View
226
Download
0
Category
Preview:
Citation preview
STRATEGIES TO SENSE PATHOGENS AND DAMAGED TISSUES
(Bottazzi, Garlanda, Doni, Mantovani Annu Rev Immunol 2010)
TL
R1
-10
LPS, CpG,
Flagellin, Poly I:C... IL-33
-
IL-1
8R
IL-1
RI
IL-1
RII
IL-1
R
AcP
IL-1β IL-1β
IL-1α
- IL-1ras S
T-2
ILRs TLRs
-
TIR domain
Ig domain Leucine - rich repeat
domain MyD88
TIRAP/MAL
TRIF
SARM
TRAM
IL-1
R
AcP
IL-1F9
IL-1F8
IL-1F6
IL-1
Rrp
2
IL-1
8B
P
IL-1
8R
AcP
IL-18
IL-1F7
IL-18
IL-1
R
AcP
IL-1
R
AcP
- Negative
regulation
? Unknown ligand
IL-1
RA
PL
-
IL-1F5
? ? ?
TIG
IRR
-1
TIR
8/
SIG
IRR
-
IL-1F5
-
G
a
r
l
a
n
d
a
(Garlanda, Anders and Mantovani Trends Immunol 2009)
decoy
IL-1
IL-18
IL-33
LPS
CpG
nucleosomes
?? IL-1F5
???
NFkB
IL-1RI
IL-18R
ST2
TIR8/
SIGIRR
TLR4
TLR9
TLR7
??
MyD88
IRAK
Intestinal inflammation
Colitis – associated cancer
TIR domain
Death domain
Conserved aa
(S447, T536)
Replaced aa
(C222, L305)
Pulmonary TB
Autoimmunity
Asthma
Kidney transplantation
Arthritis
Fungal infections
Human:
Chromosome 11, 10 exons
mRNA: 1695 bp
Protein: 410 aa
Mouse:
Chromosome 7, 9 exons
mRNA: 1450 bp
Protein: 409 aa
(Garlanda, Anders and Mantovani Trends Immunol 2009)
Regulation of inflammation by TIR8/SIGIRR
Epithelial mucosal
cells, iDC, Endothelial cells
0 10 20 30 40 50 60 700
25
50
75
100 TIR 8-/-
TIR8 +/+
days
Pe
rce
nt
su
rviv
al
Pulmonary TB- Fungal Inf-
(L.Romani)
Intestinal inflammation and
colitis – associated cancer
Weeks
Perc
en
t S
urv
ival
+/+ -/-
TI
R
D
D
TI
R
TI
R IL-1RI
MyD88
IL-1ra
IL-1
TI
R
-
TI
R
D
D
TI
R
TI
R
MyD88
TI
R
-
LPS
Pro-inflammatory cytokines
Inflammatory chemokines
Leukocyte infiltration
(Garlanda et al PNAS 2005;
Garlanda et al Cancer Res 2007;
Garlanda et al JI 2007;
Lech et al JEM 2008)
Autoimmunity
TIR
8/
SIG
IRR
IL-1
8R
IL-1
8R
AcP
IL-18
IL-1F7
IL-1
RI
IL-1β
IL-1α
IL-1
R
AcP
IL-33
T1/S
T-2
IL-1
R
AcP
TIR
8/
SIG
IRR
TIR
8/
SIG
IRR
Th17 Th2 Th1
- IL-1ra
IL-1
RII
-
-
-
-
-
IL-1
8B
P
-
(Garlanda, Anders and Mantovani Trends Immunol 2009)
IL-8
GCP-2
NAP-2
ENA-78
GROα
GROβ
GROγ
IP-10
Mig
I-TAC
SDF-1α/β
BCA-1
BRAK
MCP-1
MCP-4
MCP-3
MCP-2
MIP-1β
MIP-1αS
MIP-1αP
RANTES
MPIF-1
HCC-1
HCC-2
HCC-4
Eotaxin
Eotaxin-2
Eotaxin-3
TARC
MDC
MIP-3α
ELC
SLC
I-309
TECK
CTACK
PARC
Lymphotactin
SCM-1β
Fractalkine
CXCL8
CXCL6
CXCL7
CXCL5
CXCL1
CXCL2
CXCL3
CXCL10
CXCL9
CXCL11
CXCL12
CXCL13
CXCL16
CXCL14
CCL2
CCL13
CCL7
CCL8
CCL4
CCL3
CCL3LI
CCL5
CCL23
CCL14
CCL15
CCL16
CCL11
CCL24
CCL26
CCL17
CCL22
CCL20
CCL19
CCL21
CCL1
CCL25
CCL27
CCL18
XCL1
XCL2
CX3CL1
CXCR1
CXCR2
CXCR3
CCR4
CCR5
Main targets
CXCR4
CXCR5
CXCR6
Unknown
CCR1
CCR3
CCR6
CCR7
CCR8
CCR9
CCR10
Unknown
XCR1
CX3CR1
CCR2
Chemokine Receptor
