IMMU3102 L16: Regulation of Immune Responses

Tolerance
-

No apparent response in animal Tolerance

o However, tolerance DOES involve a response, but it is an
abortive response that does NOT cause inflammation
Tolerance involves deleting T cells that recognise self-antigen in
lymph node
o Deleted T cells undergo rapid division for 2-3 days before they
die and they do NOT acquire effector function

What affects the type of immune response? What affects the role
of regulation?
-

Nature of antigen soluble vs particulate

Route of antigen intravenous, oral, subcutaneous, intramuscular,

intradermal
Type of APC DC
Affinity of receptors on activated lymphocytes self-reactive

receptors, foreign reactive

Prior antigen exposure affect role of regulation cant tolerise
individual cells once memory has been established dominant

suppression is needed
Age of individual not a direct affect but mediated via large
memory cell compartment

Different responses to different types of pathogens

-

External parasite drive Th2 responses

o Parasites such as worms
Intracellular pathogens drive Th1 responses
o Intracellular pathogens such as Mycobacteria

Regulation by Cytokines I
-

Cytokines can regulate each other cause interference

o E.g. IL-4 and IFN- are mutually antagonistic (Th1 produce IFN, Th2 produce IL-4)
However, Th1 and Th2 responses can be initiated
simultaneously in the same lymph node)
o Interference is most apparent at effector site:
Response to mycobacteria in lungs can suppress asthma
-

(an allergic response)

Cytokines are well adapted to work in non-lymphoid tissue sites
where lymphocytes are less closely packed than in lymph nodes
(cytokines needed to communicate over longer distance)

Regulation by cytokines II
-

TGF- suppress cell-mediated immunity important in mucosal

immunity (as TGF- drive IgA production)

IL-10 suppress antigen presentation in mice important in
preventing gut inflammation in mice, serves as B cell growth factor
in humans

Regulation of B cells by Antigen-Antibody complex and FcR

-

Immune complex (Ag-Ab) binding to FcR (receptor for antibodies)

prevent activation of nave B cell but does NOT affect memory B

cells
B cells can make antibodies that react with receptors on other B
cells anti-idiotypic antibodies downregulate responses thus B
cells can regulate each other

Effector T cells produce cytokines to regulate immune response

-

Th3 produce TGF- which suppress Th1 immunity

o However, TGF- induce Th17 cells that drive autoimmunity:
Brain inflammation Attack of myelin (EAE)
Attack of joints (collagen-induced arthritis)
o thus Th3 can induce autoimmunity via Th17 induction
Tr1 (type 1 Treg) produce IL-10 and IFN- (a Th1 cytokine)

o IL-10 provide negative feedback that limit size of Th1

response
o Th1 responses are accompanied by generation of Tr1 cells
Without Tr1 cells, Th1 cells can cause tissue damage
IFN- opposes effects of TGF- increase IL-12 at expense of IL-23
production stimulate differentiation of Th17 cells
o IFN- opposes the effect of TGF- in stimulating differentiation
of Th17 cells by increasing IL-12 production at the expense of
IL-23 production

Cells committed ONLY to regulation: Suppressor cells

-

Suppressor cells = Ts

Regulatory T cells (Tregs)

-

CD4+CD25+ T cells = T regs

T regs suppress proliferation of other T cells in vitro

Tregs are different to Th1, Th2, Th3, Th17, Tr1 which differentiate
from nave T cells after contact with antigen in lymph nodes and
spleen
o nTregs (natural regulatory T cells) differentiate as CD4+CD25-

in thymus
nTregs are selected after contact with self-peptide MHC in thymus
o nTregs are self-reactive but do NOT cause autoimmunity
nTregs PREVENT autoimmunity
o CD4+ T cells selected into nTreg population have an affinity
for self-antigen that is intermediate between that of cells
undergoing positive and negative selection.
o nTreg are activated by self-antigen after leaving the thymus.
Some nTreg may also cross-react with foreign antigen.

nTreg Phenotype
-

nTreg express IL-2R alpha chain (CD25) constitutively

nTreg express nuclear transcription factor, FoxP3.

o FoxP3 is required for nTreg function and is commonly used to

define nTreg subset.
o Unfortunately, FoxP3 is expressed inside the cell, so it cannot
be used to identify live nTreg cells
-

nTreg also express CTLA-4

nTreg express members of the TNFR family such as GITR at high

levels.

nTreg can make IL-10 and TGF- but whether these cytokines are
absolutely required for their effects in vivo remains controversial

nTreg use TGF- as a survival factor (i.e. they express TGF-

receptors at high levels)

All cell surface molecules and cytokines made by nTreg can be

expressed by activated T cells, so it is difficult to definitively identify
living nTreg in vivo, especially in humans.

Human nTreg (Lecturers research)

-

Human nTreg express low levels of CD127 (the alpha chain of the IL-

7 receptor) as a surface marker

CD127 is highly correlated with FoxP3 expression in nTreg cells.
o FoxP3 binds to the IL-7R alpha chain promoter and represses
transcription.

Use of cell surface markers to define human nTreg is necessary for

cell sorting before functional assays and in vitro expansion for
therapeutic purposes.

