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Immune tolerance
Lecture outline
• Principles of immune regulation
• Self-tolerance; mechanisms of central
and peripheral tolerance
• Inhibitory receptors of T cells
• Treg’s and IL-2
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The immunological equilibrium: balancing
lymphocyte activation and control
Activation Tolerance
Effector T cells Regulatory T cells
Normal: reactions Controlled response to
against pathogens pathogens
Inflammatory No response to self
disease, e.g. reactions
against self
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Central and peripheral tolerance to self
The principal fate
of lymphocytes that
recognize self antigens
in the generative organs
is death (deletion), BUT:
Some B cells may change
their specificity (called
“receptor editing”)
Some T cells may
differentiate into
regulatory (suppressor)
T lymphocytes
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 7th edition, 2011 c Elsevier
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Consequences of self antigen recognition in thymus
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 7th edition, 2011 c Elsevier
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What self antigens are seen in the
thymus?
• Ubiquitous cell-associated and circulating
proteins
• The thymus has a special mechanism for
displaying peripheral tissue antigens in
thymic medullary epithelial cells, where
they signal self-reactive thymocytes for
death
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Consequences of AIRE mutation
• Human disease: autoimmune
polyendocrinopathy with candidiasis and
ectodermal dysplasia (APECED), also called
autoimmune polyendocrine syndrome (APS-1)
– Associated gene identified by positional cloning,
named AIRE (“autoimmune regulator”)
• Mouse knockout: autoantibodies against
multiple endocrine organs, retina
– Failure to express many self antigens in the thymus
--> failure of negative selection
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Deletion of self-reactive T cells in the thymus:
how are self antigens expressed in the thymus?
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 8th edition, 2014
AIRE (autoimmune regulator) is a regulator of gene transcription
that stimulates thymic expression of many self antigens which are
largely restricted to peripheral tissues
[Link] GW/+ mice develop
peripheral neuropathy (CIDP)
Sciatic Nerve
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Peripheral T cell tolerance
Abbas, Lichtman and Pillai. Basic Immunology, 4th edition, 2014
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T cell anergy
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 7th edition, 2011 c Elsevier
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CTLA-4 competitively inhibits B7-CD28
engagement
APC APC
B7 B7
CTLA-4
CD28
T Cell T cell (activated T
cell or Treg)
Costimulation à
T cell activation
CTLA-4 blocks and removes
B7 à lack of costimulation
à T cell inhibition
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The B7:CD28 families
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 8th edition, 2014
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Major functions of selected B7-CD28
family members
• CD28-B7: initiation of immune
Activation
responses
• ICOS-ICOS-L: T cell help in
germinal center reactions (antibody
responses)
• CTLA-4-B7: inhibits early T cell
Inhibition
responses in lymphoid organs
• PD-1:PD-L1,2: inhibits effector T
cell responses in peripheral tissues
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Blocking CTLA-4 promotes tumor rejection:
CTLA-4 limits immune responses to tumors
Administration of antibody that blocks CTLA-4 in
tumor-bearing mouse leads to tumor regression
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The PD-1 inhibitory pathway
• PD-1 recognizes two widely expressed
ligands (PD-L1, PD-L2)
• Knockout of PD-1 leads to autoimmune
disease (less severe than CTLA-4-KO)
• Role of PD-1 in T cell suppression in
chronic infections, tumors?
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T cell “exhaustion” in chronic viral infections
Memory T
cells:
Effector enhanced
Naïve CD8+ T cells antiviral
T cells responses
Acute infection:
clearance of virus
Virus
Chronic infection:
persistence of virus
Exhausted T cells: inability to
respond to virus
(expression of inhibitory
receptors, e.g. CTLA-4, PD-1)
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Actions of PD-1
• PD-1 attenuates TCR signaling in
responding T cells
• Limits harmful consequences of chronic
stimulation with persistent antigen (self,
tumors, chronic viral infections)
• Greater role in CD8 than in CD4 T cells
• Also expressed on follicular helper T
cells; function?
Checkpoint blockade for cancer immunotherapy
e.g. ipilimumab
Ribas A. N Engl J Med 2012;366:2517-2519.
Checkpoint blockade for cancer immunotherapy
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e.g. ipilimumab e.g. nivolumab, pembrolizumab
Ribas A. N Engl J Med 2012;366:2517-2519.
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Risks of blocking CTLA-4 or PD-1
• Blocking a mechanism of self-tolerance
leads to:
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Risks of blocking CTLA-4 or PD-1
• Blocking a mechanism of self-tolerance
leads to:
• Autoimmune reactions (a new cottage
industry for clinicians?)
– Colitis and dermatitis are common
– Vitiligo, Endocrinopathies, hepatitis less
common but described
– Severity of adverse effects has to be
balanced against potential for treating
serious cancers
– Less severe with anti-PD1 antibody
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Regulatory T cells
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 8th edition, 2014, Elsevier
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Properties of regulatory T cells
• Phenotype: CD4+, high IL-2 receptor
(CD25), low IL-7 receptor, Foxp3
transcription factor; other markers
• Essential features of stable Tregs:
– Foxp3 expression: requires demethylated non-
coding CNS2 sequence in promoter
– CD25 (IL-2Ra) expression: IL-2 is a necessary
survival factor
– CTLA-4 expression: required for suppressive
function of most Tregs
– (Inability to produce IL-2) Take home messages
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The significance of Foxp3+ Tregs
• Genetic evidence: Foxp3 mutations -->
autoimmune disease (IPEX); in mice,
disease can be corrected by providing
normal Foxp3+ cells
• Do defects in Foxp3+ Tregs or resistance
to Treg-mediated suppression contribute
to common autoimmune diseases?