Neutrophil
Monocyte
Eosinophil
Basophil
NK cell
B cell
Immature
Mature
mDC
T cell
pDC
Naive
Memory
Th1
Th2
Treg
Inflammatory (RED), homeostatic (GREEN), mixed (YELLOW) (Mantovani, Bonecchi ,Locati Nature Rev Immunol 2006)
Silent vs conventional receptors
Atypical chemokine receptors
No conventional
signaling
(CTX, Ca, MAPK, …)
Signaling chemokine receptors
Signaling
(CTX, Ca, MAPK, …)
The “darc side” of the chemokine system
CXCR7
(Mantovani, Bonecchi and Locati,
Nature Rev Immunol 2006; CGFR 2009)
THE DECOY R D6: A SCAVENGER AND GATEKEEPER OF
INFLAMMATORY CC CHEMOKINE TRANSFER TO LYMPH NODES
Tissue
CCL2
Lymph
Endothelium
CCL2
D6
Lymph node
HEV Blood
(-/- data: Martinez de la Torre et EJI, 2005; PNAS 2007; Jamieson et al Nature Immunol 2005;
Liu et al J Immunol 2006. For review Mantovani, Bonecchi and Locati, Nature Rev Immunol 2006)
M. tuberculosis infection in D6-/-
M. tuberculosis (2 x 103 CFU) intranasal injection
WT (n=10)
D6-/- (n=10)
CFUs
(Di Liberto & Locati et al, J Exp Med 2008)
D6 AS A DECOY AND SCAVENGER: IN VIVO VERITAS
Colitis and colon cancer
Vetrano et al 2010
Mycobacterium tubercolosis
Di Libero Locati et al J Exp Med 2008
Martinez de la Torre Y et al, Eur J Immunol 2005
Granulomatous reaction to CFA injection
Martinez de la Torre Y et al, PNAS 2007
Fetal loss under inflammatory conditions
Jameson Nature Immunol 2005
Nibbs RJ, J Clin Invest 2007
Skin Inflammation and carcinogenesis
Whitehead Am J Res Crit C Med 2007
Otero K et al unpiblished
Allergic airway inflammation
Induction of EAE
(Liu et al JI 2006) Induction of EAE
(Liu et al JI 2006)
Negative regulators (D6; TIR8): 1. tune and balance resistance versus uncontrolled
inflammation; 2. resolution
TIR8 or D6-/-
Death
Inflammation Antimicrobial
resistance
Brakes: TIR8, D6……
Inflammation
TLR/ILR;
chemokines
Equilibrium and
long survival
Antimicrobial
resistance &
MyD88
DECOY RECEPTORS
(updated from Mantovani et al Trends Immunol 2001 )
Caspase/TRAF JAK/STAT
IL-1R TNF IL-10
JAK/STAT G protein
Argos
IL-4/IL13 Chemokines EGF Receptor
family
Core
signaling
Decoys
Decoy RII IL-1
IL-18BP IL-18
Osteoprotegerin RANK, TRAIL
DcR1, DcR2 TRAIL
DcR3 FASL, LIGHT
FDR FASL D6
Inflammatory
CC chemokines
IL-13R2
IL-13
IL-22BP
IL-22
TK
Human Genomic
clone 17 Kb Chr 3q25
Murine gene Chr 3
mRNA 1866 bp
Protein 381 aa
Protomer 43 kDa
Glycosylation 2 kDa Pentraxin domain X domain Signal
peptide
THE PROTOTYPIC LONG PENTRAXIN PTX3
Protein is
mainly
organized in
covalently
linked octamer
Inforzato et al, JBC 2008
(Garlanda et al Annu Rev Immunol 2005, 2010; Bottazzi et al Curr Op Immunol, 2006, 2008)
(Inforzato et al, in revision on J. Biol. Chem.)
