CYTOKINES

“Cytokine" is a word that comes from cyto- a combining form meaning "cell" - and -kinin - a
combining form used in naming hormones, especially peptide hormones (e.g., bradykinin).

Cytokines are a group of low molecular weight regulatory proteins secreted by white blood
cells as well as a variety of other cells in the body in response to inducing stimuli. 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.

Properties of Cytokines

1.Cytokines are small secreted proteins which mediate and regulate immunity, inflammation
and haematopoesis.

2. They must be produced de novo in response to an immune stimulus.

3. They generally (although not always) act over short distances and short time spans and at
very low concentration.

4. They act by binding to specific membrane receptors, which then signal the cell via second
messengers (often tyrosine kinases) to alter its behavior (gene expression).

5.Responses to cytokines include increasing or decreasing expression of membrane proteins

(including cytokine receptors), proliferation, and secretion of effector molecules.

Cytokines are a unique family of growth factors. Secreted primarily from leukocytes, cytokines
stimulate both, the humoral and cellular immune responses, as well as the activation of
phagocytic cells. Growth factors are proteins that bind to receptors on the cell surface, with the
primary result of activating cellular proliferation and/or differentiation. Many growth factors
are quite versatile, stimulating cellular division in numerous different cell types; while others
are specific to a particular cell-type.

The following table lists some of the more commonly known factors and their principal
activities.
 Principal
Factor Primary Activity
Source

platelets,
promotes proliferation of connective
PDGF endothelial cells,
tissue, glial and smooth muscle cells
placenta

submaxillary
promotes proliferation of
EGF gland, Brunners
mesenchymal, glial and epithelial cells
gland

common in may be important for normal wound

TGF-a
transformed cells healing

wide range of
promotes proliferation of many cells;
cells; protein is
FGF inhibits some stem cells; induces
associated with the
mesoderm to form in early embryos
ECM

promotes neurite outgrowth and neural

NGF
cell survival

promotes proliferation and

Erythropoietin kidney
differentiation of erythrocytes

anti-inflammatory (suppresses
activated TH1 cells
cytokine production and class II MHC
(T-helper) and
TGF-b expression), promotes wound healing,
natural killer (NK)
inhibits macrophage and lymphocyte
cells
proliferation

promotes proliferation of many cell

IGF-I primarily liver
types

promotes proliferation of many cell

IGF-II variety of cells
types primarily of fetal origin

Cytokine is a general name; other names include lymphokine (cytokines made by

lymphocytes), monokine (cytokines made by monocytes), chemokine (cytokines with
chemotactic activities), and interleukin (cytokines made by one leukocyte and acting on other
leukocytes). Cytokines may act on the cells that secrete them (autocrine action), on nearby
cells (paracrine action), or in some instances on distant cells (endocrine action).It is common
for different cell types to secrete the same cytokine or for a single cytokine to act on several
different cell types. Cytokines are redundant in their activity, meaning similar functions can
be stimulated by different cytokines. Cytokines are often produced in a cascade, as one
cytokine stimulates its target cells to make additional cytokines. Cytokines can also act
synergistically (two or more cytokines acting together) or antagonistically (cytokines causing
opposing activities).

Their short half life, low plasma concentrations, pleiotropy, and redundancy all complicated
the isolation and characterization of cytokines.

Selected Immune Cytokines and Their Activities*

Cytokine Producing Cell Target Cell Function**
growth and differentiation of
GM-CSF Th cells progenitor cells
monocytes and DC
Th cells co-stimulation
monocytes B cells maturation and proliferation
IL-1a macrophages
IL-1b B cells NK cells activation
DC inflammation, acute phase
various
response, fever
activated T and B cells, growth, proliferation,
IL-2 Th1 cells
NK cells activation
Th cells stem cells growth and differentiation
IL-3
NK cells mast cells growth and histamine release
proliferation and differentiation
activated B cells of
IL-4 Th2 cells IgG1 and IgE synthesis
macrophages MHC Class II
T cells proliferation
proliferation and differentiation
IL-5 Th2 cells activated B cells
IgA synthesis
activated B cells differentiation into plasma cells
monocytes
macrophages plasma cells antibody secretion
IL-6
Th2 cells stem cells differentiation
stromal cells
various acute phase response
marrow stroma differentiation into progenitor B
IL-7 stem cells
thymus stroma and T cells
macrophages
IL-8 neutrophils chemotaxis
endothelial cells
 macrophages cytokine production
IL-10 Th2 cells
B cells activation
differentiation into CTL
macrophages activated Tc cells
IL-12 (with IL-2)
B cells
NK cells activation
viral replication
IFN-a leukocytes various
MHC I expression
viral replication
IFN-b fibroblasts various
MHC I expression
various Viral replication
macrophages MHC expression
Th1 cells,
IFN-y activated B cells Ig class switch to IgG2a
Tc cells, NK cells
Th2 cells proliferation
macrophages pathogen elimination
MIP-1a macrophages monocytes, T cells chemotaxis
MIP-1b lymphocytes monocytes, T cells chemotaxis
monocytes,
chemotaxis
macrophages
TGF-b T cells, monocytes activated macrophages IL-1 synthesis
activated B cells IgA synthesis
various proliferation
macrophages, mast macrophages CAM and cytokine expression
TNF-a
cells, NK cells tumor cells cell death
phagocytes phagocytosis, NO production
TNF-b Th1 and Tc cells
tumor cells cell death