In vivo functions of nTreg

-

Mice with reduced nTreg numbers develop spontaneous immunoinflammatory disease that closely resembles immunoinflammatory
disease.
o Type of disease depends on genetic background of mice:
C57BL/6 mice develop inflammatory bowel disease,

 BALB/c develop gastritis, thyroiditis and oophoritis.

o Reconstitution with nTreg prevents disease in these animals.
-

Defects in IL-2, CTLA-4 and FoxP3 are associated with nTreg

deficiencies in mice

Defects in IL-2, CTLA-4 and FoxP3 are associated with development

of overwhelming multisystem lymphoproliferation and
autoimmunity.
o Mutations in FoxP3 cause a similar syndrome in humans
(immunodysregulation, polyendocrinopathy, enteropathy, X
linked (IPEX) syndrome)

Pathology associated with severe deficits in nTregs indicates the

importance of nTreg in maintaining normal balance within the
immune system

How do nTregs function?

-

nTreg reduce proliferation of other T cells in vitro by reducing

production of IL-2.
o This requires cell-cell contact

Human nTreg have cytolytic function.

nTreg can modulate adenosine/cAMP metabolism.

nTreg prevent activation of low affinity self-reactive T cells in vivo.

nTreg downregulate expression of co-stimulatory molecules by DCs

and B cells in vivo and in vitro thereby regulating threshold at
which T cells are activated by antigen This is probably how nTregs
prevent activation of self-reactive T cells.

Why are nTregs important?

-

Pathology of autoimmune disease in mice with defective nTreg

closely resembles that in humans thus studying nTregs in mice is
important

Patients with autoimmune diseases have reduced numbers of nTregs

or have nTregs that do not function properly.

Understanding what makes nTregs survive and function could give

us new ways of preventing and treating autoimmune diseases

Treg deficiency may be related to the hygiene hypothesis indicate

how Tregs work

Hygiene hypothesis and immune system

-

Allergic diseases and lifestyle factors:

o The incidence of allergic conditions is very high in Western
countries.
It is virtually nil in the third world.
Incidence of allergy is 25-40% of adult population.
10-15% of pre-schoolers suffer from asthma!
2% of pre-schoolers suffer from potentially lifethreatening allergy to peanuts.
o Epidemiological evidence links the incidence to lifestyle
changes as a result of high socio-economic status.
o It is believed that the link between high socio-economic status
and allergy is via hygiene i.e. lack of contact with microbes.

Allergists originally suggested that lack of Th1 stimulation by

microbes allows abnormally high Th2 responses that cause allergy.

Th1 vs Th2 as an explanation of high rates of allergy?

-

Parasitic infections stimulate Th2 responses but protect against

allergies

Th1 diseases (type 1 diabetes, Crohns disease) are also increasing

in the developed world

Autoimmune disease and lifestyle factors

Incidence of all autoimmune diseases is increasing at a similar rate

to that of allergic conditions in Western countries.
o These changes are too rapid to be attributable to genetic
changes thus lifestyle factors may contribute to increase in
autoimmune diseases

Autoimmune conditions now account for the third largest burden of

chronic disease after vascular disease and cancer (increasing
evidence that vascular disease is autoimmune, making
autoimmunity the major health problem in the western world!)

The incidence of autoimmune disease in the third world is very low.

This is not due to under-diagnosis - diabetes was first diagnosed in
ancient Greece and is easily diagnosed in the third world.

Hygiene Hypothesis
-

Possible cause of allergic and autoimmune disease

developed in the UK to explain rise of hay-fever, which had been

quite rare at the beginning of the 19th century
o Children were found to be less likely to develop hay-fever or
eczema if they had older siblings (having younger siblings had
no protective effect).

Original publication from epidemiologist David Strachan in 1989

concluded:
o Allergic diseases were prevented by infection in early
childhood, transmitted by unhygienic contact with older
siblings, or acquired prenatally from a mother infected by
contact with her older children.

Evidence for Hygiene Hypothesis

-

Migration studies show that birthplace is very important rates of

allergic and autoimmune disease increase in children born after
migration to high socio-economic status countries

Early childcare is protective in countries with significant recent

migration from low socioeconomic status countries.

Lack of contact with microbes is important, rather than hygiene

itself (i.e. hand-washing, food preparation practices etc.)

Immunological disease and the hygiene hypothesis

-

Exact nature of microbes is unknown but most likely to be gut

flora

Ten times more bacteria in human large bowel than cells in the body
(1013-1014).
o At least 1000 species most not currently cultivatable

More than 70% of the human immune system is focused on the

bowel (i.e. 70% of immune responses occur in the bowl)

Microbes first colonise during birth

Changes in health practices (e.g. birth practices, breastfeeding etc.)

and hygiene may have changed human microbiome reduced
transmission in each successive generation) correlate with
increased incidence of immuno-inflammatory disease

Does hygiene affect the number or function of nTreg?

-

Knowing which immunological pathway is involved in the hygienerelated immunological problems find ways to replace
environmental signals required to prevent allergy and autoimmunity

Summary
-

Regulation occurs at all levels in the immune response.

For every positive signal, there is a negative pathway to keep the

stimulation under control.

T and B cells can directly regulate themselves AND each other, as

well as all the other cells involved in immune responses

Mechanisms of regulation: Ab, cytokines, cell-surface contact,

and the targets are lymphocytes, dendritic cells, and effector cells
such as macrophages

nTreg compartment is the most potent in preventing autoimmune

disease
o Treg is the most potent regulatory compartment involved in
preventing autoimmune disease