Visualization of the rhPTX3 protein by TEM
Shape of the PTX3 protein
Transmission Electron Microscopy (TEM)…
Shape of the PTX3 protein
…and Small Angle X-ray Scattering (SAXS).
(Inforzato et al, in revision on J. Biol. Chem.)
SAXS analysis of rhPTX3 and an ab initio model of the PTX3 protein
"The serum obtained from human
beings and monkeys during the acute
phase of diverse infections contains a
protein which is precipitable by the C
polysaccharide of pneumococcus"
Abernathy and Avery, J. Exp. Med., 1941
Tillet and Francis, J. Exp. Med, 1930
SELECTIVE SUSCEPTIBILITY OF
PTX3 -/- MICE TO A.FUMIGATUS
A.FUMIGATUS CLP L. MONOCYTOGENES
(Garlanda et al Nature 2002)
REGULATION OF PTX3 PRODUCTION IN MYELOMONOCYTIC CELLS
HA MATRIX
DEPOSITION
Mø myeloid
DC
TLR AGONISTS
(e.g. LPS)
PRIMARY INFLAMMATORY
CYTOKINES (IL-1, TNF)
HUMORAL INNATE IMMUNITY
(C; OPSONIZATION VIA FcgR;
NEUTRALIZATION)
IL-10 IFNg - +
TUNING
INFLAMMATION
(Bottazzi et al Annu Rev Immunol 2010)
Modified from Biswas and Mantovani Nature Immunol 2010
The yin-yang of macrophage polarization
M1 M2-like
- intracellular pathogens
- tissue damage
- tumor resistance - parasite encapsulation
- tissue remodeling
- tumor promotion
- immunoregulation
Ligands recognized by PTX3
- Microbial ligands: OMPA, galactomannan and more
- Apoptotic cells
- Complement: C1q, Factor H
- FcgR
- ECM
Outer Membrane Protein A
(Bottazzi et al JBC; Jeannin et al Immunity 2005; Rovere et al Blood 2004; Garlanda et al Nature 2002; Lu et al Nature 2009)
Synergy between Ficolin-2 and PTX3 boosts innate immune recognition
and complement deposition
Ma Y J et al. J. Biol. Chem. 2009, L-ficolin; see also Gout et al J Immunol 2011, M-Ficolin
C4 deposition depends on Ficolin-2 and Ficolin-2/PTX3 collaboration
Ficolin-2/PTX3 cross-talk on A. fumigatus Ficolin-2 isolation from
human plasma by
immobilized PTX3
Classical pathway Antibodies, Pentraxins
Lectin pathway MBL, Ficolins
Alternative pathway C3b-microorganisms
C1q,
C1r,
C1s MASPs
C3b
C4, C2
Factor B,
Factor D,
Properdin C3 and C5
convertases C3a, C5a
C5b-C9
C3b
Inflammation Membrane
attack complex,
lysis
Opsonization
Amplification loop
PENTRAXINS (CRP, SAP, PTX3)
Factor H
C1q
Ficolin
MBL
CRP
SAP
PTX3
Figure 2
(Bottazzi, Garlanda, Doni,
Mantovani, Annu Rev Immunol 2010)
Box 2 | Neutrophil extracellular traps
Neutrophils in the activation and regulation of innate and adaptive immunity
Mantovani A, Cassatella MA, Costantini C, Jaillon S.
Nat Rev Immunol. 2011 Jul 25;11(8):519-31. Review.
Figure 1 | Neutrophils crosstalk with immune and non-immune cells in
inflamed tissues and lymph nodes.
Neutrophils in the activation and regulation of innate and adaptive immunity
Mantovani A, Cassatella MA, Costantini C, Jaillon S.
Nat Rev Immunol. 2011 Jul 25;11(8):519-31. Review.
