Infl ammasomes: Intracellular Regulators of Pathogen Recognition, Host Defense, and Infl ammation Mechanisms to Reduce IL-1β or IL-18 Signaling P IL-1 & IL-18 Signaling Pathways Infl ammasome-mediated -1 Activation Regulates the Secretion of IL-1β & IL-18 IκB Inhibitors Mechanisms of Action Nod-like receptors (NLRs) are a subset of pattern recognition receptors (PRRs) found in the that are essential for detecting invading pathogens and initiating the innate immune response. IL-18 BP IL-18 binding (IL-18 BP); Binds to IL-18 with higher a nity than either the bound or soluble NLRs are activated either by bacterial, fungal, or viral molecules that contain pathogen-associated molecular patterns (PAMPs), or by nonmicrobial danger signals (DAMPs) released from damaged IKK forms of IL-18 R and prevents IL-18 signaling IL-1ra An IL-1 receptor antagonist that prevents IL-1 from binding to its receptor cells. Upon activation, some NLRs oligomerize to form multiprotein in ammasome complexes that serve as platforms for the recruitment, cleavage, and activation of in ammatory . P IL-1 RII A decoy IL-1 receptor with a short cytoplasmic domain that is incapable of transducing an IL-1 signal In ammasome oligomerization requires two signals, a priming signal that results in the transcription of Pro-IL-1β and Pro-IL-18 (–-—), and a second signal that promotes indirect IκB IL-1β Signaling SIGIRR Single immunoglobulin IL-1-related receptor (SIGIRR); Prevents IL-1 RI/IL-1 RAcP heterodimerization activation of the in ammasome, such as ion or membrane perturbations, reactive oxygen species (ROS), or ATP (˜). In ammasome oligomerization leads to the activation of Caspase-1, followed NFκB . activation Soluble IL-1 RI or RII Soluble receptors that can bind to IL-1 and IL-1 RAcP, but are incapable of propagating a signal TAB1/2 by the maturation and secretion of IL-1β and IL-18 (™-›), and in some cases, an in ammatory form of cell death known as . In ammasome/Caspase-1-dependent secretion of IL-1NADβ FIND CARD . Recruitment of in ammatory cells A soluble receptor that can bind to IL-1/IL-1 RI, but is incapable of propagating a signal; Enhances IL-1 NAD FIND CARD TAK1 TRAF-6 . Induction of secondary cytokines, enzymes, Soluble IL-1 RAcP binding to soluble IL-1 RII and IL-18 stimulates the in ammatory response by inducing the expression of secondary mediators that promote the recruitment of immune cells to the site of the infection (œ). In addition, IL-18 in ammatory mediators, and acute phase that regulate the local and systemic response enhances