Antigen Presentation &

MHC Molecules

CYTOKINES

1
 Functions of T lymphocytes

Primary defense against intracellular microbes (antigens)

Activation of other cells (phagocytes, B cells)
Killing of infected cells (i.e., CTL)

Key features:
Mediated by interactions with other cells
Allows surface molecules, cytokines to act at short range
(enhances specificity)
Different classes of T lymphocytes are most effective
against different types of microbes (antigens)
Phagocytosed (extracellular) microbes/antigens: helper T cells
stimulate antibody production, macrophages
Cytoplasmic (endogenous microbes/antigens): CTLs kill infected
cells and eliminate reservoirs of infection

2
 The challenge for T lymphocytes

Very few lymphocytes in the body are specific for

any one microbe (or antigen)
Specificity and diversity of antigen receptors: the immune
system recognizes and distinguishes between 107 - 109
antigens
The frequency of antigen responsive lymphocytes is
typically 1 in 105 to 106

Lymphocytes must be able to locate microbes

(antigens) that enter anywhere in the body

Lymphocytes must respond to each microbe

(antigens) in ways that are best able to eradicate that
microbe (antigen). 3
APCs are Required to Present Antigenic Peptide
Fragments to T cells

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Induction of immune responses

Antigens (microbes) enter through interfaces with

external environment
(portals of entry: skin, GI tract, respiratory tract)

Antigens are transported to peripheral lymphoid

organs (lymph nodes, spleen) by professional APCs
(Dendritic Cells) or in soluble form

Nave lymphocytes that recirculate through the

same organs locate the antigens and are activated

5
Mannose receptors, and expression of
C-type Lectin receptors CCR7, and upregulation of
and Toll-like receptors MHC I & II, and coreceptors

6
 Why are dendritic cells the most efficient APCs for
initiating immune responses
Location
At sites of microbe entry (epithelia), tissues
Express receptors (CCR1,2, and 5) that recognize
inflammatory cytokines
Receptors for capturing microbes & antigens
Such as mannose and C-type lectin receptors and Toll-
like receptors (TLRs) which function as pattern
recognition receptors and have very active endocytic
machinery
Migration to T cell zones of lymphoid organs
Role of CCR7 (ligands are Mip-3b and SLC)
Co-localize with nave T cells
Maturation during migration
Increase levels of MHC molecules, induce costimulators
(B7 molecules, CD40) and decrease endocytic capacity
Conversion from cells for antigen capture into cells for
antigen presentation and T cell activation 7
 Dendritic cell subsets
Immature dendritic cells capture antigens from
sites of entry

Mature (activated) DCs present antigen to and

activate nave T cells in lymphoid organs

DC1, DC2 subsets may stimulate differentiation to

different T cell subsets (Th1, Th2)

Some DCs (i.e., immature) may induce tolerance

Problems:
Lack of definitive markers
8
Most studies based on culture derived DCs
Maturation of Dendritic Cells

MHC

9
Functions of Different Antigen Presenting Cells

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
Is T cell recognition of virus genetically controlled?

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Self MHC Restriction of T cells

The genes that differ between strains A and B and control T cell
recognition were found to map to a locus called the MHC 12
Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 What is the MHC?
A genetic locus discovered on the basis of
transplantation
Different individuals express products of different MHC
alleles and reject grafts from one another
Human MHC: HLA (human leukocyte antigens)
Mouse MHC:H2
The peptide display molecules of the immune
system
Different alleles of MHC molecules display distinct
but overlapping sets of peptides
Determines which protein antigens are recognized in
different individuals
MHC molecules determine how antigens in different
cellular compartments are recognized by different
classes of T cells 13
 MHC (major histocompatibility complex)-restricted
antigen recognition by T cells

A T lymphocyte recognizes an antigen on an antigen-

presenting cell (APC) that also expresses an MHC allele
that the T cell sees as self
First indication that T cells recognize complex of protein
(peptide) antigen bound to MHC molecules

Antigen receptors of T cells have dual specificities:

1. For peptide antigen (responsible for specificity of immune
response), and
2. For polymorphic residues of self MHC molecules (responsible
for MHC restriction)

T cells learn self MHC restriction during maturation in the

thymus: only T cells that see MHC molecules in the thymus
(can only be self MHC molecules) are selected to survive 14
15
16
 Features of Class I and Class II MHC Molecules
Feature Class I MHC Class II MHC

