Filogenesi dei PRR. Sviluppo dorsoventrale della drosophila e per resistenza ai funghi
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1 Filogenesi dei PRR (NF-κB) Sviluppo dorsoventrale della drosophila e per resistenza ai funghi TIR: Toll/IL-1 receptor domain (dominio di interazione proteina-proteina)
2 TLR-like receptor signalling
3 The two arms of the Immune Response: APC
4 Homing and trafficking of DC during their maturation CCR7-/CCR5+ CCR7+/CCR5- mature DC Molecules expressed by matured DC PPRs
5 Switch CK-R durante maturazione DC Maturazione delle cellule dendritiche PPRs NF-κB
6 Vaccine adjuvants target the immune system to enhance the immunogenicity of coadminister antigens
7 Toll-like receptors recognize a range of pathogenderived products HMG1 O'Neill LA. Nat. Immunol. 2005
8 Apetoh L. et al. Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Nat Med TLR4-/- mice non sono sensibili a effetto tumorigenico di chemioterapici. SNP di TLR4 su 280 breast cancer patients (mutazione asp Glu), porta a perdita di funzione e mancata risposta a chemioterapia Danger signals endogeni: HMGB1, DNA mitocondriale, HSP, IL-1β.
9 NKT DC B NK CD8+ DC CD4+ DLN φ CD4+ CD4+ CD4+ CD4+ CD4+ CD8+ CD8+ CD8+ CD8+ CD8+ Immunosurveillance B IL10 VEGF MMP9 TGFβ PGE 2 IDO IL13 MICA/B PI-9 NK MDSC CD8+ CD4+ MDSC Treg CD8+ DLN MDSC NKT Treg Treg DC DC Treg CD8+ CD4+ MDSC DC CD8+ NKT CD4+ CD8+ φ CD8+ CD8+ NK CD8+ NK CD8+ CD4+ CD4+ NKT IFNγ IFNα TRAIL Fas NK CD4+ CD8+ MDSC = NK cell = CD4+ T cell = CD8+ T cell φ = MAC DC NKT = DC = NKT cell MDSC Treg B = MDSC = Treg cell = plasma cell
10 (beta-galactoside) From Dunn et al. Immunity 21, 2004, Pages
11 Immunotherapy Unbalance towards immune response
12 Active and passive Immunotherapy
13 -high MHC class II -costimulatory molecules -high CCR7
14 Cancer immunotherapy via dendritic cells. Palucka K, Banchereau J. Nat Rev Cancer. 2012
15 Vaccination Coley s Mixed Toxins (Streptococcus pyoegenes/pinibanil/ok-432 (TLR2/3), Serratia marcescens, TLR) Bacillus Calmette-Guerin (bladder cancer). Oggi: CpG (TLR9 ligand)
16 Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
17 Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
18 Antigen (1): Whole tumor cell or extracts Advantages: No need to know the exact antigen Easy to apply when using tumor cell lines Disadvantages Possible induction of autoimmunity
19 Antigen (2): Defined tumor antigens Tumor antigens: Tumor-specific: expressed only in tumor cells Tumor-associated: overexpressed in tumor cells Advantages Reduction of possible autoimmune induction Disadvantages Need to know the exact sequence of the antigen
20
21 Studi clinici nell'uomo di vaccinazione anti-tumorale con cellule dendritiche TUMORE TIPO di ANTIGENE TIPO di DC Melanoma gp100, MART1,MAGE-3 peptidi tirosinasi Differenziate da CD34 + MART1,MAGE-1 e 3 peptidi gp100 Differenziate da monociti lisati tumorali Mieloma multiplo Ig idiotipiche Differenziate da CD34 + Ca prostatico peptidi PSMA Differenziate da Monociti Ca renale, mammella lisati tumorali Ca polmone CEA Differenziate da monociti Ca colon e pancreas
22 Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
23 Adjuvant Activate the immune system. Different adjuvants allow the activation of different types of immune response. Prolong the time of release of the antigen
24 Adjuvants target Dendritic cells Vaccine: Antigen+Adjuvant Immature DC Draining Lymph node LMPH (6-12 hours) Mature DC
25 DC induce a broad immune cell activation CD8+ T cells Cytotoxic T cells IL-2 Perforin/ granzyme MHC I IFN-g NO IL-12 mature DC MHC II?? CD4+ T cells Th1 Fas IFN-g Th2 IL-4 B cells NK cells
26 Adjuvants (1): act at different levels on DCs Antigen + Adjuvant TLRs 1) Activation Bacterial products TLRs, Activating receptors antigen processing pathway DC DC activation 2) Antigen processing MHC I vs MHC II MHC II MHC I Immune synapsis CD8 CD4 TCR T cell CD40 B7-1/2 CD40L CD28 cytokines 3) T cell polarization IL-12 Th1 IL-4 Th2 signal 1 antigen specificity signal 2 costimulation signal 3 Th1/Th2 polarization
27 Adjuvants (2): determine the outcome of IR Mature Dendritic cell High costimulation High adhesion Immature Dendritic cell No costimulation Naive T PRIMING EXPANSION POLARIZATION ANERG / TOLERANCE Effector T EFFECTOR FUNCTION EXPANSION EFFECTOR FUNCTION DEATH
28 Adjuvants (2): influence ag processing and T cell activation Antigen + Adjuvant Antigen + Particulate Adjuvant (mineral salts, oil/water emulsions, microparticles: ) NALP3-IL-1β Activation and recruiment of various leukocyte populations DC Targeting C-type lectin R on DC (DNGR-1) allows cross-presentation -ISCOMATRIX (cholesterol, phospholipids and saponin from Quillaa Saponaria) MHC II CD4 TCR MHC I CD8 Th CTL Th 2 Th 1 Humoral response Cellular response
29 Inflammasome and autoinflammatory syndromes Hoffman HM et al. Lancet 364: , 2004
30 Ennio De Gregorio et al Current Opinion in Immunology 2009
31 TLR4-/- mice non sono sensibili a effetto tumorigenico di chemioterapici. SNP di TLR4 su 280 breast cancer patients (mutazione asp Glu), porta a loss of function e mancata risposta a chemioterapia Chemoterapia meno efficace in NALP3-/- mice e in trattamenti con anti-il-1β IL-1β è importante per funzioni di CD8+ T cells, visto in sarcomi sperimentali HMG1 è trigger per IL-1β
32
33 Problemi
34 Studi clinici nell'uomo di vaccinazione anti-tumorale con cellule dendritiche TUMORE TIPO di ANTIGENE TIPO di DC Melanoma gp100, MART1,MAGE-3 peptidi tirosinasi Differenziate da CD34 + MART1,MAGE-1 e 3 peptidi gp100 Differenziate da monociti lisati tumorali Mieloma multiplo Ig idiotipiche Differenziate da CD34 + Ca prostatico peptidi PSMA Differenziate da Monociti Ca renale, mammella lisati tumorali Ca polmone CEA Differenziate da monociti Ca colon e pancreas
35 Apetoh L. et al Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy Nat. Med. 2007
36
37 Classi di Pattern Recognition Receptors ante-antebodies: Dominio molecolare tipo collagenico. Riconoscono strutture circolanti di funghi e batteri Riconoscono cellule apoptotiche. Controllano autoimmunità RIG-I-like receptor (RLR) es: TLR9: sequenze CpG, tpiche i del DNA ipometilato dei batteri IFN-dependent- TRIF-IRF3
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