dendritic cells · 2019-12-24 · activation of human dendritic cells co-stimulatory molecules j...

Dendritic cells

Emmanuel STEPHEN-VICTOR (PhD)

INSERM U 1138 - Centre de Recherche des Cordeliers, Paris

Pathogens

Bacteria, virus, fungi

The pathogen-derived antigens need to be transported

adptaive immune response needs to be initiated

Innate immune cells

NET

Adaptive immunity

DC

Neutrophils

Mф

Priming adaptive

immune response

Immunity and Anti-immunity – Host Vs Pathogen

Bacteria, virus,

fungi

Pathogens

CD4 TTregs

Th1

Th17

Th2

Priming adaptive

immune response

Dendritic cells -Discovery

1868 Paul Langerhans => Branched skin cells at epidemis resembling neurons

Dendritic cells -Discovery

1868 Paul Langerhans

=> Branched skin cells at epidemis resembling neurons

1973 Ralph Steinman

=> Dendritic cells from the spleen of mice

One of the first views of DCs in mouse spleen

Steinman RM et al. J Exp Med, 1973

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)

Dendritic cells: Major professional antigen presenting cells

Some features of dendritic cells

Adhesion and co-stimulation

ICAM 1,3; B7-1, B7-2; CD58/LFA3

Receptors for pathogens

CD4, DC-SIGN, chemokine

receptors

Secretion

IL-12, IL-10, TNF, chemokines

High levels of Ag presenting

molecules

HLA I, II; CD1

Receptors for Ag uptake

MMR, DC205, Langerin, integrins,

FcγR TLR, CR

Signaling of DC

TNF-R, CD40, TLR

Cytokine R: GM-CSF, IL-4,

IL1, TGF-b, IL-10

Dendritic cells in health and disease

Immunological Reviews

Volume 219, Issue 1, pages 118-142, 10 SEP 2007 DOI: 10.1111/j.1600-065X.2007.00551.x

http://onlinelibrary.wiley.com/doi/10.1111/j.1600-065X.2007.00551.x/full#f1

http://onlinelibrary.wiley.com/doi/10.1111/imr.2007.219.issue-1/issuetoc


Ontogeny of dendritic cells

Origin and development of dendritic cells

MP: Myeloid progenitor, MDP: monocyte dendritic cell progenitor, CDP: committed DC progenitor

Liu & Nussenzweig, Immunol Rev 2010

Flt3L: fms-like tyrosine kinase 3 ligand

LC pro

LC

Subsets of dendritic cells

Dendritic cell subsets in health and disease






Collin et al., Nature Rev Immunol 2011

Human dendritic cell deficiency

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)

1992 R Steinman (NY)

K Inaba (Kyoto)

1994 A Lanzavecchia (Bellinzona, Suisse)

Bone marrow cells DC human

Splenic cells DC murine

Blood monocytes DC or macrophages

Pathogens

Dendritic cell CD4+ Th cell

B cell Antibodies

Humoral/antibody response

Bacteria, fungi

Host immune response to pathogens

invariant chain (Ii)

MHC class II compartment (MIIC)