Polypeptide chain a (44-47 kD) a (32-34 kD)

b2-microglobulin (12kD) b (29-32 kD)

Polymorphic residues a1 and a2 domains a1 and b1 domains

Coreceptor a3 domain binds CD8 b2 domain binds CD4

Source of peptide Cytosol (biosynthetic pathway) Endocytic/lysosomal

pathway

Peptides bound 8-11 residues 10-30 residues

Nomenclature
Human HLA-A, -B, or C HLA-DR, -DQ, -DP
Mouse H-2K, H-2D, H-2L I-A, I-E

Types of APCs All nucleated Cells DCs, Macrophages,

17
B cells
 Structure of Class I MHC Molecule

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003

19
 Some important properties of MHC molecules
MHC molecules are the immune systems mechanism
for displaying peptide antigens to T lymphocytes
Highly polymorphic genes: large number of alleles in the
population
Co-dominantly expressed: each cell has six class I
molecules (3 from each parent) and 10-20 class II molecules
(3 from each parent + some hybrid molecules)
Class I MHC molecules are expressed on all nucleated cells
Class II MHC molecules are expressed on few cells types
(specialized APCs, e.g. dendritic cells; B lymphocytes,
macrophages)
Stable expression of MHC molecules on cell surfaces
requires the peptide cargo
MHC molecules present foreign and self peptides
Expression of Class II MHC molecules, in particular, is up-
regulated by activation of the innate immune response (IFNs,20
etc.)
Enhancement of Class II MHC Expression by IFNg

Similar effects
on class I MHC and
the activation of
CD8 T cells.
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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
22
Polymorphic residues of MHC molecules

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
Alleles of MHC Molecules are Codominantly Expressed

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Presentation of Extracellular and Cytosolic Antigens

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Antigen processing

Conversion of native antigen (globular protein)

into peptides capable of binding to MHC
molecules

Occurs in same cellular compartments as

synthesis and assembly of MHC molecules
Determines which source of antigen generates
peptides that are displayed by class I or class
II MHC
Uses cellular organelles that serve basic
housekeeping functions in most cells
26
Pathways of Intracellular Processing of Protein Antigens

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
The class II MHC pathway of processing of
internalized vesicular protein antigens

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Class II MHC pathway of presentation of
vesicular peptide antigens

Proteins ingested into endosomes/lysosomes are

processed and presented in association with class II
molecules
Most vesicular peptides are derived from extracellular
proteins
CD4 binds to class II MHC; therefore, CD4+ T cells
recognize class II-displayed peptides (only some cell
types express class II MHC)
CD4+ T cells are helper cells that activate B
lymphocytes and macrophages
Antibodies and macrophages attack and destroy
extracellular microbes
29
 The class I MHC pathway of processing
of endogenous cytosolic protein antigens

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Class I MHC pathway of presentation of cytosolic
peptide antigens

Cytosolic proteins are processed into peptides and

presented in association with class I molecules
Most cytosolic peptides are derived from
endogenous (e.g. viral, tumor) proteins; some may be
from phagocytosed microbes, proteins enter cytosol
CD8 binds to class I MHC; therefore, CD8+ T cells
recognize class I-displayed peptides
CD8+ T cells give rise to cytotoxic T lymphocytes
(CTLs) that kill other cells that harbor infections or
are transformed (all nucleated cells express class I
MHC)
CTLs destroy cells infected with intracellular
microbes and eradicate reservoirs of infection
31
How class I- and class II-associated antigen presentation
influence the nature of the host T cell response

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Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003
 Significance of MHC-associated antigen presentation
MHC molecules display foreign and self peptides from
the extracellular and intracellular environment
T cells survey the body for foreign (microbial) peptides
Different classes of MHC molecules present cytosolic
(endogenous) and vesicular (ingested) peptides
Helper T cells and CTLs respond to the microbes (antigens)
that each is best able to combat
T cell receptors only recognize MHC-peptide
complexes, and MHC molecules are cell surface
proteins
T cells interact with other cells and not with cell-free antigens
Only peptides bind to MHC molecules
T cells recognize only proteins (natural source of peptides)
Few peptides are presented even from complex
proteins 33
Immunodominance: few peptides bind to any MHC molecule
 Immunodominance of Peptide Epitopes

Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003

Determinant Selection - MHC alleles select best binding

peptides and thereby select which determinants will be
immunogenic in an individual. 34
 The problem for CD8 T cells
Viruses and tumors may be present in any
nucleated cells; therefore, the immune system has
to be able to generate CTL responses (class I-
restricted) to any nucleated cell

Only some APCs, particularly DCs, are able to

initiate the responses of nave T cells

How are antigens from virus-infected or neoplastic

non-APC cell types transferred to APCs?

