IMMU3102 L17: Regulation of Immune Response II

o B cells generated in Bone marrow

o T cells generated in Thymus
Recall: Developing Immune System
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Generation of diversity via Combinatorial Diversity and Junctional

Diversity:
o Combinatorial diversity:
due to random recombination f V, D, J segments
limited by number of V, D, J segments
o Junctional Diversity:
Generates MORE diversity by random ADDITION and

SUBTRACTION of nucleotides at V-D-J or V-J junctions

Thus almost limitless possibilities

Recall: Developing Immune System

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Immune system is developed via generation of diversity of

receptors:
o Two types of diversity: Combinatorial diversity and Junctional
diversity:
Combinatorial diversity is due to random recombination
of V, D, J segments limited by number of V, D, J

segments limit diversity

Junctional diversity is due to random ADDITION and
SUBTRACTION of nucleotides at V-D-J or V-J junctions
thus almost limitless diversity

B cells are generated in Bone marrow

T cells are generated in Thymus

Random recombination event generate equal of amounts of

receptors that recognise self-antigens and receptors that dont
recognise self-antigens
o Receptors that dont recognise self-antigens selected

o Receptors that do recognise self-antigens deleted or

regulated (i.e. become Tregs)

Factors determining Autoimmunity

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Genetic factors: failure of tolerance, unregulated lymphocyte

activation
Environmental factors: infections, smoking

Factors lead to T and B cells responding to self-antigens

Regulating immunity
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Little bit of inflammation is good

Too much inflammation might kill you

Summary
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Initiation of primary immune response is restricted to DC

Multiple signals are required to activate a nave T cell to become an

activated effector T cell

Inappropriate immune response can lead to tissue damage,

inflammation, systemic cytokine storms and autoimmune disease

Stages of Adaptive Immune Response that need to be regulated:

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Peripheral sites
T cell activation and differentiation
Within Germinal centres

Interfering with immune regulation can have therapeutic

benefits:
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Using Ipilimumab:

o Monoclonal antibody used to treat melanoma

o Bind to CTLA-4 on both effector T cells and Tregs
o Cause antitumor activity

How is immune system regulated?

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Regulating Complement activation:

o Recall (week 9): mammalian cells express regulatory proteins
that inhibit complement activation prevent complementmediated damage to host cell
o C1 inhibitor inhibit complement activation early (at the C1
stage of activation, classical and lectin pathway)
o MCP (CD46), Factor I inactivate C3b fragments
o Factor H Binds C3b and displaces Bb inhibit Alternative
pathway
o C4 binding protein disrupts C3 convertase
o DAF disrupt binding of Bb to C3b, bind C4b to C2a inhibit
Alternative and Classical pathway
o CD59 block C9 binding prevent formation of MAC
o Recall: complement system is activated on microbial surfaces
and NOT on normal host cells because microbes lack these
regulatory proteins

Regulation by cytokines:
o Cytokines regulating each other:
IL-4 and IFN- are mutually antagonistic (regulate each
other)
o IL-10 supress antigen presentation prevent development
of Adaptive Immune Response
o TGF- suppress cell-mediated immunity

Regulation by cytokine-producing effector T cells:

o nTregs express CD25 (IL-2R alpha chain) and FoxP3 (nuclear
transcription factor) constitutively
o nTregs outcompete T cells for IL-2 prevent T cell activation
o nTregs express CTLA-4
o nTregs can produce IL-10 and TGF- (immunosuppressive
cytokines)

Both Innate and Adaptive responses need to be regulated

Feedback mechanisms that regulate Innate Immunity
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IL-1 receptor antagonist (IL-1RA)

o produced by Phagocytes
o IL-1RA is similar to IL-1 but NOT biologically active (IL-1 is
proinflammatory cytokine)
o IL-1RA bind to same receptor as IL-1 function as competitive
IL-1 inhibitor
o Synthesis of IL-1RA is induced by same stimuli that induce IL-1
production
o IL-1RA is required to prevent inflammatory disease of joints
and other tissue
o Recombinant IL-1RA is developed as a drug

Regulating NK cell recognition of infected cells:

o NK cell inhibitory receptors recognise MHC class I molecules
avoid killing normal cells

Negative regulatory signalling pathways:

o Blocking activating signals produced by PRRs
o Block inflammatory cytokines
o Inhibitors of JAK-STAT signalling pathway (linked to cytokine
receptors): SOCS (suppressors of cytokine signalling proteins)
o Expression of SOCS protein via TLR signalling in macrophages
and DC
o Activation of SOCS protein limit response of cells to cytokines

