Role of Innate Immunity and Inflammation
Role of Innate Immunity and Inflammation
Cecilia Garlanda Istituto Clinico Humanitas Milan, Italy Activation of Innate Immune responses and Inflammation The inflammatory response The humoral and the cellular arms of innate immunity
Ficolins Collectins Biological (MBL, SP-A, SP-D) fluids C activation C1q
Short and long opsonization Pentraxins (CRP, SAP, PTX3)
Capturing R FcRC-type lectin SR integrin Lipopeptides, OmpA, LPS, PGN
MDP TIR TIR NALP3 (inflammasome) TLR1,2,4,5,6 NOD2 CpG DNA, viral ssRNA, NOD-like R dsRNA NOD1
Endosome MyD88 Virus dsRNA - TRIF Phagosome NFB IRFs RIG-I IFN MAPKs Cytoplasm RIG-like R TLR3,7/8,9 MDA 5 Toll-like Receptors (TLR) and their exogenous ligands TLRs recognize endogenous ligands (danger signals) Domain and oligomeric structure of mannose-binding lectin (MBL) and ficolins
Fujita, Nature Rev. Immunol. 2002 Green et al Immunity 2007: Mammalian N-glycan branching protects against innate immune self-recognition and inflammation in autoimmune disease pathogenesis. Constitutive and inducible humoral sensors: cellular sources
Microbes-damaged tissues
Cellular sensors: TLR and coreceptors; NLR; RLR; capturing R
PMN DC Mø Endothelium Epithelia min hr days
PTX3 PTX3, C, Pentraxins (CRP, SAP, PTX3) Properdin M-Ficolin, Collectins (C1q, SP-A, SP-D, MBL) Ficolins SAA3 Ficolins, SAAs
C activation; opsonization; agglutination and neutralization; regulation of inflammation The complement cascade Classical pathway Lectin pathway Alternative pathway
Immune complexes, Carbohydrates C3b-microorganisms Pentraxins
C1q MBL, ficolins
C1r, C3b C1s MASPs Factor B, Factor D, C4, C2 Properdin C3 and C5 convertases C3a, C5a C3b Factor H
C5b-C9 Amplification loop
C1q
Inflammation Membrane Opsonization Ficolin attack complex, lysis MBL Receptors for humoral PRM and interplay with cellular innate immunity
CRP/SAP PTX3 C1q MBL SP-D SP-A Ficolins
CRT CRT
gC1q-R/p33 CR1 CR1 C1qRp CD91/CRT C1qRp CD91/CRT SIRP CD91/CRT (CD35) (CD35) FcR (CD93) (CD93)
+ - TLR SIGNALING; NFkB
OPSONIZATION THE PENTRAXIN SUPERFAMILY
(Garlanda, Bottazzi and Mantovani Annu Rev Immunol 2005) THE PROTOTYPIC LONG PENTRAXIN PTX3
Human Genomic clone 17 Kb Chr 3q25 Murine gene Chr 3
mRNA 1866 bp
Protein 381 aa Protomer 43 kDa Signal X domain Pentraxin domain Glycosylation 2 kDa peptide
Protein is mainly organized in covalently linked octamer
Inforzato et al, JBC 2008 (Garlanda et al Annu Rev Immunol 2005; Bottazzi et al Curr Op Immunol, 2006, 2008) CELLULAR SOURCES and FUNCTIONS OF PTX3
GDF9 TLR AGONISTS PRIMARY INFLAMMATORY OXIDIZED IL-10 FSH, cAMP, EGF (LPS. OmpA, CpG..) CYTOKINES (IL-1, TNF) LDL (+ oocyte)
PMN Mø myeloid Fibroblasts adipocytes mesangial Endothelium Granulosa DC Decidual cells Epithelium cells stromal cells SMC
MATRIX FERTILITY DEPOSITION Cumulus oophorus and maturation ANGIOGENESIS Fertilization PTX3 Decidualization and Interaction with placentation TSG-6 and II. Implantation Interaction with FGF2 INNATE IMMUNITY Resistance to A. Modulation of fumigatus, INFLAMMATION and P. brasiliensis, COMPLEMENT ACTIVATION P. aeruginosa, Interaction with C1q, Factor H, (Garlanda Bottazzi and Mantovani K. pneumoniae, Annu Rev Immunol 2005; Bottazzi Ficolin-2, apoptotic cells, P-selectin. CMV, Influenza virus... Curr Op Immunol 2006) Cooperation among soluble and cell-associated PRR COMPLEXITY AND COMPLEMENTARITY OF CELLULAR AND HUMORAL INNATE IMMUNITY
Microbe SCAVENGER R DECTIN-1 CD14 PMN TLR min
Microbe
hrs CellularCellular MØ/DC innate innate PTX3 immunityimmunity