35
Cross-presentation of antigens to CD8+ T cells

Abbas & Lichtman. Cellular and Molecular Immunology, 5th ed. W. B. Saunders 2003

-explains how infections of non-professional APCs can lead

to the initiation of a CD8 T cell response
36
 Functions of APCs
Capture antigens and take them to the correct
place
To peripheral lymphoid organs, through where nave
lymphocytes circulate

Display antigens in a form that can be recognized

by specific lymphocytes
For T cells: MHC-associated peptides (cytosolic peptides
to class I, vesicular peptides to class II)
For B cells: native antigens; APCs include macrophages,
follicular dendritic cells in germinal centers

Provide second signals for T cell activation

Costimulators and cytokines induced by microbes
(antigens); ensure that T cells respond best to microbial
37
antigens
 CYTOKINES

Cytokines are a group of low molecular weight regulatory

proteins secreted by leukocytes as well as a variety of other
cells in the body in response to stimuli. Membrane-bound
forms of some of the secreted cytokines are known as well.

Cytokines regulate the intensity and duration of the immune

response by stimulating or inhibiting the activation,
proliferation, and/or differentiation of various cells, and by
regulating the secretion of antibodies or other cytokines or
mediators.

Almost ALL cells secrete at least a few cytokines.

38
 MODES OF CYTOKINE TRAVEL

INTRACRINE
JUXTACRINE

AUTOCRINE PARACRINE
ENDOCRINE

BLOOD VESSEL

39
from Bafico & Aaronson, Cancer Med, 2002
 3 MAJOR ROUTES OF CYTOKINE TRAVEL

Autocrine e.g. IL-2 on

T cells - clonal
expansion

STIMULUS
Paracrine
e.g. T cell
cytokines act on
Endocrine B cells - isotype
switching
e.g., IL-1 &
fever

Blood or lymph vessel

40
Cytokines differ from growth factors in the structure of the receptor.

IGF-1R IL-6R
Lacks Tyr kinase domain
Has Tyr kinase domain
JAK (a kinase) docks instead

41
 MOST CYTOKINE RECEPTORS ARE IN 2 CLASSES

CLASS I CYTOKINE RECEPTORS

Most of the cytokine-binding receptors that function in the immune and

hematopoietic systems belong to this receptor family. There are conserved
amino acid sequence motifs in the extracellular domain - 4 positionally
conserved Cys residues (CCCC) and a conserved sequence of Trp-Ser-X-
Trp-Ser (WSXWS) where X is a nonconserved amino acid. The receptors
consist of 2 polypeptide chains: a cytokine-specific subunit and a signal-
transducing subunit, which is usually not specific for the cytokine. The
signal transducing subunit is required for high affinity binding of the
cytokine.

CLASS II CYTOKINE RECEPTORS

These receptors possess the conserved cysteine motifs, but lack the
WSXWS motif present in class I cytokine receptors.

42
 CYTOKINES BY FUNCTION

CYTOKINES THAT MEDIATE AND REGULATE INNATE IMMUNITY

Type I interferons
Tumor necrosis factor-a
Interleukins 1, 6, 10, 12, 15, 18, and more
Chemokinesa special subclass

CYTOKINES THAT MEDIATE AND REGULATE SPECIFIC IMMUNITY

Interleukins 2, 4, 5, 13, 16, 17, and more
Interferon-g (Type II interferon)
Lymphotoxin-a (Tumor necrosis factor-b)

CYTOKINES THAT STIMULATE HEMATOPOIESIS

c-kit ligand
Interleukins 3, 7, 9, 11, more
Colony stimulating factors M-CSF, G-CSF, GM-CSF, EPO, TPO

Overlapping functions are common.