INNATE IMMUNE RESPONSE
Cellular
innate
immunity
Cytokines,
chemokines,
other effector molecules
C activation,
opsonization,
agglutination
hrs
min TLR
SCAVENGER R
DECTIN-1
CD14
MØ/DC
Microbe, eg
OMPA,
galactomannan
Microbe
COMPLEXITY AND COMPLEMENTARITY OF
CELLULAR AND HUMORAL INNATE IMMUNITY
PMN
Humoral
amplification of
innate immunity
PTX3
(Garlanda et al Nature 2002;
Jeannin, Bottazzi et al
Immunity, 2005; Jaillon et al
J Exp Med 2007)
PAMP
Eg OMPA
JH Lu et al. Nature 000, 1-4 (2008) doi:10.1038/nature07468
Crystal structure of SAP–FccRIIa complex.
PT
X3
CD
11b N
ucle
i
PT
X3
CD
32 N
ucle
i
PT
X3 C
D11b N
ucle
i
PT
X3
CD
11b C
D32
w
PT
X3
w/o
PT
X3
PT
X3 C
D32 N
ucle
i
w PTX3
w/o PTX3
RECRUITMENT OF ACTIVE CD11b AND CD32 IN THE PHAGOCITIC CUP BY PTX3
ROLE OF PTX3 IN FITC-CONIDIA PHAGOCYTOSIS BY ALVEOLAR NEUTROPHILS
A.fumigatus
conidia
FITC-conidia
+
FITC
8X107
i.t.
4h
BAL FITC intensity in leucocytes
(CD45+) and neutrophils
(CD11b+ Ly6G+)
(Moalli et al)
PTX3: LESSONS FROM GENE TARGETING
NON-REDUNDANT FOR:
-FEMALE FERTILITY (ASSEMBLY OF HA-RICH MATRIX; TSG6)
-RESISTANCE AGAINST SELECTED PATHOGENS: AN ANTE-ANTIBODY
(A. fumigatus, P Brasiliensis, P. aeruginosa, K pneumoniae, CMV, Influenza)
-TUNING INFLAMMATION AND IMMUNITY
A UNIQUE, SOLUBLE, MULTIFUNCTIONAL
PATTERN RECOGNITION RECEPTOR
(eg Garlanda et al Nature 2002; Salustri et al Development 2004; Garlanda et al Annu Rev Immunol 2005;
Bottazzi et al Curr Opinion Immunology 2006; Jeannin et al Immunity, 2005; Soares et al Microbes Inf 2006
Romani et al Blood 2006; Reading et al J Immuno 2007; Jaillon et al JEM 2007)
INNATE IMMUNITY - INFLAMMATION
ASN220
Chr 3q25
mRNA 1866 bp
Prot 381 aa
PTX signature
(HxCxS/TWxS)
Ex 1 Ex 2 Ex 3
SP NTD PTX
OmpA Mannans
other
C1q Factor H
FcγR CD11b
? Ficolin-2
Microbe recognition
C activation Opsonization Regulation of inflammation
Repertoire synergism
(Bottazzi et al Annu Rev Immunol 2010)
PTX3 binds P-selectin
(Deban et al., Nat. Immunol. 2010)
PTX3 binds P-selectin via C-terminal (pentraxin)
domain, in a glycosylation-dependent
manner
Kd 360 ± 30 nM (mean of four experiments)
Model of acid-induced acute lung injury (ARDS)
PTX3 attenuates in vivo leukocyte recruitment
(Deban et al., Nat. Immunol. 2010)
In four models of P-selectin-dependent recruitment (pleurisy, ALI/ARDS, thrombin mesenteric inflammation for intravital microscopy, atherosclerosis) and in cancer, genetic deficiency of PTX3 is associated with increased recruitment of PMN or macrophages (TAM)
(Deban et al Nature Immunol 2010 in press; Garlanda et al unpublished data)
Impairment of tethering and rolling on activated endothelium
PTX3
PSGL-1
Psel
Activated endothelium
TLRs TNF-α IL-1
+ IL-10
PTX3
Macrophages
Dendritic cells
Monocytes
PTX3 production in tissues
SYSTEMIC INFLAMMATION
LOCAL INFLAMMATION
PTX3- mediated regulation of leukocyte recruitment
(Deban et al., Nat. Immunol. 2010)
INNATE IMMUNITY - INFLAMMATION
ASN220
Chr 3q25
mRNA 1866 bp
Prot 381 aa
PTX signature
(HxCxS/TWxS)
Ex 1 Ex 2 Ex 3
SP NTD PTX
OmpA Mannans
other
C1q Factor H
FcγR P-selectin Ficolin-2
Microbe recognition
C activation Opsonization Regulation of leukocyte
recruitment and inflammation
Repertoire synergism
(Bottazzi et al Annu Rev Immunol 2010)
PTX3 IN HUMAN PATHOLOGY
• Inflammatory/vascular ( acute myocardial infarction; heart
failure etc; ALI/ARDS; small vessel vasculitis; eclampsia; renal
pathology)
• Infectious (sepsis; meningococcal infection; dengue;
leptospirosis; TB; malaria; monitoring fungal infections in
leukemia)
• Genetic polymorphisms (TB; P. aeruginosa; pregnancy)
Effect of PTX3 in mice with
invasive aspergillosis
Lethally irradiated C3HHeJ mice were
infused with 2x106 T-cell depleted
allogeneic bone marrow cells from BALBc
mice a week before infection with 2x107
Aspergillus conidia. Mice were treated for
5 days before (Pre) or after (Post) the
infection or in concomitance with the
infection.