class II-associated Ii peptide (CLIP) Neefjes et al 2011

MHC Class II presentation by Dendritic cells

Pathogens

Dendritic cell

CD8+ Tc cell

B cell Antibodies

Humoral/antibody response

CTL response

Intracellular bacteria , virus

CD4+ Th cell

Host immune response to pathogens

TAP:transporter associated with antigen presentation,

ER: endoplasmic reticulumNeefjes et al 2011

MHC Class I presentation by Dendritic cells

PRR

APC

PRR

PRR

Antigen

Antigen

CD4+T cell immune response to pathogens

AIR

SOIL

Up to 109/m3

Airborne fungal

spores

Innate

immune cell

Elimination by

phagocytosisInvasive

aspergilosis

Healthy Immunocompromised

A. fumigatus

Rodlet

layer

Immune response –

Inert

Immune response –

Activated

Enlarged or

Germinated conidia

Innate cell

Rodlet layer

degraded

Dormant conidia

A. fumigatus sensing by innate cells – Role of Rodlet

Aimanianda*, Bayry* et al Nature. 2009

A. fumigatus – structural organization of the conidia

Polysaccharides

Proteins

Lipids

Rodlet layer

Electron micrograph of fixed conidia

Courtesy: Beauvais A

Chitin

Exposure of polysaccharides during

germination

PAMPS

PRRs on dendritic cells

Chitin

Exposure of polysaccharides during

germination

PRRS and DCs – 2011 Noble prize

Bruce

BeutlerJules

Hoffmann

Ralph Seinman

Toll receptors Dendritic cells

Immature DC

Mature DC

Changes during dendritic cell maturation

Immature DC

High intracellular HLA II

High endocytosis,

Low CD54, 58, 80, 86

Low CD40,

Low CD83

Low granule antigens

High surface HLA II

Low endocytosis

High CD54, 58, 80, 86

High CD40,

High CD83

Production of inflammatory

cytokines

Mature DC

Pathogens

Pro-inflammatory cytokines

T cells

Changes during dendritic cell maturation

Methodology

CD14

Monocytes

MoDC

PBMC

GM-CSF & IL-4

5 days

DC responses

48 hrs

A. fumigatus

(morphotypes)

Dormant, Enlarged,

Germinated conidia

Enlarged and germinated conidia but not dormant conidia

induce maturation and activation of human dendritic cells

6000

0

4000

2000

* *

Ctr

DO

C

EC

GC

1800

0

1200

600

* *

CD

80

MF

I

Ctr

DO

C

EC

GC

1600

0

2400

800

3200 * *

HL

A-D

R M

FI

4000

0

6000

2000

8000

* *

CD

40 M

FI

CD

86

MF

I

IL-8

(p

g/m

l)

0

**

1000

15000

30000

45000

60000

IL-1b

(pg

/ml)

0

*

*

10150

300

450

600

IL-6

(p

g/m

l)

0

*

*

303000

6000

9000

12000 60

0

40

20

IL-1

2 p

70

(p

g/m

l)

*

*

Migration of dendritic cells to draining lymph nodes

CD54 (ICAM-1) CD11a/CD18 (LFA-1)

CD40 CD40L

MHC II + Ag Peptide TCR CD3 complex

CD80/86 CD28/CTLA-4

CD4

CD58 (LFA-3) CD2

APC CD4 T cellSIGNAL 1

SIGNAL 2

SIGNAL 2

SIGNAL 3

SIGNAL 3Cytokines

Cytokines

Cytokine receptor

Cytokine receptor

Th1

Th2

Th17

iTreg

T-bet

STAT 4

STAT 1

GATA3

STAT 6

RORC2

STAT 3

FoxP3

STAT 5

IFN-IL-2

IL-4

IL-5

IL-13

IL-17

IL-17F

IL-21

IL-22

TGF-b

IL-10

Transcription

Factors Cytokines

Effector

cells

suppresors

CD4+T cell immune response to pathogens

Dendritic cells in health and disease






Regulation of Dendritic cell functions by

immune compartment

DC

Effector CD4+ T cells

B cells / Immunoglobulins

Cytokines

Regulatory T cells


immune compartment

MFI

CD80 CD86

800

600

400

200

0

1200

900

600

300

0

RO

RA

No

ne

RO

RA

No

ne

Co-stimulatory molecules

T cells induce activation of dendritic cells

pg

/ml

0

30

20

0

TNF-a IL-8IL-6

900

600

300

10

ng

/ml

RO

RA

RO

RA

No

ne

No

ne

RO

RA

No

ne

pg

/ml

0

900

600

300

Inflammatory cytokines

T cells induce activation of dendritic cells

HLA-DR CD86 CD80 CD40CD11c

Stimulation of dendritic cells by CD40L (CD154) on T

cells leads to maturation

Isotype CtrlAnti-CD154/CD40L

DC



Cytokines


immune compartment

Regulatory T cells

•Represent ~2-4% of total PBMCs

•Express CD25

•FoxP3 (transcription factor forkhead)

