Dendritic cells. Emmanuel STEPHEN-VICTOR (PhD) INSERM U Centre de Recherche des Cordeliers, Paris
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1 Dendritic cells Emmanuel STEPHEN-VICTOR (PhD) INSERM U Centre de Recherche des Cordeliers, Paris
2 Pathogens Bacteria, virus, fungi The pathogen-derived antigens need to be transported adptaive immune response needs to be initiated
3 Immunity and Anti-immunity Host Vs Pathogen Bacteria, virus, fungi Mф Pathogens Th1 Th2 Neutrophils DC Priming adaptive immune response CD4 T Th17 Tregs NET Innate immune cells Adaptive immunity
4 Dendritic cells -Discovery 1868 Paul Langerhans => Branched skin cells at epidemis resembling neurons
5 Dendritic cells -Discovery 1868 Paul Langerhans => Branched skin cells at epidemis resembling neurons 1973 Ralph Steinman => Dendritic cells from the spleen of mice
6 One of the first views of DCs in mouse spleen Steinman RM et al. J Exp Med, 1973
7 Dendritic cells: Major professional antigen presenting cells Critical for primary immune response Large array of endocytic receptors (not B cells) Large array of antigen presenting and co-stimulatory molecules (not MØ) Ability to migrate to T cell areas for presentation (not other APCs)
8 Some features of dendritic cells Adhesion and co-stimulation ICAM 1,3; B7-1, B7-2; CD58/LFA3 Secretion IL-12, IL-1, TNF, chemokines High levels of Ag presenting molecules HLA I, II; CD1 Receptors for Ag uptake MMR, DC25, Langerin, integrins, FcγR TLR, CR Signaling of DC TNF-R, CD4, TLR Cytokine R: GM-CSF, IL-4, IL1, TGF-b, IL-1 Receptors for pathogens CD4, DC-SIGN, chemokine receptors
9 Dendritic cells in health and disease Immunological Reviews Volume 219, Issue 1, pages , 1 SEP 27 DOI: /j.16-65X x
10 Ontogeny of dendritic cells
11 Origin and development of dendritic cells LC pro LC MP: Myeloid progenitor, MDP: monocyte dendritic cell progenitor, CDP: committed DC progenitor Flt3L: fms-like tyrosine kinase 3 ligand Liu & Nussenzweig, Immunol Rev 21
12 Subsets of dendritic cells
13 Dendritic cell subsets in health and disease Immunological Reviews Volume 219, Issue 1, pages , 1 SEP 27 DOI: /j.16-65X x
14 Human dendritic cell deficiency Collin et al., Nature Rev Immunol 211
15 Purification / preparation of dendritic cells DCs Low population <1% Spleenocytes <1% blood leukocytes <1% tissues (skin, lungs) Obtaining DC 1992 J Banchereau et C Caux (Lyon) Bone marrow cells DC human 1992 R Steinman (NY) K Inaba (Kyoto) Splenic cells DC murine 1994 A Lanzavecchia (Bellinzona, Suisse) Blood monocytes DC or macrophages
16 Host immune response to pathogens Dendritic cell CD4+ Th cell B cell Antibodies Humoral/antibody response Pathogens Bacteria, fungi
17 MHC Class II presentation by Dendritic cells invariant chain (Ii) MHC class II compartment (MIIC) class II-associated Ii peptide (CLIP) Neefjes et al 211
18 Host immune response to pathogens Dendritic cell CD4+ Th cell B cell Antibodies Humoral/antibody response Pathogens Intracellular bacteria, virus CD8+ Tc cell CTL response
19 MHC Class I presentation by Dendritic cells TAP:transporter associated with antigen presentation, ER: endoplasmic reticulum Neefjes et al 211
20 CD4+T cell immune response to pathogens Antigen Antigen PRR PRR PRR APC