43
 IL-1 FAMILY

Potent inflammatory effects, especially IL-1a & IL-1b. IL-18 induces IF-g, bridging
Inflammatory & innate/adaptive immune responses. IL-1 induces TNF, IL-6 as well as
Inflammatory mediators from various cells, notably cells at the interface of the
environment. IL-1 self-regulates through induction of IL-1Ra & IL-10. IL-1 is an
endogenous pyrogen (fever inducer). IL-33 induces TH2 cytokine synthesis, polarizing
to TH2 (Ab) responses.

IL-1 Family Receptors

Two receptors (RI and RII), both members of Ig gene superfamily.

IL-1RI found on T and B cells, fibroblasts, keratinocytes, endothelial cells,
chondrocytes, hepatocytes, and synovial lining cells; has a MW of 80 kDa.
IL-1RII found on B cells, neutrophils, & bone-marrow cells; has a MW of 60 kDa.

Difference in MW arises from IL-1RII having shorter cytosolic region.

Some evidence that 2 receptors mediate different biological events even though they
both bind IL-1 & IL-18.
A 3rd non-signaling receptor is found as well, possibly with regulatory function.
IL-33 signals only through RII.

RI 44
IL-1 IL-18

Signal (bacterial, etc.) to monocytes, epithelium, etc. Signal (bacterial, etc.) to monocytes

IL-1 IL-18

IL-6 TNF-ab IF-g

MHC Class I & II, IL-18

Req. for Ig class Dendritic cell maturation
switching
Enhanced Ag pres. to T cells
Very enhanced Ag pres. to T cells
Higher avidity Ab
response T cell effector functions: T cell effector functions:
TH1 TH1
TH2----B cells make Ab TH2----B cells make Ab
TH17 TH17
CTL CTL

(Monocytes can be macrophages in blood or tissue.)

45
 ENDOCRINE IL-1 PRODUCES FEVER

a virus
(not to scale)

IL-1
THIS WAY TO
THE BRAIN!

Blood vessel

46
 IL-2-LIKE CYTOKINES
The common g subunit associates with each
specific cytokine receptor to form the high
affinity cytokine receptor. The a subunit has
a very short cytoplasmic tail; it probably does not
signal.

IL-2
secreted primarily by T helper lymphocytes activated by stimulation
with T cell mitogens, or by interaction of T cell receptor complex
with Ag/MHC complexes on surfaces of antigen-presenting
cells. The response of TH cells to activation is induction of IL-2 and
high affinity receptors for IL-2 and, subsequently, clonal
expansion of antigen-specific T cells. Here IL-2 is an autocrine
factor, driving the expansion of the Ag-specific T cells.

IL-2 also acts as a paracrine factor, influencing the activity of other

cells, both within & outside of the immune system. B cells and
natural killer (NK) cells respond, when properly activated, to IL-2.

The so-called lymphokine activated killer,or LAK cells, appear to be

derived from NK cells under the influence of IL-2.

47
AUTOCRINE IL-2
DRIVES CLONAL IL-2
EXPANSION OF Ag-specific
T cell
T CELLS stimulus

IL-4

PARACRINE IL-4 B cell

DRIVES CLONAL
EXPANSION OF
B CELLS

AUTOCRINE OR PARACRINE
IL-4 OR IL-13 DRIVES
IgG1 IgE
Ig CLASS SWITCHING
TO IgE

48
 IL-4
Stimulates production of Ab-producing B cells, leading to the production of Ig
& class-switching to IgE. It also promotes CD8+ cell growth and TH2 cell differentiation.
On macrophages, IL-4 induces MHC class II expression, but inhibits production of pro-
inflammatory cytokines (IL-1, TNFa).

Induction of IL-4 secretion is Ag-specific and MHC restricted. Antigenic stimulation also results
in increased responsiveness of TH cells to IL-4. Ag-induced proliferation of TH cells is inhibited by anti-IL-4.
Thus, IL-4 mediates the proliferation of TH cells by an Ag-induced autocrine mechanism.
During T cell--B cell interactions involving soluble protein antigens, IL-4 and not IL-2
is the critical cytokine for activating resting B cells and inducing proliferation of the TH cells.

IL-9
Up-regulates TH1 responses by inhibiting T cell apoptosis.

IL-9 is preferentially produced by TH2 cells and is active on various cell types such as T and B cells, mast cells
and hemopoietic progenitors.