the cytolytic activity ofx naturalAD killerNACHT (NK) cells and promotes IFN-γ production. To date, four in ammasome complexes (NLRP1, NLRP3, IPAF, and AIM2) have been partially characterized.NADx ADFIND NACHTCARD IL-18 Rβ NAD FIND CARD IL-1 RI These complexes contain a speci c NLR family protein or AIM2, the ASC and/or Cardinal adaptor proteins, and Pro-Caspase-1. JNK IL-18  IL-18 Signaling x PYD BIR AD NACHT x PYDAD BIRNACHT TRAF-6 Although the secretion of IL-1β and IL-18 are intended to combat infection, constitutive in ammasome activation and the subsequent overproduction of IL-1β or IL-18 can have detrimental e ects P IL-1β  IL-18 Rα P MyD88 IL-1 RAcP that are associated with autoin ammatory and autoimmune disorders. For these reasons, mechanisms that inhibit IL-1β and IL-18 signaling are of interest. Decoy or soluble receptors that sequesterNAD FIND CARD MKK IL-18 BP PYD BIRNAD FIND CARD κ IRAK1/2 IRAK4 IL-1β, non-signaling IL-1β antagonists, and disruption of IL-1 receptor heterodimerization are intrinsic pathways that inhibit IL-1β signaling. Similarly, naturally occurring IL-18 bindingPYD proteinBIR NF B AP-1 IL-1 RAcP NAD  x NAD FIND CARD x FIND CARD (IL-18 BP) can prevent IL-18 from bindingAD toNACHT its receptor. Further research is necessary to characterize how in ammasome complexes are activated, the mechanisms by which IL-1β and IL-18 AD NACHT p38 signaling can be regulated, and both the bene cial and detrimental e ects associated with the in ammasome pathway. These  ndings may have therapeutic implications for in ammasome- œ IL-1 RI IL-1 Signaling x AD NACHT x AD NACHT IL-1ra related disorders, including autoin PYD ammatoryBIR disorders, Crohn’s disease, vitiligo, gout, asbestosis, and Alzheimer’s disease.  PYD BIR Inflammatory PYD BIR Response IL-1β IL-1 RAcP NADPYD BIR DOMAIN ORGANIZATIONNAD FIND CARD OF THE NLR PROTEINS OLIGOMERIZED COMPLEXES FIND CARD IL-18 Signaling Active IL-1β IL-1 RII  Human Name Mouse Name Family CARD-containing NLRs NLRP1 (NALP1) Infl ammasome Select Microbial Activators IL-18 Rα IL-1β x AD NACHT Pro-Caspase-1 Pro-Caspase-5 x NADAD FINDNACHTCARD . Induction of IFN-γ in T cells and NK cells Decoy   CIITA (NLRA) CIIta (Nlra) NAD FIND CARDNLRA NLRP1/ASC/Pro-Caspase-1/Pro-Caspase-5 NAD FIND CARD . Induction of secondary pro-in ammatory NAD FIND CARD NAD FIND CARD cytokines, chemokines, cell adhesion molecules IL-18 SIGIRR NAD FIND CARD NAD FIND CARD x NAD BIR FIND CARD NAD FIND CARD . Induction of , NO synthesis IL-18 Rβ NOD1 (NLRC1) Nod1 (Nlrc1)PYDNADAD FINDNACHTCARDNLRC