B cells and immune regulation

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B cells differentiated into ASCs (Antibody Secreting Cells)

Bregs secrete IL-10 regulate immune response

o IL-10 protect against autoimmunity e.g. Multiple sclerosis
o IL-10 prevent inflammation e.g. Atherosclerosis, Asthma
o IL- 10 enhance transplantation success lower chance of
GVHD (Graft Versus Host Disease)

o IL-10 inhibit anti-tumour immunity thus promote tumour

growth
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Thus b cells have dual immunological role produce Antibodies to

protect us, secrete IL-10 to maintain peripheral tolerance

B cells create immunity against extracellular pathogens

antibodies bind to pathogens

Antibodies secreted by B cells form the Humoral immune response

B cells and autoimmunity

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B cells are required to prevent autoimmunity

Bregs also required to treat autoimmunity

How are Bregs activated and how do they suppress?

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DC activate Th cell:
o MHC/peptide + TCR
Th cell activate B cells:
o CD40 (on B cell) + CD40L (on Th cell)

TLR stimulation also required to activate B cell

BCR activation also required

Activated B cell produce IL-10 suppress autoimmune diseases

no activated Th1 and Th17

Bregs and Asthma

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Bregs can suppress asthma:

o Bregs known as B10s produce IL-10 reduce inflammation
o B10s suppress CD4+ T cells reduce IFN- and TNF-
production
o B10s reduce DC antigen presentation Reduce Th2 activation

Bregs and Cancer

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Bregs and its Cytokines:

o IL-10 suppress CD8+ T cell ability to secrete IFN- (proinflammatory cytokine) thus inhibit cell-mediated immunity
o TGF- (anti-inflammatory suppressive cytokine) control
CD4+ T cell differentiation to Tregs

Tumours use Bregs suppress CD8+ effector function and to

activation of Tregs lead to progression of tumour

Thus Bregs can be used by tumour to evade immune system

grow uncontrollably

Inhibit Bregs inhibit progression of cancer

o Ipilimumab targets Tregs

Innate Lymphoid Cells (ILCs) and Immune Regulation

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ILCs originate from lymphoid organs but LACK recombined Antigen

receptors

ILC subsets are represented at mucosal surfaces antimicrobial

functions at those surfaces

Group 3 ILCs prevent exuberant immune response to commensal

bacteria (helpful bacteria)

Type 2 ILCs in Skin and immune regulation

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Skin ILCs = Dermal ILC2 cells

ILC2 produce IL-13 in steady-state conditions
ILC2 promote eosinophil influx and cutaneous inflammation
ILC2 exert regulatory function on other dermal population esp.
Dermal Mast cells

Mast cells and immune regulation

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Regulatory Mast Cells = MCregs

MCregs are activated by IL-9 secreted by Tregs

MCregs are activated by Vitamin D from sunlight

MCregs are activated by cytokines: SCF, IL-3, IL-4, IL-33

How do MCregs suppress?

o MCregs produce anti-inflammatory cytokines: IL-10 and IL-13

What
o
o
o
o
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diseases are effected by MCregs?

Dermatitis
Transplantation
Allergy/asthma
Autoimmunity
Cancer

Regulatory Mast Cells and IL-10

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MCregs IL-10 effector functions:

o Reduce inflammation
o Prevent transplantation rejection via induction of Tregs
o Promote tumour growth (IL-10 is an anti-tumour immune
suppressive cytokine)

MCregs produce angiogenic factors, VEGF (vascular endothelial

growth factor)
o VEGF is produced for angiogenesis
o VEGF promote creation of new blood vessels that helps feed
cancers

Thus Mast Cells can be used to promote cancer growth

MCregs supress germinal centre formation by inhibiting TFH

Mast cells thus have dual immunological role

Mast cell regulation with IL-10

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IL-10 from Mast cells:

o reduce Fc receptors on Mast cells prevent binding of IgE
prevent degranulation

o supress DC maturation and activation DC cannot activate T

cells
o promote production of Tregs (Tregs then produce more IL-10)
o downregulate co-stimulatory molecule expression by DCs
reduce T cell proliferation and cytokine production suppress
development of effector T cells
o reduce TNF and IL-6 production of Keratinocytes in Periphery
suppress proinflammatory signals in skin less leukocyte
recruitment

Germinal Cell and Immune Regulation

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inside germinal centre:

o dark zone mutation occurring in BCR increase affinity
insertion of mutation can lead to production of autoantibodies
o light zone BCR tested
o Extrafollicular zone where isotype class switching occur
Where IgM are produced then switched (depends on
type of Th cell present)