21 A. fumigatus AIR Airborne fungal spores Up to 1 9 /m 3 Innate immune cell Healthy Immunocompromised Elimination by phagocytosis Invasive aspergilosis SOIL
22 A. fumigatus sensing by innate cells Role of Rodlet Rodlet layer Innate cell Rodlet layer degraded Dormant conidia Enlarged or Germinated conidia Immune response Inert Immune response Activated Aimanianda*, Bayry* et al Nature. 29
23 A. fumigatus structural organization of the conidia Electron micrograph of fixed conidia PAMPS Chitin Courtesy: Beauvais A Rodlet layer Polysaccharides Proteins Lipids Exposure of polysaccharides during germination
24 PRRs on dendritic cells
25 PRRS and DCs 211 Noble prize Bruce Beutler Jules Hoffmann Ralph Seinman Chitin Exposure of polysaccharides during Toll receptors germination Dendritic cells
26 Changes during dendritic cell maturation Immature DC Mature DC
27 Changes during dendritic cell maturation Immature DC Pathogens Pro-inflammatory cytokines T cells Mature DC High intracellular HLA II High endocytosis, Low CD54, 58, 8, 86 Low CD4, Low CD83 Low granule antigens High surface HLA II Low endocytosis High CD54, 58, 8, 86 High CD4, High CD83 Production of inflammatory cytokines
28 Methodology PBMC CD14 Monocytes GM-CSF & IL-4 5 days MoDC A. fumigatus (morphotypes) Dormant, Enlarged, Germinated conidia 48 hrs DC responses
29 Ctr DOC EC GC Ctr DOC EC GC CD8 MFI CD86 MFI IL-6 (pg/ml) IL-12 p7 (pg/ml) HLA-DR MFI CD4 MFI IL-8 (pg/ml) IL-1b (pg/ml) Enlarged and germinated conidia but not dormant conidia induce maturation and activation of human dendritic cells 32 * * * * * * * * * * * * * * * *
30 Migration of dendritic cells to draining lymph nodes
31 CD4+T cell immune response to pathogens Cytokines SIGNAL 3 Cytokine receptor Transcription Factors Cytokines APC CD54 (ICAM-1) CD4 MHC II + Ag Peptide SIGNAL 2 SIGNAL 1 CD11a/CD18 (LFA-1) CD4L CD4 T cell TCR CD3 complex Th1 Th2 T-bet STAT 4 STAT 1 GATA3 STAT 6 IFN- IL-2 IL-4 IL-5 IL-13 Effector cells CD8/86 CD4 CD28/CTLA-4 Th17 RORC2 STAT 3 IL-17 IL-17F IL-21 IL-22 CD58 (LFA-3) SIGNAL 2 CD2 itreg FoxP3 STAT 5 TGF-b IL-1 suppresors Cytokine receptor Cytokines SIGNAL 3
32 Dendritic cells in health and disease Immunological Reviews Volume 219, Issue 1, pages , 1 SEP 27 DOI: /j.16-65X x
33 Regulation of Dendritic cell functions by immune compartment
34 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins Cytokines Regulatory T cells
35 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins Cytokines Regulatory T cells
36 None RO RA None RO RA T cells induce activation of dendritic cells Co-stimulatory molecules CD8 CD MFI
37 None RO RA None RO RA None RO RA pg/ml pg/ml ng/ml T cells induce activation of dendritic cells Inflammatory cytokines TNF-a IL-6 IL
38 Stimulation of dendritic cells by CD4L (CD154) on T cells leads to maturation HLA-DR CD11c CD86 CD8 CD4 Anti-CD154/CD4L Isotype Ctrl
39 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins Cytokines Regulatory T cells
40 Dendritic cells and regulatory T cells CD4+CD25 + regulatory T cells in human circulation Represent ~2-4% of total PBMCs Express CD25 FoxP3 (transcription factor forkhead) Intracellular CTLA-4 Expression Immunosuppressors