– Inconsistent and variable data
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Mechanisms of action of Foxp3+ Tregs
• CTLA-4 on Tregs removes B7 on APCs,
reduces CD28 engagement and T cell
activation
– Genetic deletion of CTLA-4 in Foxp3+ cells
results in severe systemic autoimmunity and
lymphoproliferation
• Inhibitory cytokines produced by Tregs
(TGF-b, IL-10, others?) suppress immune
responses (DCs, Macs, T cells)
– IL-10 deletion in Foxp3+ cells results in colitis
– IL-10 is also produced by Foxp3- cells
• Consumption of IL-2
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Regulatory T cells
• Explosion of information about the
generation, properties, functions and
significance of these cells
• Will cellular therapy with ex vivo
expanded Treg become a reality?
• Therapeutic goal: induction or activation
of Treg in immune diseases
Take home messages
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The therapeutic potential of
regulatory T lymphocytes
• Cell transfer of autologous Tregs to
suppress immune responses
– Grow up patient’s Tregs ex vivo
– Ongoing clinical trials in graft rejection, T1D
show it is safe
– In one study of liver Tx, single infusion of
Tregs resulted in tolerance (withdrawal of
immunosuppression) in 7/10 patients (vs ~10%
historically)
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Functions of Interleukin-2: the dogma
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The unexpected biology of IL-2
• Interleukin-2 is the prototypic T cell
growth factor (TCGF), required for
initiating clonal expansion of T cells in
response to antigen
• BUT: knockout of IL-2 or the a or b chain
of the IL-2R results not in immune
deficiency but in systemic autoimmunity
and lymphoproliferation
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Dual roles of IL-2 in T cell responses
Surprising conclusion from knockout mice: the non-redundant
function of IL-2 is in controlling immune responses
Take home messages
R
due to IL-2R signalli
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PI3K–AKT and STA
Differential effects of IL-2 on
mTOR mTOR effector T cells in re
and IL-2, TReg cells —
Teff vs Treg
levels inhibit the P
Response Response way 43,44 — may be m
signalling (FIG. 2).
Reg
In vivo experime
ings. At doses of IL-
b TReg cells IL-2 international units
In vitro sensitivity to IL-2
100 100 are expanded compa
STAT5 phosphorylation (%)
STAT5 phosphorylation (%)
and they express high
80 80 are more suppressiv
60 60 clonal expansion nor
T cells or of NK cells
40 40
20 20
The inflammatory d
0 0 ing IL-2 or function
0.01 0.1 1 10 100 1,000 0.01 0.1 1 10 100 1,000
IL-2 IU ml–1 IL-2 IU ml–1
and the production
but the specific eff
In vivo sensitivity to IL-2
3 3 mouse strain. On th
Fold increase in the number
Fold increase in the number
nant phenotype is a
caused by erythrocy
of cells over baseline
of cells over baseline
2 2 C57BL/6 mice, the d
time, all Il2-knockou
bodies that are spec
1 1
self-antigens. Interes
duced even in germ
0 0 the colitis is largely
Baseline 5 days of Baseline 5 days of mice are made germ
low-dose IL-2 low-dose IL-2
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The n e w e ng l a n d j o u r na l of m e dic i n e
original article
Regulatory T-Cell Responses to Low-Dose
Interleukin-2 in HCV-Induced Vasculitis
David Saadoun, M.D., Ph.D., Michelle Rosenzwajg, M.D., Ph.D.,
Florence Joly, Ph.D., Adrien Six, Ph.D., Fabrice Carrat, M.D., Ph.D.,
Vincent Thibault, Pharm.D., Damien Sene, M.D., Ph.D.,
Patrice Cacoub, M.D., and David Klatzmann, M.D., Ph.D.
A BS T R AC T
Background
Patients with vasculitis induced by the hepatitis C virus (HCV) have reduced levels of From Université Pierre et M
regulatory T cells (Tregs). Resolution of HCV infection correlates with cure of vascu- Université Paris 06 (D.S., M.R.
D.K.); Centre National de la Rec
litis and the recovery of Treg levels. We reasoned that interleukin-2, a cytokine that entifique, Unité Mixte de Rech
promotes Treg survival and function, could be beneficial for patients with vasculitis (D.S., M.R., A.S., D.S., P.C., D.K
that is resistant to HCV therapy. Unité S959 (D.A., M.R., P.C.,
Unité 707 (F.C.); the Departme
nal Medicine, Reference Center f
Methods mune Diseases (D.S., P.C.), t
We investigated the safety and immunologic effects of the administration of low-dose Investigation Center in Biother
A.S., D.K.), and the Departme
Pathogenesis of autoimmunity 33
Therapy of immune disorders: rational approaches target 34
lymphocyte activation and subsequent inflammation
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Autoimmune diseases
• Experimental models are revealing
pathways of immune regulation
• But experimental animals are often inadequate
models of human diseases
• Improving technologies for human genetic
and phenotypic analyses are enabling
studies of patients
• Challenges:
– Defining which mechanisms of immune
tolerance fail in different autoimmune
diseases
– Using this knowledge to develop therapies
Take home messages
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The landscape of T cell activating and
inhibitory receptors: More to come?
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