1. CO-STIMULATION IS ESSENTIAL FOR PRIMING OF NAIVE T LYMPHOCYTES The antigen-specific and the co-stimulatory signal has to be induced in concert to induce T lymphocyte activation The antigen-specific and co-stimulatory signals can be delivered simultaneously by professional antigen presenting cells, only The antigen-specific and the co-stimulatory singnals has to be delivered by the same professional antigen presenting cell

2. PROFESSIONAL ANTIGEN PRESENTING CELLS Express MHC class I and class II molecules Express co-stimulatory molecules (B7, CD40) Take up extracellular antigens B cells – soluble proteins, toxins (ADAPTIVE) Macrophages – extracellular pathogens (bacteria, yeast) INNATE – particles Dendritic cells – viruses, apoptotic cells

3. A DENDRITIKUS SEJTEK BIOLÓGIÁJA Fertőző betegségek Szervátültetés Tumorok eliminálása Gatekeeper funkció Kórokozók „érzékelése” Adaptív immunválasz elindítása Saját struktúrákkal szembeni tolerancia fenntartása

4. CHARACTERISTICS OF PROFESSIONAL ANTIGEN PRESENTING CELLS Macrophage Dendritic cellB - lymphocyte Ag uptake phagocytosis +++ phagocytosis +++ Ag-specific mIg virus infection ++++ ++++ MHC expression induced +/+++ constitutive ++++ constitutive +++ bacteria, cytokine immature/mature +++/++++ activation ++++ Pesented Ag particulate Ag protein soluble protein intra/extracellular virus protein, allergen toxin pathogens apoptotic cell Co-stimulation induced +/++ constitutive ++++ induced +/+++ éretlen/érett +++/++++ Localization lymphoid tissue lymphoid tissue lymphoid tissue connective tissue connective tissue peripheral blood body cavities epithelium Lymph node evenly immature – tissue follicles mature – T cell area

5. PROFESSIONAL ANTIGEN PRESENTING CELLS Express MHC class I and class II molecules Express co-stimulatory molecules (CD40, B7) Take up extracellular antigens B cells – soluble proteins, toxins ADAPTIVE – Ag specific Macrophages – extracellular pathogens (bacteria, yeast) Dendritic cells – viruses, apoptotic cells INNATE 3 – 6% ~1% ~25%

6. MYELOID LYMPHOID BONE MARROW CMPCLP DC2/plasmacytoid IPC/preDC2 Pre-Tα Vλ5 CD34+ CLA+ CD11c+ CD1a+ Langerhans/LC CD34+ CLA- CD11c+ CD1a- Interstitial BLOOD TISSUE Macrophage CD14+ Monocyte/preDC1 DC1/myeloid CD34+ HPC ORIGIN AND DIFFERENTIATION OF HUMAN DENDRITIC CELLS

7. TRIF TANK IKKεTBK1 IRF-3 TRIF TRAM TLR3 TLR4 MyD88 IRF-5 TLR7 TLR8 TLR9 IFN-β IFN-α1 RIG-1 Ig production by B cells is induced Type I interferon receptor IRF-7 Enhanced NK cell cytotoxic activity Activation of  and γδ T cells Cross-presentation by conventional dendritic cells is enhanced IRAK-1 TRAF-6 IRF-7 PLASMACYTOID DENDRITIC CELLS AS PROFESSIONAL TYPE I INTERFERON SECRETING CELLS Vírus infection

8. IDC TISSUE ACTIVATION Antigen uptake Activation Mobility Antigen processing & presentation MHCI/MHCII MDC  DC1/DC2 LYMPHOID ORGANS DC – T interaction Cognate Th/Tc activation Th1/Th2 instruction HUMAN MYELOID DENDRITIC CELLS

9. Morphology of plasmacytoid dendritic cells IPC/DC2 monocytepDC Scanning EMTransmission EM

10. Plasmacytoid DCs control the function of many immunocytes Role in immune response and in the pathogenesis of autoimmune diseases and cancer HIV infects PDC IFNα is impotant in SLE pathology

11. Tissue DC Activated DC DC AND T CELLS ENCOUNTER T CELL ACTIVATION CIRCULATION Naive T cells Effector and memory T cells TISSUE LYMPH NODE TISSUE Lymphatics Inflammation Pathogen ACTIVATION AND MIGRATION OF DENDRITIC CELLS ANTIGEN