•Intracellular CTLA-4 Expression

•Immunosuppressors

CD4+CD25+ regulatory T cells in human circulation

Dendritic cells and regulatory T cells

CD4+CD25+ regulatory T cells: Mechanisms of suppression

Vignali et al., 2008; Nat Rev Immunol

**

MFI

**

**800

600

400

200

0

1200

900

600

300

0

Tre

g

RO

RA

No

ne

Tre

g

RO

RA

No

ne

CD80 CD86

CD4+CD25+ regulatory T cells inhibit TLR-mediated

activation of human dendritic cells


J Immunol 2004

J Immunol 2007

Arthritis Rheum 2007

Nature Rev Immunol 2008

Nature 2008


Nature Rev Rheumatol 2009

Am J Pathol 2009

Vaccine 2011

pg

/ml

0

40

30

20

0

TNF-a IL-8IL-6

*

**

900

600

300

10

**

ng

/ml

Tre

g

RO

RA

Tre

g

RO

RA

No

ne

No

ne

Tre

g

RO

RA

No

ne

pg

/ml

0

900

600

300

Inflammatory cytokines

J Immunol 2004

J Immunol 2007


Nature Rev Immunol 2008

Nature 2008


Nature Rev Rheumatol 2009

Am J Pathol 2009

Vaccine 2011

CD4+CD25+ regulatory T cells inhibit TLR-mediated

activation of human dendritic cells

CTLA-4 is the major molecule implicated in the CD4+CD25+

regulatory T cell-mediated inhibition of dendritic cells

CTLA-4 (1µg) - + CTLA-4 (1µg) - +

CTLA-4 (1µg) - + CTLA-4 (1µg) - +

CTLA-4 (1µg) - + CTLA-4 (1µg) - +

CD83

0

200

400

600

MF

I

CD80

0

600

1200

1800

2400

MF

I

CD86

0

400

800

1200

1600

MF

I

HLA-DR

0

7000

14000

21000

28000

MF

I

CD40

0

7000

14000

21000

28000

MF

I

CD1a

0

6000

12000

18000

24000

MF

I

CFSE

Poly IC DC-Treg

100 101 102 103 104

Poly IC DC-RO

100 101 102 103 104

Bayry et al., J Immunol. 2007

CD4+CD25+ regulatory T cells inhibit T cell

proliferation mediated by TLR-activated human

dendritic cells

IFN- IL-2

ng

/ml

RO

Treg

Poly IC DC

0

65

130

0

150

300

TNF-ap

g/m

l

0

125

250

0

600

1200

IL-5

0

100

200

IL-4

0

40

80

IL-10

pg

/ml

Bayry et al., J Immunol. 2007

CD4+CD25+ regulatory T cells inhibit DC-mediated IFN-

responses

DC



Cytokines


immune compartment

Regulatory T cells

Regulation of Dendritic cell functions by immune

compartment

** IL-8 IL-6 A

CCL22 CCL17 CCL19

GM-CSF + IL-4 GM-CSF + IL-4 + IFNb1a

B

* *

*

TNF-a IL-1b

*

CCL3 CCL4

Type I interferon:

Promotion of activation of DC

Maddur et al., Arthrits Rheum 2010

Regulation of Dendritic cell functions by immune

compartment

TGF-β: inhibit the

DC maturation

TGF-β (10ng)- + TGF-β (10ng)- +

TGF-β (10ng) - + TGF-β (10ng) - +

TGF-β (10ng) - + TGF-β (10ng) - +

CD83

0

250

500

750

MF

I

CD80

0

2000

4000

6000

MF

I

HLA-DR

0

2000

4000

6000

MF

I

CD86

0

800

1600

2400

3200

MF

I

CD40

0

10000

20000

30000

40000

MF

I

CD1a

0

5000

10000

15000

20000

25000

MF

I

DC



Cytokines


immune compartment

Regulatory T cells

©2012 by American Society of Hematology

Morva A et al. Blood 2012;119:106-114

Maddur M S et al. Blood 2012;119:3863-3864

B cell stimulation:

CD40L+CpG-ODN 2006

Regulatory B cells slow down the complete maturation of iDCs

into mDCs.