41 CD4+CD25 + regulatory T cells: Mechanisms of suppression Vignali et al., 28; Nat Rev Immunol
42 CD4+CD25 + regulatory T cells: Mechanisms of suppression Vignali et al., 28; Nat Rev Immunol
43 None Treg RO RA None Treg RO RA CD4+CD25+ regulatory T cells inhibit TLR-mediated activation of human dendritic cells Co-stimulatory molecules 8 * CD8 * * 12 * CD86 * * 6 9 MFI J Immunol 24 J Immunol 27 Arthritis Rheum 27 Nature Rev Immunol 28 Nature 28 Arthritis Rheum 29 Nature Rev Rheumatol 29 Am J Pathol 29 Vaccine 211
44 None Treg RO RA None Treg RO RA None Treg RO RA pg/ml pg/ml ng/ml CD4+CD25+ regulatory T cells inhibit TLR-mediated activation of human dendritic cells Inflammatory cytokines TNF-a IL-6 IL * * 9 6 * * 4 3 * J Immunol 24 J Immunol 27 Arthritis Rheum 27 Nature Rev Immunol 28 Nature 28 Arthritis Rheum 29 Nature Rev Rheumatol 29 Am J Pathol 29 Vaccine 211
45 MFI MFI MFI MFI MFI MFI CTLA-4 is the major molecule implicated in the CD4+CD25+ regulatory T cell-mediated inhibition of dendritic cells CD83 CD CTLA-4 (1µg) - + CTLA-4 (1µg) - + CD HLA-DR CTLA-4 (1µg) - + CTLA-4 (1µg) - + CD4 CD1a CTLA-4 (1µg) - + CTLA-4 (1µg) - +
46 CD4+CD25+ regulatory T cells inhibit T cell proliferation mediated by TLR-activated human dendritic cells Poly IC DC-Treg Poly IC DC-RO CFSE Bayry et al., J Immunol. 27
47 pg/ml ng/ml pg/ml CD4+CD25+ regulatory T cells inhibit DC-mediated IFNresponses IFN- TNF-a IL-2 Poly IC DC Treg RO IL-5 IL-4 IL-1 Bayry et al., J Immunol. 27
48 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins Cytokines Regulatory T cells
49 Regulation of Dendritic cell functions by immune compartment A IL-8 * IL-6 * Type I interferon: Promotion of activation of DC TNF-a IL-1b * GM-CSF + IL-4 GM-CSF + IL-4 + IFNb1a B CCL19 CCL22 CCL17 Maddur et al., Arthrits Rheum 21
50 MFI MFI MFI MFI Regulation of Dendritic cell functions by immune compartment 75 CD83 6 CD TGF-β: inhibit the DC maturation TGF-β (1ng) - + TGF-β (1ng) CD86 6 HLA-DR TGF-β (1ng)- + TGF-β (1ng)- +
51 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins Cytokines Regulatory T cells
52 Regulatory B cells slow down the complete maturation of idcs into mdcs. B cell stimulation: CD4L+CpG-ODN by American Society of Hematology HLA-DR CD8 CD86 Morva A et al. Blood 212;119: Maddur M S et al. Blood 212;119:
53 Activated B cells induce dendritic cell maturation and induce Th2 polarization by inducing OX-4 ligand Activation Resting B cells Activated B cells Immature DCs Maturation of DCs OX4L Mature DCs Maddur et al., Nature Commun 214
54 Activated B cells induce dendritic cell maturation and induce Th2 polarization by inducing OX-4 ligand Activation Resting B cells Activated B cells Immature DCs B cell help Maturation of DCs IL-4 IL-13 IL-5 GATA3 Th2 cells Differentiation OX4L Naïve CD4 + T cells Mature DCs Maddur et al., Nature Commun 214
55 Regulation of Dendritic cell functions by immune compartment Effector CD4+ T cells DC B cells / Immunoglobulins IgG immunoglobulins at therapeutic high doses (IVIg) Cytokines Regulatory T cells
56 Regulation of dendritic cells by circulating antibodies: Intravenous Immunoglobulin (IVIg) Intravenous Immunoglobulin (IVIg) Therapeutic preparation of pooled normal polyspecific human IgG obtained from large numbers of healthy donors Contains Antibodies to non-self (foreign antigens) Antibodies to self antigens Antibodies to antibodies (anti-ids)