Dosage:
PTX3 0.04 or 0.2 mg/kg
L-AMB 1 mg/kg
D-AMB 2 mg/kg
PTX3 increases
the therapeutic
effect of L-AMB
and D-AMB
Survival rates and lung fungal
burdens of rats infected with
A. fumigatus and treated with
PTX3 (n = 16).
(A) daily treatment 3 days before
the day of infection and then
for an additional 3 days.
(B) (B) tratment started the day of
infection and then daily for an
additional 3 days.
____ 1.5 mg/kg PTX3;
------- 0.15 mg/kg PTX3; …….
saline.
(C, D) Fungal burden
(galactomannan GMI).
Synergism -
regulation
Activated Mø, PMN, DC
Cellular innate immunity
Cytokines, chemokines,
other effector molecules
days min
epithelia
hr
Microbes-damaged tissues
Cellular sensors: TLR and coreceptors; NLR; RLR; capturing R
DC Mø PMN Endothelium
Humoral, fluid phase, innate immunity
C activation; opsonization; agglutination and
neutralization; regulation of inflammation
PTX3
Properdin
Ficolins
PTX3, C,
M-Ficolin,
SAA3
Pentraxins (CRP, SAP, PTX3)
Collectins (C1q, SP-A, SP-D, MBL)
Ficolins, SAAs
Constitutive and inducible humoral sensors
Figure 6
(Bottazzi, Garlanda, Doni, Mantovani, Annu Rev Immunol 2010)
Antibodies PTX3
Glycosilation-dependent
regulation of inflammation
PTX3 as functional ancestor of antibodies
(ante-antibody)
Agglutination and
neutralization
Opsonization
(FcgR)
Complement
activation
(Bottazzi et al Annu Rev Immunol 2010)
Istituto Clinico Humanitas/ University of Milan Livija Deban Marina Sironi Federica Moalli Sonia Valentino Ivan Cuccovillo Cecilia Garlanda Andrea Doni Barbara Bottazzi Universidade Federal de Minas Gerais, Belo Horizonte Remo Castro Russo Istituto Mario Negri Antonio Bastone Marco Gobbi Vanessa Zambelli Margherita Scanziani Consorzio Mario Negri Sud Gino Evangelista
University of Verona Carlo Laudanna Gabriela Constantin Tecnogen S.p.A Marica Sassano Sigma-Tau Research and Development Giovanni Salvatori Peter Garred Nicole Thielens
Sigma Tau S.p.A. Dr. Giovanni Salvatori
Dr. Ragnar Lindstedt
Istituto clinico “Humanitas”
Università di Milano Prof. Alberto Mantovani
Dr. Barbara Bottazzi
Acknowledgments
Tecnogen S.p.A. Dr. Antonio Verdoliva
Dr. Vincenzo Rivieccio
University of Manchester Prof. Tony Day
All the guys in Day’s group
Dr. Tom Jowitt
Prof. Karl Kadler
Dr. David Holmes
Dr. Clair Baldock
Funding bodies: Federazione Italiana per la Ricerca sul Cancro (FIRC)
Istituto clinico “Humanitas”
Sigma Tau S.p.A.
Tecnogen S.p.A.
Recommended