HLA-DR CD80 CD86

Activated B cells induce dendritic cell maturation and induce

Th2 polarization by inducing OX-40 ligand

Resting B cells

Activation

Maturation of DCs

OX40L

Immature DCs

Mature DCs

Activated

B cells

Maddur et al., Nature Commun 2014

Activated B cells induce dendritic cell maturation and induce Th2 polarization by

inducing OX-40 ligand

Resting B cells

Activation

Maturation of DCs

Differentiation

OX40L

Th2 cells

B cell help

GATA3IL-4

IL-13 IL-5

Naïve

CD4+ T cells

Immature DCs

Mature DCs

Activated

B cells

Maddur et al., Nature Commun 2014


immune compartment

IgG immunoglobulins at

therapeutic high doses (IVIg)

DC



Cytokines

Regulatory T cells

• 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)

Regulation of dendritic cells by circulating

antibodies: Intravenous Immunoglobulin (IVIg)

Intravenous Immunoglobulin (IVIg)

Polyclonal IgG – plasma pool – thousands of healthy donors

Intravenous Immunoglobulin (IVIg)

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

Primary Immunodeficiencies

CVID,

XLA

Secondary Immunodeficiencies

BM transplantation,

Multiple myeloma

Wide - divergent therapeutic

applications

Regulation of maturation and function of dendritic

cells by IVIg at therapeutic concentration

60(80)

15(21) CD1a

43

(39)1

(27)CD83

IVIg

IVIg

Immature Mature

70

140

*

0

pg/m

L

*40

0

20

Ctl

IVIgHSA

IL-12

IL-10

Blood 2003, Arthritis Rheum 2003, PNAS 2009

Contribution of DC to autoimmunity

Genetic

Epigenetic

Environmental

Cytokines/chemokines

Ag presentation


Migratory properties

Effector /pathogenic T cells

Treg number and function

Dendritic cells prevent autoimmunity by

expansion of regulatory T cells

Treg

Transforming growth factor-β (TGF-β)

Cyclooxygenase-2 (COX-2)

Indoleamine 2,3-dioxygenase (IDO)

Inducible costimulatory ligand (ICOS-L)

CD252 (OX40L)

Programmed death ligands

Regulatory T cell expansion by dendritic cells

Pallotta et al., Nat Immunol. 2011

Trinath et al., Blood 2013

Gopisetty et al., Immunology 2013

Fantini et al., Immunology 2004

Sakaguchi et al., Cell, 2008

Dejaco et al., Immunology, 2007

Trinath et al. J Infect Dis 2012

Regulatory T cell expansion by dendritic cells: Role of Programmed death ligands

0

25

50

75

100

PD-L2 PD-L1 PD-1

Ctr Mtb Ctr Mtb Ctr Mtb

*

% p

osit

ive d

en

dri

tic c

ells

Trinath et al. J Infect Dis 2012

Regulatory T cell expansion by dendritic cells: Role of Programmed death ligands

0

5

10

15

Ctr Mtb Mtb+anti-PD-L1

* *

% C

D4+

CD

25+

Fo

xP

3+

T c

ells

Regulatory T cell expansion by dendritic cells: Role of PGE2

Trinath et al Blood 2013

Ctr

IVIg

10

mg

IVIg

15

mg

HS

A

COX-2

! -actin

Ctr

IVIg

10 m

g

IVIg

15 m

g

HS

A

0

1000

2000

3000

4000

5000* *

PG

E2 (

pg

/ml)

Ctr IVIg

10 mg

IVIg

15 mg

HSA Ctr IVIg

10 mg

IVIg

15 mg

HSA

COX-2

β-actin

Regulatory T cell expansion by dendritic cells: Role of PGE2

Ctr

IVIg

NS

-39

8 +

IV

Ig

0

2

4

6

8**

% C

D4

+F

oxP

3+ T

cells

Ctr IVIg IVIg +

COX-2 inhibition

Ctr

IVIg

NS

-39

8 +

IV

Ig

0

1000

2000

3000

4000

*

**

PG

E2 (

pg

/ml)

Ctr IVIg IVIg +

COX-2 inhibition

Trinath et al Blood 2013

Comparison of different cell populations for the induction of CD4+CD25+ FOXP3+ Treg's.

Banerjee D K et al. Blood 2006;108:2655-2661

Differentiation of iTregs by dendritic cells

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

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

Take home message

Immature/tolerogenic DC Activated DC

Autoimmunity

Inflammatory diseases

Genetic

Epigenetic

Environmental

Thank you

dendritic cells · 2019-12-24 · activation of human dendritic cells co-stimulatory molecules j...

Documents