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)
TLR-like receptor signalling
The two arms of the Immune Response: APC
Homing and trafficking of DC during their maturation CCR7-/CCR5+ CCR7+/CCR5- mature DC Molecules expressed by matured DC PPRs
Switch CK-R durante maturazione DC Maturazione delle cellule dendritiche PPRs NF-κB
Vaccine adjuvants target the immune system to enhance the immunogenicity of coadminister antigens
Toll-like receptors recognize a range of pathogenderived products HMG1 O'Neill LA. Nat. Immunol. 2005
Apetoh L. et al. Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Nat Med. 2007 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.
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
(beta-galactoside) From Dunn et al. Immunity 21, 2004, Pages 137-148
Immunotherapy Unbalance towards immune response
Active and passive Immunotherapy
-high MHC class II -costimulatory molecules -high CCR7
Cancer immunotherapy via dendritic cells. Palucka K, Banchereau J. Nat Rev Cancer. 2012
Vaccination Coley s Mixed Toxins (Streptococcus pyoegenes/pinibanil/ok-432 (TLR2/3), Serratia marcescens, TLR) Bacillus Calmette-Guerin (bladder cancer). Oggi: CpG (TLR9 ligand)
Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
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
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
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
Parameters for efficient immunotherapy 1. Choice of antigen 2. Choice of adjuvant 3. Correct type of induced immune response 4. Induction of immunological memory
Adjuvant Activate the immune system. Different adjuvants allow the activation of different types of immune response. Prolong the time of release of the antigen
Adjuvants target Dendritic cells Vaccine: Antigen+Adjuvant Immature DC Draining Lymph node LMPH (6-12 hours) Mature DC
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
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
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
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
Inflammasome and autoinflammatory syndromes Hoffman HM et al. Lancet 364:1779-1785, 2004
Ennio De Gregorio et al Current Opinion in Immunology 2009
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
Problemi
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
Apetoh L. et al Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy Nat. Med. 2007
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