Pro-Caspase-5 Pro-Caspase-1 Pro-Caspase-1 Pro-Caspase-5 x ADBIR NACHT x AD NACHT ASC NLRP1  lethal toxin PYDx AD NACHT Active IL-18 IL-1 RI x NADAD FINDNACHTCARD  Muramyl dipeptide (MDP) x NAD FIND CARD NOD2 (NLRC2) Nod2 (Nlrc2)x AD NACHT NLRC x AD NACHT Soluble ADBIR NACHT x AD NACHT PYD ASC NLRP1 IPAF (NLRC4) Ipaf (Nlrc4)PYDx BIR NLRC Pro-Caspase-5 Pro-Caspase-1 ASC NLRP1 PYD BIR IL-1 RI or RII PYD ADBIR NACHT Pro-Caspase-1 PYDx ADBIR NACHT PYD BIR PYD BIR Human Name Mouse NamePYD BIR Family BIR-containing NLRs Pro-Caspase-1 Pro-Caspase-5 PYD BIR IL-1 RAcP Pro-Caspase-1 NLRP3 (NALP3) Infl ammasome Select Microbial Activators PYD BIR Signal 2 PYD BIR ASC NLRP1 K+ DAMP/PAMP IL-1 RI NAIP Naip1-7 NLRB Pro-Caspase-1NLRP3/ASC/Cardinal/Pro-Caspase-1Pro-Caspase-1 Pro-Caspase-5 PYD BIR Inflammasome Activation K+ Soluble Cardinal ASC NLRP3 ˜ Bacterial Type III or Human Name Mouse Name Family PYD-containingPro-Caspase-5 NLRsPro-Caspase-1 ASC NLRP1  Adenovirus, Bacterial RNA ATP IL-1 RAcP CardinalPro-Caspase-1ASC NLRP3  Candida albicans/Saccharomyces cerevisiae IV Secretion Systems + Pore-forming NLRP1 (NALP1) Nlrp1a-c (Nalp1) NLRP Pro-Caspase-5 Pro-Caspase-1 ASC NLRP1  Danger signals: ATP, NAD , β-amyloid › Cardinal ASC NLRP3 Signal 1 Toxins Secretion of Nlrp2, Nlrp3 (Nalp3), Nlrp4a-g ASC NLRP1 and particulates such as calcium pyrophosphate Pannexin-1 Particulate Pro-Caspase-1Pro-Caspase-1 P2X7 Receptor β NLRP2-9 (NALP2-9), (Nalp4a-g), Nlrp5, Nlrp6, Cardinal ASC NLRP3 dihydrate and monosodium urate Activators IL-1 & IL-18 NLRP Pro-Caspase-1ASC NLRP1 – NLRP11-14 (NALP11-14) Nlrp9a-c (Nalp9a-c), Nlrp12 Pro-Caspase-1Pro-Caspase-1 Pro-Caspase-5  Xenogenous compounds: Silica, asbestos, and alum (Nalp12), Nlrp14 (Nalp14) Pro-Caspase-1  In uenza Endogenous Cytokine Active IL-1β Pro-Caspase-1 Active IL-18 IPAF IPAF  Listeria monocytogenes PAMP NLRP10 (NALP10) Nlrp10 (Nalp10) NLRP Pro-Caspase-1Pro-Caspase-1 CardinalPro-Caspase-1ASC NLRP3Pro-Caspase-5 Microbial Ligand Pro-Caspase-5 Pro-Caspase-1 Pro-Caspase-1ASC NLRP1Pro-Caspase-5  Lipopolysaccharide (LPS) Human Name Mouse Name FamilyCardinalUnknownASC N-terminalNLRP3 Domain Pro-Caspase-1 Pro-Caspase-5  Muramyl dipeptide NAD(MDP) FIND CARD ROS NAD FIND CARD Pro-Caspase-1 IPAF CardinalPro-Caspase-1Pro-Caspase-1ASC NLRPPro-Caspase-53 NAD FIND CARD NLRC3NAD (NOD3), FINDNLRC5CARD IPAF Pro-Caspase-1IPAF IPAF Pro-Caspase-5  Sendai virus IL-1 RAcP Phagosome Nlrc3, Nlrc5 NLRC Cardinal ASC NLRP3 (NOD27) ASC Pro-Caspase-5ASC NLRP1Pro-Caspase-1 ASC NLRP1  Staphylococcus aureus x AD NACHT Pro-Caspase-5 