57 Intravenous Immunoglobulin (IVIg) Polyclonal IgG plasma pool thousands of healthy donors Primary Immunodeficiencies CVID, XLA Secondary Immunodeficiencies BM transplantation, Multiple myeloma Autoimmune and inflammatory diseases Immune thrombocytopenic purpura (ITP) Guillain-Barré syndrome (GBS) Kawasaki disease (KD) CIDP Dermatomyositis (DM) Polymyositis (PM) Psoriasis Systemic lupus erythematosus (SLE) ANCA-associated vasculiits Myasthenia gravis (MG) Transplant rejection Wide - divergent therapeutic applications
58 pg/ml Regulation of maturation and function of dendritic cells by IVIg at therapeutic concentration IVIg 6 (8) 15 (21) CD1a 43 (39) 1 (27) CD IL-12 * IVIg 4 IL-1 * Ctl IVIg HSA 2 Immature Mature Blood 23, Arthritis Rheum 23, PNAS 29
59 Contribution of DC to autoimmunity Genetic Epigenetic Environmental Cytokines/chemokines Ag presentation Co-stimulatory molecules Migratory properties Effector /pathogenic T cells Treg number and function
60 Dendritic cells prevent autoimmunity by expansion of regulatory T cells
61 Regulatory T cell expansion by dendritic cells Transforming growth factor-β (TGF-β) Cyclooxygenase-2 (COX-2) Indoleamine 2,3-dioxygenase (IDO) Treg Inducible costimulatory ligand (ICOS-L) CD252 (OX4L) Programmed death ligands Pallotta et al., Nat Immunol. 211 Trinath et al., Blood 213 Gopisetty et al., Immunology 213 Fantini et al., Immunology 24 Sakaguchi et al., Cell, 28 Dejaco et al., Immunology, 27
62 % positive dendritic cells Regulatory T cell expansion by dendritic cells: Role of Programmed death ligands 1 * Ctr Mtb Ctr Mtb Ctr Mtb PD-L2 PD-L1 PD-1 Trinath et al. J Infect Dis 212
63 % CD4+CD25+FoxP3+ T cells Regulatory T cell expansion by dendritic cells: Role of Programmed death ligands 15 * * 1 5 Ctr Mtb Mtb+ anti-pd-l1 Trinath et al. J Infect Dis 212
64 Ctr IVIg 1 mg IVIg 15 mg HSA Ctr IVIg 1 mg IVIg 15 mg HSA PGE 2 (pg/ml) Regulatory T cell expansion by dendritic cells: Role of PGE2 5 4 * * 3 COX-2 OX-2 -actin β-actin 2 1 Ctr IVIg 1 mg IVIg 15 mg HSA Ctr IVIg 1 mg IVIg 15 mg HSA Trinath et al Blood 213
65 Ctr IVIg NS IVIg Ctr IVIg NS IVIg PGE 2 (pg/ml) % CD4 + FoxP3 + T cells Regulatory T cell expansion by dendritic cells: Role of PGE * * * * * 1 2 Ctr IVIg IVIg + COX-2 inhibition Ctr IVIg IVIg + COX-2 inhibition Trinath et al Blood 213
66 Differentiation of itregs by dendritic cells Comparison of different cell populations for the induction of CD4+CD25+ FOXP3+ Treg's. Banerjee D K et al. Blood 26;18:
67 Take home message Immature/tolerogenic DC Activated DC Immune tolerance Prevention of autoimmunity Induction of Tregs Maintenance/homeostasis of Tregs T and B cell response to pathogens Clearance of pathogens Anti-tumor immunity
68 Take home message Immature/tolerogenic DC Activated DC Immune tolerance Prevention of autoimmunity Induction of Tregs Maintenance/homeostasis of Tregs T and B cell response to pathogens Clearance of pathogens Anti-tumor immunity Genetic Epigenetic Environmental Autoimmunity Inflammatory diseases
69 Thank you
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