IL-13
Has structural and functional similarities to IL-4 and promotes B cell differentiation.
Also promotes Ig class switching to IgE.
IL-15
Shares several activities with IL-2 and is produced by epithelial cells and monocytes.
IL-15 induces T cell proliferation, enhances NK cell cytotoxicity and stimulates B cells to
proliferate and secrete Ig. 49
 IL-7

Is a T cell growth and activation factor, and a macrophage activating factor.

Interleukin-7 exerts pleiotropic effects on the immune system; it affects pre-B cells,
thymocytes, mature T cells, lymphokine activated killer cells (LAK), monocytes, and
macrophages.

IL-7 stimulates the proliferation of pre-B cells harvested from bone marrow. IL-7 activates these
cells in vitro.

IL-7 restores V(D)J rearrangement to developing T cells outside of the thymic environment.

Normal maintenance and proliferation of thymic progenitor cells requires the presence of IL-7.
In combination with phorbol 12-myristate 13-acetate, IL-7 drives the activation of resting
CD4+ and CD8+ T-cells along a pathway that is independent of IL-2.

IL-7 maintains nave CD4+ T-cells in vitro for up to 15 days, which suggests that it is a survival
factor for these cells.

Peripheral blood monocytes are activated by IL-7: lyse tumor cells, secrete IL-6, IL-1a, IL-1b,
and TNF-a.

Cyclosporine A (CsA), an immunosuppressive drug, inhibits the transcription of many cytokines

(for example, IL-2 and IL-4), but does not affect IL-7.
50
 IL-3, IL-5, GM-CSF
IL-3

Produced by activated T cells. Stimulates the proliferation of all hematopoietic precursors.

Stimulates differentiation of osteoclasts from bone marrow.

IL-3 IL-5

Is chiefly a growth and activation factor for eosinophils and B cells.

Eosinophils in particular are noted for their contribution to late phase allergic-type disorders. Eosinophils
make up less than 10% of the circulating leukocyte population, yet are extremely important in the
inflammatory response to allergic and parasitic challenges.
Eos are the cells of the Ab-dependent cell-mediated cytotoxicity responses (ADCC-- tumors, parasites).
Cells known to express IL-5 include eosinophils, NK cells, CD8+ T cells, mast cells, CD4+ T cells, g T
cells, and IL-1b-activated endothelial cells. IL-5 is best known for its activity on B cells and eosinophils.
IL-5 IL-5 appears to induce the differentiation of activated conventional B-2 cells into Ig-secreting cells.
In addition, it induces the growth of B-1 progenitors as well as IgM production by B-1 cells.
In mice, IL-5 promotes production of IgA, IgE and IgG1.

GM-CSF

Is chiefly a differentiation factor for myeloid cells -- neutrophils, macrophages. G = granulocyte.

GM-CSF 51
 IL-6 IL-6-LIKE CYTOKINES
IL-11 IL-6
Stimulates many types of cells. Endocrine response by liver to it results in
the acute phase response. Required for Ig class switching and for B cells
to differentiate into plasma cells. Endogenous pyrogen.

While a number of Interleukins such as IL-1 and IL-10 are seemingly pleiotrophic
in their effects, IL-6 may be considered the prototypic pleiotrophic cytokine.
a This is reflected in the variety of names originally assigned to IL-6 based on
function, including Interferon b2, IL-1-inducible 26 kD Protein, Hepatocyte
gp130 leptin Stimulating Factor, Cytotoxic T-cell Differentiation Factor, B cell Differentiation
Factor (BCDF) and/or B cell Stimulatory Factor 2. Once all the activities
associated with the names for IL-6 became connected with one common
CNTF gene, the name IL-6 was proposed. A number of cytokines make up an IL-6 cytokine
CT-1 family. Membership in this family is based on a helical cytokine structure and
receptor subunit makeup.
LIF Just about every cell type seems to express IL-6; expression of the others more
NNT-1 limited. IL-6 production is correlated with 10 cell activation but others with 20.
OSM
IL-6 has been described as both a pro-inflammatory and anti-inflammatory
IL-31 molecule, a modulator of bone resorption, a promoter of hematopoiesis, and
an inducer of plasma cell development. IL-6 also has been shown to influence
IL-4 production. It has been suggested that antigen-driven, APC-derived IL-6
may influence naive CD4+ T cells to produce IL-4 and express the IL-4
receptor. Thus, given the close approximation of APC and T cell, transient
APC-derived IL-4 could initiate a T cell autocrine loop whereby naive T cells
a
direct their own phenotype commitment.
gp190 52
IL-6 PRODUCED BY MACROPHAGES, T CELLS, AND B CELLS DURING
RESPONSES TO Ag DRIVES B CELL EXPANSION AND Ig CLASS SWITCHING

IL-6 IS REQUIRED FOR B CELL DIFFERENTIATION INTO PLASMA CELLS.