* CTL: cytotoxic T lymphocytes; DC: dendritic cells; GM-CSF: Granulocyte-Monocyte

Colony Stimulating Factor; IL: interleukin; IFN: Interferon; TGF: Tumor Growth Factor;
TNF: Tumor Necrosis Factor.
** Italicized activities are inhibited.

Classification Of Cytokines

The most common cytokines classified on the basis of their general properties
1. Cytokines that mediate and regulate innate immunity
1.1.1. Type I interferons
1.1.2. Tumor necrosis factor-a
1.1.3. Interleukins 1, 6, 10, 12, and 15
1.1.4. Chemokines
2. Cytokines that mediate and regulate specific immunity
2.1.1. Interleukins 2, 4, 5, 13, 16, and 17
2.1.2. Interferon-g
2.1.3. TGF-ß
2.1.4. Lymphotoxin
3. Cytokines that stimulate hematopoiesis
3.1.1. c-kit ligand
3.1.2. Interleukins 3, 7, 9, 11
3.1.3. Colony stimulating factors (CSF)

RECEPTORS TO WHICH CYTOKINES BIND

1. Immunoglobulin superfamily
2. Cytokine receptor family - Class I
3. Cytokine receptor family - Class II
4. TNF receptor family
5. Seven transmembrane helix family

The immunoglobulin superfamily is characterized by one or more Ig domains which are

regions of 70 to 110 amino acid residues homologous to either Ig V or C domains. Examples
include receptors for IL-1 and M-CSF.

CLASS I CYTOKINE RECEPTOR FAMILY

(HEMATOPOIETIN RECEPTOR FAMILY)

Most of the cytokine-binding receptors that function in the immune and hematopoietic systems
belong to this receptor family. In addition, this family includes receptors for growth hormone
and prolactin. The receptors consist of 2 polypeptide chains: a cytokine-specific subunit and a
signal-transducing subunit which is usually not specific for the cytokine. In a few cases these
receptors are trimers. The signal transducing subunit is required for high affinity binding of
the cytokine.

Class I cytokine receptors have been further divided into sub-families with all the receptors in
one subfamily having an identical signal transducing subunit.

GM-CSF SUBFAMILY
This subfamily includes the receptors for IL-3, IL-5, and GM-CSF. The unique low affinity,
cytokine-specific receptor is the a subunit. All three low affinity a subunits can associate
noncovalently with a common signal transducing ß subunit. The resulting dimeric receptor
exhibits increased affinity for the cytokine but also transduces a signal across the membrane
following cytokine binding. Interestingly, IL-3, IL-5, and GM-CSF exhibit considerable
redundancy in their activities.

IL-6 SUBFAMILY
This subfamily includes the receptors for IL-6, IL-11, and IL-12. In this case a common signal
transducing subunit (gp130) associates with one or two different cytokine-specific subunits.
As expected, the cytokines that bind to receptors in this subfamily display overlapping
biological activities.

IL-2 SUBFAMILY

This subfamily includes the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15.

The IL-2 and IL-15 receptors are trimers, consisting of a cytokine-specific a chain and two
chains - ß and g - responsible for signal transduction. The IL-2 receptor g chain is the signal
transducing subunit in those members of this subfamily which are dimers.

CLASS II CYTOKINE RECEPTOR FAMILY (INTERFERON RECEPTOR

FAMILY)
The ligands for these receptors are the three interferons a, ß, and g. Signal transduction through
most, if not all, class I and class II cytokine receptors begins with cytokine-induced
dimerization of receptor subunits. This dimerization puts the cytosolic parts of the receptor
closer together, leading to engagement of the intracellular signaling machinery. It had been
thought that the molecular machinery of cytokine signaling was located only in the cytoplasm.
Recently evidence has been accumulated - and it is still somewhat controversial - that
cytokines may also be targeted to the cell nucleus.

TUMOUR NECROSIS FACTOR SUPERFMILY

The TNF receptor family of transmembrane glycoproteins includes the 55 kDa TNF receptor
(TNF-RI) and the 75 kDa TNF receptor (TNF-RII), as well as CD40 and Fas.