IL-10 produced by Mast cells suppress germinal centre formation

T follicular helper cells = TFH

Regulating TFH cell differentiation:

o Cell-intrinsic factors: Transcription factors
o Cell-extrinsic factors: cytokines

T Follicular Regulatory Cells and Germinal Centre Regulation

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T Follicular Regulatory Cells = TFR

Initially:
o Th cell migrate from Extrafollicular zone into germinal centre,
form germinal centre
o TFH secrete IL-21 help produce high affinity class switched
antibody

Treg at extrafollicular zone becomes TFR migrate into germinal

centre suppress development of high affinity class switched
antibody production

Macrophages and Immune Regulation

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Microbes not only get rid of pathogens but ALSO promote

development of cancer
Classically-activated macrophages: M1
o M1 ingest pathogen and process antigen via MHC-class II
processing pathway
o M1 reside in the periphery and encourages inflammation

CD4+ T cells recognise antigens presented by macrophages on MHC

class II molecules

Macrophages receive help from CD4 T cells in two ways:

o CD40-CD40L interaction
o IFN- receptors on Macrophages bind to IFN- produced by
Th1 cells
o Thus without CD4 T cells (esp. Th1), you cannot effectively
eradicate intracellular pathogens

Activated macrophages carry out effector functions: Eliminate

Intracellular pathogens
o Secretion of cytokines: TNF, IL-1, IL-12
o Secretion of chemokines
o Increase expression of MHC molecules
o Upregulation of co-stimulators
o Kill phagocytosed bacteria

Macrophage Responses and its role in Cell-mediated immunity

(i.e. role in eliminating intracellular pathogens)

Production of ROS and NO (Reactive Oxygen Species, Nitric Oxide)

enhance killing of phagocytosed in phagolysosomes (i.e.
intracellular pathogens)

Secretion of TNF, IL-1, IL-12, chemokine recruit leukocytes, cause

Th1 differentiation, suppress Th2 differentiation

Increase expression of MHC molecule and co-stimulators increase

T cell activation and help

M2 Macrophages and Immune Regulation

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Are not classically-activated but ALTERNATIVELY-activated

o Alternatively activated macrophages = M2 macrophages

M2 are induced by IL-4, IL-10, IL-13

o These cytokines are produced by Th2, Leukocytes (eosinophil,
mast cell, basophil, tumour cells)
Thus tumour cells can activate M2

M2 is involved in tissue repair, remodelling and fibrosis (during

infections and allergic diseases)

M2 secrete IL-10 and TGF- anti-inflammatory effects

o TGF- cause new blood vessels to form can support tumour
cells
o Thus cytokines produced by tumour cells can activate M2
promote tumour cell growth

M2 secrete IL-4 suppress classically-activated macrophages (M1)

o IL-4 stimulate production of IL-10 and TGF- secreted inhibit
Th1 development cannot activate M1

Thus M2 play a role in immune regulation

o However, when found in Cancer known as Tumourassociated Macrophages

M1 and M2 summary
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M1:
o Produce pro-inflammatory cytokines
o Remove intracellular pathogens with help of Th1 cells
o M1 involved in removing Cancers
Cancers produce cytokines which prevent development
of M1 macrophages
M2:
o Promote angiogenesis, remodelling get new blood supply
help tumours grow
o Secrete TGF- and IL-10 inactivate DC, inhibit Th1
development prevent activation of M1
o Activate Tregs
o Inhibit CTL development
o Thus M2 plays a regulatory role yet in the presence of cancer,
it promotes cancer growth by inhibiting immune response

Excessive M1 activation
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Excessive M1 too much inflammation Cause Diseases:

o Arthritis
o Diabetes

Excessive M2 activation
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Cause:
o Granuloma formation (during chronic phase of mycobacteria
and tuberculosis infection)
o Promote cancer development (breast cancer, skin cancer)
o Fibrosis
o Asthma (airway inflammation)

Summary

Stages of Adaptive immune response that needs to be regulated:

o Regulation at Peripheral sites (where antigen is encountered):
Regulating Mast cells, Macrophages, Neutrophils, ILCs,
DC, Tregs
o Regulating Antigen presentation to nave T cells:
Regulating DCs, Bregs, Tregs, mast cells
o Regulating T cell differentiation:
Regulating Mast cells, Bregs, Tregs
o Regulating Germinal centres:
Regulating Mast cells, T follicular regulatory cells