AUTOCRINE OR PARACRINE
IL-6 DRIVES Ig CLASS IgG2A
IgM IgG1
SWITCHING

AUTOCRINE OR PARACRINE
IL-4 OR IL-13 DRIVES IgG1 IgE
Ig CLASS SWITCHING
TO IgE

53
 TNF FAMILY
The TNF family includes cell surface proteins, such as CD40.
TNFa is the principle cytokine that mediates acute
inflammation. In excessive amounts it also is the principal
cause of systemic complications such as the shock cascade.

Major functions of TNFa:

acts on endothelial cells to stimulate inflammation and coagulation
stimulates endothelial cells to produce selectins and ligands for leukocyte
integrins during diapedesis
stimulates endothelial cells and macrophages to produce chemokines
stimulates macrophages to secrete IL-1
2 types of receptors activates neutrophils
induces acute phase response by liver
cachectic factor (muscle proteolysis and fat catabolism)
cytotoxic for some tumor cells
interacts with hypothalamus to induce fever and sleep
stimulates the synthesis of collagen in wound healing and tissue repair
activates macrophages and dendritic cells
is required by thymocytes for normal T cell differentiation

TNFa is produced by monocytes, macrophages, dendritic cells, TH1 cells,

and others. TNFa is secreted and also active in membrane-bound form.
54
TNF Receptors
2 types of TNF receptors: one has a Death Domain & is responsible for cytotoxicity.
They are type II membrane proteins.
The other signals through NF-B & is responsible for the activation/differentiation properties.
Ligand/receptor complexes trimerize to transduce either signal.

Lymphotoxin-a (LTa) = TNFb

LTa plays a role in the recruitment and activation of neutrophils and in lymphoid
organogenesis. Produced by T cells, including Tc; plays a role in Tc function. Also
produced by NK cells and B cells.

Lymphotoxin-b (LTb)

Is not secreted, although a specific receptor exists. Active as membrane-bound

cytokine. Aside from cytotoxic properties, it is essential for the formation of
germinal centers.

55
 IL-10 FAMILY
Recently a family of related cytokines has
emerged, comprising a series of herpesviral and
poxviral members and several cellular sequence
homologs, including IL-10, IL-19, IL-20, IL-22, IL-
24 and IL-26. Although the predicted structure
of these molecules is conserved, certain
receptor-binding residues are variable and define
the interaction with specific heterodimers of
different type-2 cytokine receptors.

IL-10 was discovered initially as an inhibitory factor for the production of TH1 cytokines.
Subsequently, pleiotropic inhibitory and stimulatory effects on various types of blood cells were
described for IL-10, including its role as a survival and differentiation factor for B cells. IL-10,
which is produced by activated monocytes and T cells, as well as other cell types, such as
keratinocytes, appears to be a crucial factor for at least some forms of peripheral tolerance and
a major suppressor of the immune response and inflammation. The inhibitory function of IL-10
is mediated by the induction of regulatory T cells (CD4+ CD25+).

IL-10 is also a survival and differentiation factor for B cells.

56
IL-22 mediates acute-phase response signals in hepatocytes and IL-20 induces the hyper-
proliferation of keratinocytes; proposed as a pathogenic mechanism of psoriasis.

Many immunosuppressive and immunostimulatory functions described for IL-10;

biological activities of the others are distinct from those of IL-10, at least as far as is known
for IL-20, IL-22/IL-TIF and IL-24/MDA-7. Moreover, the site of expression is distinct for each.
IL-10 is produced by TH2 cells, macrophages, keratinocytes.

The expression patterns are more restricted for the others.