HumoralHumoral amplificationamplification of of innateinnate immunity immunity Cytokines, C activation, chemokines, opsonization, other effector molecules agglutination (Jeannin, Bottazzi et al Immunity, 2005; Jaillon et al J Exp Med 2007) INNATEINNATE IMMUNE IMMUNE RESPONSE RESPONSE PENTRAXINS (CRP, SAP, PTX3)
Classical pathway Lectin pathway Alternative pathway Immune complexes, Carbohydrates C3b-microorganisms Pentraxins C1q MBL, ficolins
C1r, C3b C1s MASPs Factor B, Factor D, C4, C2 Properdin C3 and C5 convertases C3a, C5a C3b Factor H
C5b-C9 Amplification loop C1q Ficolin
MBL
CRP Inflammation Membrane Opsonization attack complex, SAP lysis PTX3 Role of Complement and FcReceptors in the protective activity of the long pentraxin PTX3 against Aspergillus fumigatus Complement Alternative pathway
Relevance in vivo CR3-CD11b FcR-CD32
Moalli et al. 2010 Model for PTX3-dependent opsonisation of A. fumigatus conidia
In the presence of PTX3-opsonized conidia, FcRIIa induces inside-out CD11b/CD18 activation, recruitment to the phagocytic cup and amplification of C3b-opsonized conidia phagocytosis. Moalli et al. 2010 Microbes-damaged tissues
Cellular sensors: TLR and coreceptors; NLR; RLR; capturing R
DC Mø PMN Endothelium
min hr days
epithelia
Activated Mø, PMN, DC
PTX3 PTX3, C, Pentraxins (CRP, SAP, PTX3) Properdin M-Ficolin, Collectins (C1q, SP-A, SP-D, MBL) Ficolins SAA3 Ficolins, SAAs Cytokines, chemokines, other effector molecules Constitutive and inducible humoral sensors
C activation; opsonization; agglutination and Synergism neutralization; regulation of inflammation - regulation Cellular innate immunity Humoral, fluid phase, innate immunity The inflammatory response
PTX3
CRP The IL-1 receptor (ILR) and Toll-like receptor (TLR) superfamily
ILRs TLRs
- IL-1F5 IL-1F5 - IL-1ra IL-1F6 IL-1 IL-1F8 - IL-1F7 LPS, CpG, ??? IL-1 IL-1 IL-33 IL-1F9 IL-18 IL-18 Flagellin, Poly I:C... I I 2 I P R T 1 R P B R R R 2 R - 1 R 0 P P S P L R 1 R / - 8 1 1 1 / c 1 - p P 1 c 8 R c c - P 8 - - - 8 R L 1 r 1 - L A 1 c I I - A 1 R L A L L A I L A R 1 T - I I I I I R - I A L G L I R 1 R I L G T I - I I 1 S L - L T I T L I - - -
MyD88 TIRAP/MAL Ig domain Leucine - rich repeat TRIF TIR domain domain SARM TRAM - Negative regulation TIR8/SIGIRR: ? Unknown ligand The decoy paradigm: Wald et al, Nature Immunol, 2003 Colotta et al, Science, 1993 Garlanda et al, PNAS, 2004 Mantovani et al, Trends Immunol 2002 Garlanda et al. Trends Immunol. 2009 Production and Release of IL-1β
From Host defence and Tissue homeostasis To Destructive inflammation: Auto-inflammatory diseases, Gout, RA and Juvenile arthritis, Osteoarthritis, Ischemia-reperfusion injury, Type 2 diabetes, Multiple myeloma, Multiple Sclerosis… IL-1 itself induces IL-1secretion in sterile inflammation and auto-inflammatory diseases.
Dinarello, Cell 2010 Inflammasome-independent processing of pro-IL-1β
In addition to caspase-1-dependent activation, pro-IL-1βcan also be processed by neutrophil-derived serine proteases, or pathogen-derived proteases. Netea MG, PLoS Pathogen 2010 Cellular sources of interleukin-1 family members and their effects on innate immune cells.
Cytokine release can be elicited in response to external stimuli (for example, Toll-like receptor activation in macrophages) or passively (owing to tissue damage or other forms of cell necrosis). Only IL-1 is known to be produced by B and T cells. Each of the IL-1 family cytokines acts on an overlapping range of innate immune cells
Sims JE, Nat. Rev. Immunol. 2010 The effects of interleukin-1 family members on CD4+ T cells.