Tumor Necrosis Factor-a (TNF-a)

TNF-a (also called cachectin), like IL-1 is a major immune response-- modifying cytokine
produced primarily by activated macrophages. Like IL-1, TNF-a induces the expression of
other autocrine growth factors, increases cellular responsiveness to growth factors and induces
signaling pathways that lead to proliferation. TNF-a acts synergistically with EGF and PDGF
on some cell types. Like other growth factors, TNF-a induces expression of a number of
nuclear proto-oncogenes as well as of several interleukins.

Tumor Necrosis Factor-b (TNF-b)

TNF-b (also called lymphotoxin) is characterized by its ability to kill a number of different cell
types, as well as the ability to induce terminal differentiation in others. One significant non-
proliferative response to TNF-b is an inhibition of lipoprotein lipase present on the surface of
vascular endothelial cells. The predominant site of TNF-b synthesis is T-lymphocytes, in
particular the special class of T-cells called cytotoxic T-lymphocytes (CTL cells). The
induction of TNF-b expression results from elevations in IL-2 as well as the interaction of
antigen with T-cell receptors.

WHAT IS A CHEMOKINE?

Chemokines are a family of structurally related glycoproteins with potent leukocyte activation
and/or chemotactic activity. They are 70 to 90 amino acids in length and approximately 8 to
10 kDa in molecular weight. Most of them fit into two subfamilies with four cysteine
residues. These subfamilies are base on whether the two amino terminal cysteine residues are
immediately adjacent or separated by one amino acid. The a chemokines, also known as CXC
chemokines, contain a single amino acid between the first and second cysteine residues; ß, or
CC, chemokines have adjacent cysteine residues. Most CXC chemokines are chemoattractants
for neutrophils whereas CC chemokines generally attract monocytes, lymphocytes, basophils,
and eosinophils. There are also 2 other small sub-groups. The C group has one member
(lymphotactin). It lacks one of the cysteines in the four-cysteine motif, but shares homology at
its carboxyl terminus with the C-C chemokines. The C chemokine seems to be lymphocyte
specific. The fourth subgroup is the C-X3-C subgroup. The C-X3-C chemokine
(fractalkine/neurotactin) has three amino acid residues between the first two cysteine. It is
tethered directly to the cell membrane via a long mucin stalk and induces both adhesion and
migration of leukocytes.
CHEMOKINE RECEPTOR FAMILY

The chemokine receptors are members of a superfamily of seven transmembrane loops and
transduce their signals through heterotrimeric G proteins.

Chemokine receptors are structurally related and can be categorized into specific (bind only
one known ligand - e.g., CXCR1/IL8RA and CXCR4/fusin/LESTR), shared (CXCR2/IL8RB,
CXCR3, CCCR1-CCCR5), promiscuous (bind to many chemokine ligands of either CXC or
CC types - e.g. Duffy blood group antigen), and viral (shared receptors that have been
transduced into viral genomes during evolution - herpes saimiri virus and cytomegalovirus).
The N-terminal portion of chemokine receptors is key to determining ligand binding
specificity.

If engagement of chemokine receptors results in the movement of the cell, a complex series of
signaling circuits are involved. Different pathways lead to activation and proliferation.

Mechanism for Downregulation of Cytokines

Clearly it is necessary that there be mechanisms by which cytokines are downregulated.

Examples include:

(1) Cytokine antagonists such as the IL-1 receptor antagonist (IL-1Ra) which binds to the IL-1
receptor. Antagonists bind to a specific receptor but do not transmit a signal. Production of
IL-1Ra appears to play a role in regulating the intensity of the inflammatory response.

(2) Soluble cytokine receptors can be found in the blood and extracellular fluid. These soluble
receptors result from enzymatic cleavage of the extracellular domain of cell-bound cytokine
receptors. The released soluble fragments can bind cytokine molecules, thereby neutralizing
their activity. The soluble IL-2 receptor (sIL-2R), which is released following chronic T cell
activation is the best characterized. The shed receptor can bind IL-2 and prevent its interaction
with the membrane-bound IL-2R.
(3) Other cytokines, acting through quite separate receptors, could exert opposite effects on
cells.

(4) Cytokines could bind to receptors that do not activate the cell.

Clinical uses of cytokines

Given the large number of cytokines, it is disappointing that relatively few can be used
clinically. Generally, cytokines with more restricted target-cell spectra are more likely to be
useful for treatment. Examples include:

a) Colony stimulating factors (CSFs) - hematological disorders associated with cancer

therapy
b) Erythropoietin (EPO) - anemia associated with kidney disease
c) IFN-ß - multiple sclerosis
d) IFN-g - chronic granulomatous disease (GCD)
e) Growth hormone - growth hormone deficiency
f) IL-2 - kidney cancer, melanoma
g) IL-11 - thrombocytopenia following high dose chemotherapy

P.S: Cytokine antagonists are being considered for use as treatments for diseases
associated with overproduction of cytokines - such as bacterial septic shock (gram
negative bacteria with overproduction of TNF-a or IL-1) or bacterial toxic shock
(bacterial superantigens with overproduction of TNF-a or IL-1).