mechanisms of danger- signal mediated immune modulation
TRANSCRIPT
The Self-Non-Self theory
• Dominant model in immunology since the 1950s • The body is able to discern between self and non-self• Thus an immune response is triggered against all foreign entities• No immune response is triggered against an organism’s endogenous
entities
Danger Theory• Rooted in Janeway’s work (Infectious non-self
theory)• Proposed by Polly Matzinger in a 1994 article title
“Tolerance, Danger and the Extended Family”• States that the immune response is a result of the
organism reacting to the emission of “danger signals” by the organism, not “non-self” entities• Self constituents can trigger an immune response
if they are dangerous (cellular stress) and non-self constituents can be tolerated (commensal bacteria)
Predications made by Self-non-self, Infectious non-self theory and Danger Theory
Burnet 1969 Janeway 1989 Matzinger 1994
DAMPs – Damage signal criteria
• Should be active as a highly purified molecule• Biological activity should not be due to contamination with microbial
molecules (LPS)• Should be active at concentrations that are actually present in
pathophysiological situations• Selective elimination or inactivation of a DAMP should ideally inhibit
the biological activity of dead cells (in vitro and vivo)
Molecular identification of Danger Signals• Cellular stress – when a cell is stressed, even in the absence of any foreign
substance, it emits molecules that activate APCs• Heat-shock proteins – expression increased with elevated temperature and
other stresses, can bind antigen and activate APCs• Necrotic cell death – intracellular contents, including damage-associated
molecular patterns (DAMPs). Apoptosis?• Uric Acid – released by injured cells, dendritic cell maturation, with antigen it
enhances respsonses from CD8+ cells• High-mobility-group box 1 – signals damage, initiates inflammatory
response/repair • Inflammasomes?
Inflammasomes • Component of innate immunity, triggered by danger signals;
stress/infection• Multiprotein complex • Expressed in myeloid cells • Senses damage activate caspase1 production of IL-1β• Subsets – NLRP1, NLRP3 (codes Nalp3 inflammasome), NLRC4• Consists of caspase-1, caspase recruitment domain (CARD), NALP and
ASC (adaptor) • NALP – NOD like receptor that contains NACHT (nucleotide binding
domain), LRR and Pyrin domain
Malaria• Infects 300-500 million• Kills over 1 million children annually • Causative agent is a parasitic protozoan;
Plasmodium species • Complex life cycle involving a mosquito vector
and a human host • Erythrocyte (RBC) lysis resulting in fever, anemia
and death• 1-2% cases develop Cerebral Malaria (deadly)
Immune response and Plasmodium infection
Adaptive:• Induces an immune response characterized by IFNγ producing T cells• Production of antibodies against infected RBCs
Innate:• Several molecular conserved structures of Plasmodium act as pathogen-
associated molecular patterns (PAMPs)• PAMPs activate Toll-like receptors (TLRs) on macrophages and dendritic
cells• Hemozoin activation of a Nalp3 inflammasome
Hemozoin• Heme crystal formed by Plasmodium• During the intraerythrocytic cycle hemoglobin is digested• Results in free heme• Parasite is able to convert free heme into insoluble hemozoin crystals as a means of
detoxification• RBC lysis results in hemozoin entering the blood stream
Studies about the immuno-modulatory capacity are conflicting• Activation of TLR9 signaling• Dependence upon the presence of malarial DNA complexed to hemozoin• Inflammasome
Hypothesis
Hemozoin acts as a Nalp3 inflammasome activating danger signal resulting in IL-1β
production.
Hemozoin induces IL-1β secretion in myeloid cells
• Experiments used synthetic hemozin; β-hematin
• Bone marrow-derived macrophages (BMDM) produced low levels of TNF, IL-6 and MIP-1α with hemozoin, relative to CpG (TLR9 activator).
• BMDM robustly secreted IL-1β and IL-18 when primed and stimulated with HZ.
IL-1β HZ induction in THP1 cells and Murine BMDCs
THP1 cells: human macrophage like cell line
BMDCs: murine bone marrow-derived Dendritic cells
Hemozoin IL-1β secretion is independent from P2X7 activation
HZ induced IL-1B not mediated by ATP released from dying cells.
Uric acid crystals have no effect on hemozoin IL-1B, uric acid itself can act as danger signal
Toxic heme cannot activate caspase1, but is toxic (PARP cleavage)
Hemozoin IL-1β secretion is independent from MyD88-mediated
signaling pathways
Performed in order to remove any implication of DNA-mediated TLR9 signaling
Chloroquine – anti-malarial drug
Phagocytosis, K+ Efflux and activation of a NADPH oxidase are all essential for Hz-mediated
inflammasome
Phagocytosis, K+ Efflux and activation of a NADPH oxidase are all essential for Hz-mediated
inflammasome
Phagocytosis, K+ Efflux and activation of a NADPH oxidase are all essential for Hz-mediated
inflammasome
Conclusion• Were able to show that Malarial Hemozoin is a Nalp3 inflammasome
activating signal, therefore enhancing the pro-inflammatory activity along with TLRs• May lead to novel more efficient anti-malaria drugs• Investigate the mechanism for inflammasome activation by agonists
and therefore the exact role of hemozoin• Self-non-self vs Danger? Maybe a combination of both.