12. Migration Pathways of PDC/IPC versus mDC into a lymph node IPC: HEV mDC: afferent lymphatics Both migrate into the T-cell rich areas

13. Macrophage Dendritic cell Activated macrophage Phagocytosis and degradation of backteria (LPS, TLR) DANGER SIGNAL Activated dendritic cell Virus, extracellular pathogens, inflammatory cytokines (LPS, TLR) DANGER SIGNAL Monocyte CHANGES OF TISSUE ENVIRONMENT INDUCES THE ACTIVATION OF MACROPHAGES AND DENDRITIC CELLS LYMPHOID TISSUEBLOODTISSUE

14. INTERDIGITATING RETICULAR (MATURE DENDRITIC) CELL IN T CELL AREAS OF LYMPH NODES NUCLEUS compact CYTOPLASM T CELL

15. Huang et al Immunity 2004 Bone-marrow derived DCs (yellow) were pulsed with 1 µM Ova 4 peptide and 10 µM Ova for 1 hour at 37 o C, then injected into the footpad of a C57BL/6 recipient. This was followed 6 hours later by i.v. co-injection of OT-I CD8+ T cells (5 µM CFSE, green) and OT- II CD4+ T cells (5 µM SNARF, red). Rapid DC Migration in the Subcapsular Space Capture of an Ag-Specific T Cell by an Ag-Bearing DC Bone-marrow derived DCs (either 5 µM CFSE, green) or (50 µM Cell Tracker Blue, blue) were injected into the footpad of a C57BL/6 mouse, followed 18 hours later by intravenous injection of freshly isolated polyclonal CD4+ T cells (5 µM SNARF, red) and CD8+ T cells (5 µM CFSE and 5 µM SNARF, yellow). The draining LN was removed 6 hours after injection

16. Activated dendritic cells act as professional antigen presenting cells MHC-peptide complexes1. signal STRANGER Co-stimulatory molecule 2. signal AMPLIFICATION Cytokines3. signal DANGER They are in close contact with specific T lymphocytes CONTACT OF DENDRITIC CELLS AND T - LYMPHOCYTES IN LYMPHOID ORGANS

17. ONE, TWO and DANGER signal hypothesis Matzinger, P. Science, 296. 301- (2002)

18. The danger hypothesis & co-stimulation Fuchs & Matzinger 1995 Full expression of T cell function and self tolerance depends upon when and where co-stimulatory molecules are expressed. Apoptotic cell death. A natural, often useful cell death. APC No danger Cell containing only self antigens Innocuous challenge to the immune system fails to activate APC and fails to activate the immune system

19. The danger hypothesis APC Necrotic cell death e.g. tissue damage, virus infection etc Pathogens recognised by microbial patterns DANGER APC that detect ‘danger’ signals express costimulatory molecules, activate T cells and the immune response

20. There is no window for tolerance induction in neonates Neonatal T cells are not intrinsically tolerisable but the neonatal environment predisposes to tolerance Antigens induce tolerance or immunity depending upon the ability of the immune system to sense them as ‘dangererous’, and not by sensing whether they are self or ‘non-self’. Apoptosis, the ‘non-dangerous’ death of self cells may prevent autoimmunity when old or surplus cells are disposed of. Suggests that tolerance is the default pathway of the immune system on encountering antigens. Explains why immunisations require adjuvants to stimulate cues of danger such as cytokines or costimulatory molecule expression. Some implications of the danger hypothesis Doesn’t exclude self-nonself discrimination, but is very hard to enequivocally disprove experimentally

21. LOCALIZATIONANTIGEN UPTAKE MHC LOCALIZATION ANTIGEN PRESENTATION ANTIGEN/ PATHOGEN B-cellLymphoid tissue, Peripheral blood BCR-mediated endocytosis, pinocytosis MCII/CIIV Cell surface (activation) low antigen dose co-stimulation toxins, víruses, bacteria, any protein MacrophageLymphoid tissue, connective tissue, body cavities phagocytosis Fc  R, CR MCII Cell surface (activation) high antigen dose co-stimulation intracellular bacteria, other pathogens particles Immature DC Epithelium, skin, tissues phagocytosis, macropinocytosi s, pinocytosis intracellular MCII nonevíruses, allergens, bacteria, lipids, any protein Maturing DC afferent lymphNot significantintracellular CIIV inefficient Mature DCLymph node T-cell areas noneCell surfacevery efficient co-stimulation PROFESSIONAL ANTIGEN PRESENTING CELLS