IL-1 promotes the proliferation and survival of naive T cells and is crucial for the development of the TH17 cell subset. By cooperating with STAT3- inducing cytokines, IL-1 induces RORγt expression and IL-1R expression to reinforce TH17 cell function. IL-18 and IL-33 cooperate with STAT4- and STAT5- inducing cytokines to reinforce the TH1 cell and TH2 cell subsets, respectively. IL-1 family members drive effector cytokine production in the absence of T cell receptor signals, thus providing a mechanism for antigen- independent T cell immune responses. Sims JE, Nat. Rev. Immunol. 2010 TIR8 protein organization
? IL-1F5
Human (AF113795): •a single extracellular Ig domain Chromosome 11 band p15.5 (aa 17-112) 10 exons (11700 bp) mRNA: 1695 bp •5 potential N-glycosylation sites Protein: 410 aa
Mouse: •a transmembrane domain Chromosome 7 band F4 (aa 117-139) 9 exons (4000 bp) C222- •an intracellular TIR domain (aa 166-305) mRNA: 1450 bp TIR Protein: 409 aa L305- •2 conserved aa (Ser447 and Tyr536), replaced by Cys 222, Leu 305 TIR8 and tir8: 82% aa identity •95 aa long intracellular tail Increased acute and chronic intestinal inflammation and colon carcinogenesis in the absence of Tir8
(Garlanda et al. Cancer Res. 2007) (Garlanda et al. Cancer Res. 2007)
Regulation of colitis-associated cancer by TIR8/SIGIRR
IL-1ra LPS
IL-1 Epithelial cells, iDC,
Endothelial cells T T T T T T T T AcP I I I IL-1RI I TIR8/ TLR4 I I I I TIR8/ R R R R SIGIRR R R R R SIGIRR
D D MyD88 D MyD88 D TIR8+/+ - - TIR8-/-
Pro-inflammatory cytokines Inflammatory chemokines
IL-10 TGF-
IFN
Macrophage infiltration TIR8/SIGIRR prevents murine lupus by suppressing the immunostimulatory effect of lupus autoantigens
massive lymphoproliferation Immunopathology
) B cell proliferation s
b Peribronchial 1.5 A inflammation ( lpr +/+
B6 /Tir8
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r 0.5 p
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B Y12 Y12-U1snRNP
Immunocomplex DC activation deposition 20
Y12-U1snRNP
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m 10 Complement / g
n deposition ( 5
0
4 0
p 20
2 U1snRNP + DOTAP 1
- 15
L lpr lpr -/-
I B6 B6 SIGIRR 10
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0 B6lpr B6lpr B6lpr Tir8+/+ Tir8+/- Tir8-/-
Lech et al. JEM 2009 Regulatory function of TIR8/SIGIRR in pathology
Human: Chromosome 11, 10 exons mRNA: 1695 bp TIR domain Protein: 410 aa Mouse: IL-1 LPS Chromosome 7, 9 exons IL-18 CpG mRNA: 1450 bp IL-1F5 IL-33 nucleosomes Death domain Protein: 409 aa ??? ??
Conserved aa (S447, Y536)
Replaced aa TIR8/ IL-1RI TLR4 (C222, L305) SIGIRR IL-18R TLR9 T1/ST2 TLR7 Multiple sclerosis ?? MyD88 Kidney transplantation IRAK Pulmonary TB
Autoimmunity Fungal infections NFkB Asthma Arthritis Intestinal inflammation Colitis – associated cancer
Garlanda et al. Trends Immunol. 2009 xrclua oanmyitreewt eeoieiaino IL of heterodimerization with trapping interfere by may possibly domain TIR8, extracellular of portion intracellular the requires IL IL (e.g. antagonists receptor Unlike - 8P,TR sangtv euao hc fet h,T2adT and Th2 Th1, affects which regulator negative a is TIR8 18BP), euaino elplrzto yIL by polarization cell T of Regulation -
nlmaoydiseases inflammatory chronic and Autoimmune IL-1RII - h7T2Th Th2 Th17
IL-1RI IL IL IL - - - 1beta 1ra
IL-1R 1 AcP alpha -
TIR8/ SIGIRR - r rIL or 1ra Allergy -
F)addcyrcposo idn rtis(IL proteins binding or receptors decoy and 1F7) T1/ST2
IL-1R AcP IL - 33 - TIR8/ SIGIRR IRAK - R n IL and 1R1 - IL-18BP - 1 ifrnito.Inhibition differentiation. h17 - n TRAF and 1 IL-18R - Rfml members family 1R IL IL 1 - - 18 1
IL-18R F7 alnae l rnsImnl 2009 Immunol. Trends al. et Garlanda - 1AcP.
- AcP
TIR8/ -
,wiethe while 6, SIGIRR - RIor 1RII Istituto Clinico Humanitas/ University of Milan University of Perugia Barbara Bottazzi Luigina Romani Giuseppe Peri Antonio Inforzato University of Palermo Livija Deban Francesco Dieli Andrea Doni Yeny Martinez INSERM, Anger, France Federica Moalli Sebastien Jaillon Nadia Polentarutti Pascale Jeannin Cecilia Garlanda Manchester University Anthony Day Alberto Mantovani
Istituto Mario Negri Leiden University Antonio Bastone Mohamed Daha Roberto Latini Monica Salio University of Helsinki, Helsinki, Finland Seppo Meri University of Turin Emilio Hirsch University of Copenhagen, Copenhagen, Denmark Munchen University Peter Garred J-H Anders Functional genomics approaches in animal models to study human immunological disease
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