Pro-Caspase-1 Pro-Caspase-1Pro-Caspase-1ASC Pro-Caspase-1NLRPPro-Caspase-51 x x Pro-Caspase-1Pro-Caspase-5Pro-Caspase-1NAIP Pro-Caspase-1 x AD NACHT ROS NLRX1 (NOD9)AD NACHT Nlrx1 NLRX Pro-Caspase-5IPAF Pro-Caspase-1 Pro-Caspase-1ASC NAIPNLRP1 AD NACHT IL-1 RI TIR Domain Pro-Caspase-5 Pro-Caspase-1 ASC IPAF NLRP1 Pro-Caspase-1 IPAF (NLRC4) InflASC ammasomePro-Caspase-1NLRP1 Select Microbial Activators Death Domain BIRDOMAIN ORGANIZATION OF RELATEDASC ADAPTORASC PROTEINSPro-Caspase-1NLRP1 PYD Pro-Caspase-5ASC NLRP1Pro-Caspase-1 Pro-Caspase-1 Phagolysosome PYD BIR Pro-Caspase-1ASC IPAFNLRP1 IPAF/ASC/Pro-Caspase-1Pro-Caspase-1ASC IPAFNLRP1 PYD BIR ASC: -associated speck-like protein containing a CARD ASC NAIP IPAFNLRP1 Cardinal ASC NAIP NLRP3 PYD BIR Toll-like Receptor MyD88 ASC NLRP1 Pro-Caspase-1 TIRAP Pro-Caspase-1 IPAF Pro-Caspase-1Pro-Caspase-1 IPAF Cardinal Pro-Caspase-1Pro-Caspase-1ASC NLRPIPAF3 IPAF MyD88 Cardinal: CARD inhibitor of NFκB-activating ligands (found only in humans) Pro-Caspase-1ASC NLRP1 Pro-Caspase-1 IPAF  Pseudomonas aeruginosa TRAM Membrane Perturbations Pro-Caspase-1Pro-Caspase-1 Pro-Caspase-1 ASCPro-Caspase-1IPAF Cardinal ASC NLRP3  Salmonella typhimurium TRIF ™ β Pro-Caspase-1 IPAF Cardinal ASCPro-Caspase-1NLRP3 Mature IL-1 2+ Cardinal ASC CardinalPro-Caspase-1ASCPro-Caspase-1NLRP3IPAF  Shigella exneri Pro-Caspase-1ASC NLRPNAIP3 Cardinal ASC IPAF NLRP3 Inflammasome Ca DOMAIN KEY Cardinal ASCASCPro-Caspase-1NLRP3 Cardinal ASC NAIPNLRP3 NLRP3 In ammasome Cardinal ASC NLRP3 Oligomerization Cardinal Pro-Caspase-1ASC NLRP3 Protein-Protein Interacting Domains Cardinal ASC NLRPNAIP3 ASC Pyroptosis Mature IL-18 ASC Cardinal ASC Pro-Caspase-1NLRPNAIP3 MKK Caspase Recruitment Domain (CARD) Pro-Caspase-1 IPAF IPAF Cardinal ASC Pro-Caspase-1NLRPIPAF3 Pro-Caspase-1 ASCPro-Caspase-1AIM2 IPAF/NAIP/Pro-Caspase-1Pro-Caspase-1 š (PYD) Pro-Caspase-1Pro-Caspase-1 Pro-Caspase-1Pro-Caspase-1Pro-Caspase-1 Pro-Caspase-1Pro-Caspase-1 ASC Pro-Caspase-1IPAF Pro-Caspase-1Pro-Caspase-1IPAF IPAF Pro-Caspase-1 Cytokine Processing 2+ Baculovirus Repeat Domain (BIR) IPAF Pro-Caspase-1 IPAF IKK Ca Pro-Caspase-1Pro-Caspase-1IPAFIPAF ASC IPAFIPAF IPAF  Legionella pneumophila JNK Unknown N-terminal Domain ASCAIM2 ASCPro-Caspase-1IPAF Pro-Caspase-1ASCAIM2 IPAFIPAF IPAF Pro-Caspase-1 Pro-Caspase-5 IPAF -Binding/Oligomerization Domain Pro-Caspase-1 NAIPIPAFIPAF Pro-Caspase-1 Pro-Caspase-5 IPAF Pro-IL-18 Pro-Caspase-1 Pro-Caspase-1 NAIPIPAF Pro-Caspase-1ASC IPAFIPAF Pro-Caspase-1 NAIP, CIITA, HET-E, TP-1 Domain (NACHT)ASCPro-Caspase-1 