IL-26/AK155 - monocytes and various types of T cells
IL-22/IL-TIF - CD4+ T cells
IL-24 - melanocytes, LPS-stimulated monos, TH2 cells after stimulation with anti-CD3 + IL-4
IL-19 - LPS-treated monos, B cells.
IL-20 - unclear-- low levels only in some tissue samples from skin and trachea .

57
 IL-12 FAMILY
IL-12
Heterodimer: p35 and p40 Receptor: b1 and b2 subunits
Acts in a contrasting manner to IL-4; promotes TH1 differentiation; induces IF-g
production

Primary mediator of early innate immune responses to intracellular microbes

Inducer of cell-mediated immunity
Stimulates synthesis of IF-g by T cells & NK cells
Increases the killing activity of CTLs & NK cells
Stimulates differentiation of naive CD4+ T cells into IF-gproducing TH1 cells.
Produced mainly by macrophages and dendritic cells.

IL-23
Heterodimer: p19 and p40 (same p40 as IL-12) Receptor: b1 and IL-23-specific subunits

Produced by activated DCs

Induces synthesis of pro-inflammatory cytokines (eg, IL-17) by CD4+ TH1 cells
Does NOT induce IF-g production
Implicated in many autoimmune disorders
IL-23-specific subunit of receptor homologous to IL-12 b2 subunit
May play a key role in innate immunity to microbial pathogens
58
Collison & Vignali.
IL-35: Odd one out or part of the family?
Immunol. Rev. 226(1): 248-262, 2009.

IL-27 is related to both IL-12

and IL-6 cytokine families.

59
IL-23 required for terminal differentiation of TH17 cells

Nature Immunology 10, 236 - 238 (2009)

Don't leave home without it: the IL-23 visa to TH17 cells
Yeonseok Chung & Chen Dong
Interleukin 23 is tightly associated with TH17 cellmediated inflammation and
autoimmunity. A new study of mice deficient in its receptor shows that interleukin 23
is required for the terminal differentiation of TH17 cells in vivo.

Nature Immunology 10, 314 - 324 (2009)

The interleukin 23 receptor is essential for the terminal differentiation of
interleukin 17producing effector T helper cells in vivo
Mandy J McGeachy
Whether interleukin 23 (IL-23) affects the differentiation of or simply the maintenance of TH17
cells is unclear. The data in this paper by McGeachy and colleagues show that IL-23 is
required for the full differentiation and proliferation of effector TH17 cells in vivo.

60
IL-35

Heterodimer: p35 and EBI3* Receptor: UNKNOWN

Inhibits T cell differentiation; expressed by Tregs

Expands Tregs; inhibits TH17cells; has therapeutic effects in collagen arthritis
model. Tregs from IL-35 KO mice fail to ameliorate inflammatory bowel disease in
mouse model.

*EBI3 = Epstein-Barr virus Induced gene 3

61
IL-14

Induces B-cell proliferation, inhibits Ig secretion, and selectively expands memory B cells
Produced by T cells

Has homology with complement factor Bb

IL-16
Soluble ligand to the CD4 molecule (its receptor)
Chemotactic properties for CD4+ cells
Growth factor for CD4+ T cells, upregulating MHC class II and the IL-2 receptor CD25.
Possible sources include CD8+ T cells, eosinophils, macrophages, mast cells

62
 IL-17 FAMILY
IL-17 (IL-17A) - secreted exclusively by a subset of T cells -- TH17 cells

Human IL-17A gene product - 150 amino acids with a MW of 15 kDa, secreted as disulfide-
linked homodimer of 3035 kDa

5 related cytokines share 2050% homology to IL-17

IL-17 - designated IL-17A to indicate that it is the founding member of the IL-17 cytokine family
IL-17F - produced primarily by activated memory T cells
IL-17B, IL-17C, IL-17D, and IL-17E - secreted by a wider assortment of cells
IL-17R - transmembrane proteins of ~130 kDa; expressed on nearly all cells (IL-17RA-D)
IL-17A induced by IL-23; plays proinflammatory role in joint destruction of RA

Signaling by IL-17 via IL17 receptor plays an important role in host defense against pathogens
(J. Exp. Med. Published online Feb. 9, 2009 doi:10.1084/jem.20081463)

IL-25 (aka IL-17E)

Involved in development of TH2 responses

Involved in expulsion of helminthic parasites

Amplifies/exacerbates allergic responses

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Taken from Gaffen. An Overview of IL-17 function & signaling. Cytokine. 43(3): 402-407, 2008.

