahoum et al. Cell 2004;118:229Cell 2004;118:229
DSS 2%, 7 days, F2DSS 2%, 7 days, F2
Basal TLR activation may be protective against acute injury through its beneficial effects in the intestinal epithelium
Basal TLR activation may be protective against acute injury through its beneficial effects in the intestinal epithelium
EpitheliumEpithelium
Acute injuryAcute injuryintestinal epithelial barrierintestinal epithelial barrier
Com
men
sals
Com
men
sals
Healing ↑Barrier protection ↑Tolerance ↑
TLR
TLR2 assists the host to confer intestinal epithelial barrier function
TLR2 assists the host to confer intestinal epithelial barrier function
0
50
100
150
200
0 15 30 45 60 75 90 105 120
Time (mins)
TER
(% o
f neg
ativ
e co
ntro
ls)
TLR2DN TLR4DN
+ PCSK* *
# #
°°
TLR4DNTLR4DN-- vs. TLR2DNvs. TLR2DN--transfected monolayers: transfected monolayers: *p<0.01; #p<0.001; *p<0.01; #p<0.001; °°p<0.05p<0.05
Cario et al. Gastroenterology 2004;127:224Cario et al. Gastroenterology 2004;127:224
XX--YY
ZOZO--11-- PC
SKPC
SK
ZZ
+ PC
SK+
PCSK
XX--YY
ZOZO--11
ZZ
IECIEC
Commensal compounds assist the host to confer intestinal epithelial barrier functionCommensal compounds assist the host to confer intestinal epithelial barrier function
Madsen, et al. Madsen, et al. Gastroenterology 2001;121:580Gastroenterology 2001;121:580
T84
Otte, Podolsky. Am J Physiol GastrointestOtte, Podolsky. Am J Physiol GastrointestLiver Physiol 2004;286:G613Liver Physiol 2004;286:G613
+LPS 10ng/ml+LPS 10ng/ml MMθθ CD4+ T cellsCD4+ T cells
controlcontrol TLR4TLR4--//-- TLR4/Stat3TLR4/Stat3--//--
in thein the absence of absence of TLR4, the onset of TLR4, the onset of chronic chronic enterocolitisenterocolitisis delayed,is delayed, and and activity of colitis is activity of colitis is improvedimproved
Kobayashi et al. JCI 2003;111:1297Kobayashi et al. JCI 2003;111:1297
Commensal-mediated TLR4 dysfunction contributes to the development of TH1-intestinal inflammation
Commensal-mediated TLR4 dysfunction contributes to the development of TH1-intestinal inflammation
Com
men
sals
Com
men
sals M
ucosaM
ucosaInvasion
TLRTH1TH1--typetype
CytokinesCytokines
Aberrant TLR4/MyD88 Aberrant TLR4/MyD88 signalingsignaling may may favorfavor chronicchronicintestinal inflammation through intestinal inflammation through commensalcommensal--mediated mediated
proinflammatoryproinflammatory TH1TH1--effects in the lamina effects in the lamina propriapropria
Injury ↑Barrier destruction ↑Intolerance ↑Chronic inflammatory diseaseChronic inflammatory disease
Therapeutic Exploitation of the InnateImmune System
Activation?
Inhibition?
Innate Immune Responses are Mediated by Several Receptor Classes
• TollToll--like receptorslike receptors
• NodsNods
• Cd1
• Other
Mutations in LRRs of NOD2 link tothe risk of Crohn’s disease
Gene
Protein
NBD LRRCARD1 1040
R702W
G908R
3020insC
0
20
40
60
80
1 0 0
1 2 0
1 4 0
1 2 3 4
100
80
60
40
20
0
120
140CFU % vs Caco2
***
***
p<0.05 vs Caco2
p<0.05 vs MOCK
CD-mutant Nod2 exhibits defective epithelial killingof intracellular invasive bacteria
NOD2 is membrane associated
- - E A L L Q A L E R N D T I L E V W L R G N T F S L E E V D K L G C R D T R L L L
NOD2insC3020
Amino acid sequences of the C-terminal domain of NOD2 and NOD2 3020insC mutant associated with Crohn’s disease.
IDENTIFICATION OF GRIM-19 AS NOD2 INTERACTING PROTEIN
COS7
/MO
CK+
Xp-G
RIM
19
COS7
/Fla
g-NO
D2+
Xp-G
RIM
19
GRIM19
NOD2
GRIM19
IP anti-NOD2WBanti-GRIM19
anti-NOD2
WBanti-GRIM19(anti Xpress Ab)
1. Immunoprecipitation 2. Immunostaining
Merge
XpGRIM19
GFP-NOD2
GRIM-19 ACTS DOWNSTREAM OF NOD2 AND IS REQUIRED FOR NF-kB ACTIVATION
0
20
40
60
80
100
120
- MDP
+ MDP-LD
NOD2 1 nggrim-19 siRNA-1control grim-19 siRNApSUPERgrim-19 mRNA level
----1
+---ND
++--0.32*
+--+0.98
*
p=0.027
Rel
ativ
e lu
cife
rase
act
ivity
+-+-0.89
p=0.218
Crohn’s Disease is Associated with Reduced Early Inflammatory Response to Subcutaneous Injection of E coli
Crohn’s Disease is Associated with Reduced Early Inflammatory Response to Subcutaneous Injection of E coli
Marks et al, Lancet, 2006
Healthy ControlHealthy Control
CrohnCrohn’’s s DiseaseDisease
Nod2/Card 15 Reduce TLR-2 mediated IL-12 and IFN Production
Nod2/Card 15 Reduce TLR-2 mediated IL-12 and IFN Production
Watanabe et al, Nature Immun, 2004
Multiple Stimuli Converge on a Common Signaling Pathway in IBD
MDPNod 2
LPS/PGN
TLR 4/TLR2
Myd88
NIK
IKK
NF-κB
ILs/CC
Apoptosis?
TNF/TNFR
IL1/IL1R
OCTN1?DLG5?
Nicolas Barnich Hans Christian ReineckerElke Cario Ramnik XavierTakakazu HisamatsuJan-Michel Otte Beth McCormickManabu SuzukiTomo TeradaJesus Yamamoto-FurushoJose AguirreKathryn Devaney