IPAFIPAF Pro-Caspase-1ASC IPAFIPAFIPAF Pro-Caspase-1 IPAF ASCPro-Caspase-1AIM2 IPAFIPAF p38 Pro-IL-1β Pro-Caspase-5 Pro-Caspase-1 ASCPro-Caspase-1 IPAF Pro-Caspase-1 IPAF Active Caspase-1 Microbial Ligand Recognition Domain ASC NAIP ASC NLRPAIM21 Infl ammasome IPAF Select Microbial Activators Proteasome Pro-Caspase-5 Pro-Caspase-1 ASCPro-Caspase-1Pro-Caspase-1IPAF ASC NLRP1 ASC NAIP Leucine-Rich Repeat Domain (LRR) ASCPro-Caspase-1ASCAIM2 NAIP Pro-Caspase-1 P NAIPIPAF Pro-Caspase-1AIM2/ASC/Pro-Caspase-1NAIPIPAF NAIP Pro-Caspase-1 NAIP Hematopoietic -inducible Nuclear Protein with aNAIP 200 a.a. repeat (HIN200) NAIPIPAF ASC NLRP1 ASC ASCAIM2 Pro-Caspase-1NAIP P Other Domains ASC NLRP1 Pro-Caspase-1 IPAF Pro-Caspase-1 IκB Pro-Caspase-1 NAIP Pro-Caspase-1 IκB Pro-Caspase-1 IPAFIPAF ASCAIM2 NAIP  DS-DNA from virus, , or the host itself AP-1 NACHT-Associated Domain (NAD) Pro-Caspase-1ASC IPAFIPAF Pro-Caspase-1 IPAF Pro-Caspase-1 Pro-Caspase-1 IPAF Pro-Caspase-1 IPAFIPAF NFκB FunctionPro-Caspase-1 to Find Domain (FIIND) IPAF Pro-Caspase-1ASCAIM2 NFκB Pro-Caspase-1 Pro-Caspase-1 IPAFIPAF Transcription of Activation Domain (AD) Pro-Caspase-1 Pro-Caspase-1ASC IPAF ASC CardinalIPAF ASC NLRP3 IPAF IL-1β and IL-18 ASC Cardinal ASC NLRPOligomerized3 Pro-Caspase-1in ammasome complexes may consist of more subunits than what is shown. Cardinal ASC NLRP3 ASC IPAF Cardinal ASC NLRP3 Pro-Caspase-1ASCASCAIM2 Pro-Caspase-1 — This illustration represents general processes suggested in the scienti c literature and is Pro-Caspase-1notASC to be considered comprehensive nor de nitive. ©2010,ASC 2015 R&D Systems Pro-in ammatory Cytokines Pro-Caspase-1 ASC MP_In ammasomes_1550 Pro-Caspase-1ASC ASC Pro-Caspase-1 Pro-Caspase-1 ASC Pro-Caspase-1 ASCAIM2 Pro-Caspase-1 Pro-Caspase-1ASC Pro-Caspase-1 ASCAIM2 Pro-Caspase-1ASCAIM2 Global [email protected] [email protected] Pro-Caspase-1 ASCAIM2 Pro-Caspase-1ASC North America TELASC 800AIM2 343 7475 Pro-Caspase-1 IPAF IPAF Europe | Middle East | Africa TEL +44 (0)1235IPAF 529449 ASCAIM2 LEARN MORE IPAF ASCAIM2 Pro-Caspase-1 China ASCAIM2 rndsystems.com/ [email protected] TEL +86 (21) 52380373 ASCAIM2 Pro-Caspase-1 IPAF ASCAIM2 inflammasome Pro-Caspase-1 Rest of World bio-techne.com/fiPro-Caspase-1 nd-us/distributorsIPAF TEL +1 612 379ASC 2956AIM2 IPAF Pro-Caspase-1 IPAF bio-techne.com ASC ASC ASCAIM2 NAIP NAIP NAIP NAIP

Pro-Caspase-1 IPAF Pro-Caspase-1 IPAF Pro-Caspase-1 IPAF Pro-Caspase-1 IPAF

ASC ASC Pro-Caspase-1 ASC Pro-Caspase-1 ASC ASCAIM2 Pro-Caspase-1 ASCAIM2 Pro-Caspase-1

ASCAIM2 ASCAIM2