IL-17 family ligands Produced by Binding receptor(s)

IL-17 (IL-17A, CTLA-8) T cells (TH17) IL-17RA, IL-17RC (IL-17RL)
IL-17A/F heterodimer T cells IL-17RA, IL-17RC
IL-17B Numerous tissues IL-17RB?
IL-17C Prostate, fetal kidney
IL-17D Numerous tissues
IL-17E/IL-25 Numerous tissues IL-17RB (IL-25R)
IL-17F T cells IL-17RA, IL-17RC
vIL-17 (ORF13) H. Saimiri IL-17RA, IL-17RC?
Unknown IL-17RD
Unknown IL-17RE

It is currently believed that IL-17 regulates innate immunity. Activates PMNs

and expression of anti-microbial genes.

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Gaffen. An Overview of IL-17 function & signaling.
Cytokine. 43(3): 402-407, 2008.

65
 IL-27 & IL-30

IL-27 - related to both IL-6 and IL-12 families (which are related to each
other by receptor homology)

IL-30 is p28 subunit of IL-27

IL-27 opposes actions of IL-23, thus has potent anti-inflammatory activity

Predict that IL-27 inhibits TH17 differentiation & maintenance.

Receptor is believed to consist of 2 peptides, gp130 and WSX-1 (aka IL-27R)

(For this reason, one might include these 2 in the IL-6 family)

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 IL-28 & IL-29

IL-28 and IL-29 are a recently discovered family of novel class II cytokines
distantly related to IF-a and IL-10.

IL-29 aka IF-1 & discovered by UMDNJ faculty member Serge Kotenko
IL-28 aka IF-3

Both have antiviral activity against encephalomyocarditis and other viruses.

Receptor is composed of two chains, IL-28R and IL-10Rb

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 IL-35 AND COUNTING

IL-32 is the name given to the NK4 transcript first reported in IL-2 activated T
lymphocytes and natural killer cells 13 years ago without known function. The
novel cytokine has six isoforms.

Dinarello CA & Kim SH. IL-32, a novel cytokine with a possible role in disease.
Ann Rheum Dis. 2006. Nov;65 Suppl 3:iii61-iii64.

no homology with other cytokines

induced by IL-2 and IL-18
induces IF-a, IL-8, MIP-1a
induces IL-1b and IL-6 through a caspase-dependent mechanism

IL-33: see IL-1

IL-34: next slide

IL-35: see IL-12

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IL-34

IL-34 is known also as FPT025.

The protein does not show sequence homology with known cytokines or growth factors.
It is expressed in spleen, skin, brain, and other tissues. IL-34 human primary monocyte
proliferation and/or survival. Human IL-34 specifically binds to human primary monocytes and
activates ERK1/2 phosphorylation in a human monocytic cell line, THP-1. IL-34 regulates
myeloid lineage differentiation, proliferation, and survival; promotes CFU-GM and CFU-M
colony formation from human bone marrow cells in a colony formation assay; stimulates
expression of myeloid cell surface markers. IL-34 appears to act via the receptor for
M-CSF.

Lin H et al Discovery of a cytokine and its receptor by functional screening of the extracellular
proteome. Science 320(5877): 807-811 (2008).

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CYTOKINE CASCADE LINKS INFLAMMATORY AND IMMMUNE
RESPONSES

IMMUNE DANGER SIGNAL INFLAMMATORY

MICROORGANISM

NEUTROPHIL
MONOCYTE, MACROPHAGE

IL-1, TNF
ANTI-MICROBIAL MECHANISMS
IL-18 (ROI, RNI, etc.)
B CELLS
DENDRITIC CELLS

IL-12 IL-6
T CELLS IL-4 Ab:
Enhanced Phagocytosis
Activation of Complement
ADCC
IL-2
TH1, TC

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CYTOKINES DETERMINE NATURE OF ADAPTIVE IMMUNE RESPONSE

IL-12, IF-g
TH1: IL-2, TNF, IF-g

TH0 TH1
APC

TH2

IL-4
IL-4

eosinophil TH2 TH2: IL-4, IL-5, IL-6, IL-13

basophil

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72