CYTOKINES

Cytokines
• A collection of polypeptides used for
communications between cells
• Low molecular weight
• secreted by cells of the immune system
• Affect the behaviour of other cells
• Role similar to hormones-Signalling molecules
• Key players in innate and acquired immunity
 Which cells release cytokines ?
Cells of the immune system:
• Neutrophils – when they
encounter a pathogen

• Macrophages – when they

encounter a pathogen

• NK cells – on encountering a
microbe infected cell /tumour cell

• Lymphocytes – when they are

activated
 Cytokine Nomenclature
⮚ Originally were called lymphokines because they were
initially thought to be produced only by lymphocytes.
Then monokine because they were secreted by
monocytes and macrophages. Then interleukin because
they are produced by some leukocytes and affect other
leukocytes.
⮚ The term “cytokine” is now used more widely and covers
all of the above. Chemokines, are also considered
cytokines.
1. Interleukins (1-18)
2. Interferons (α,β,γ)
3. Tumour necrosis factor (TNF)
4. Chemokines
• Subpopulation of cytokines
• Move immune cells from place to place
 Cytokine -mediated effects

• Cell growth

• Cell differentiation

• Cell death

• Induce non-responsiveness to other

cytokines/cells

• Induce responsiveness to other cytokines/cells

• Induce secretion of other cytokines

How do cytokines tell cells what to do?

• Produced by cells as part of

normal cellular activity and/or
the result of environmental
trigger

• Bind to receptors on cells

• Trigger signal transduction

pathways

• Signal results in altered pattern

of gene expression

• Initiate synthesis of new proteins

 Cytokines can act in three different
manners

• Autocrine
– Cytokine binds to
receptor on cell that
secreted it
• Paracrine
– Cytokine binds to
receptors on near by cells
• Endocrine
– Cytokine binds cells in
distant parts of the body
 Cytokine Actions
1. Pleiotropy
– Act on more than one cell type (INFα/β)
2. Redundancy
– More than one cytokine can do the same thing
(IFNα/β and IFNγ)
3. Synergy
– Two or more cytokines cooperate to produce an
effect that is different or greater than the combined
effect of the two cytokines when functioning
separately (IL-12 and IL-8)
4. Antagonism
– Two or more cytokines work against each other (IL-4
and IL-12)
1. Cytokines are pleiotropic … one cytokine can have
different effects on different cells.
2. Cytokines can be redundant … different cytokines can
have the same effects.
3. Cytokines can synergize with each other.
4. Cytokines can antagonize each other.
5. Cascade effect, cytokines can stimulate the
production of other cytokines.
6. Cytokines can influence the expression of
cytokine receptors.
 7. Cytokines play key roles in regulating
hematopoiesis, innate immunity and acquired
immunity.
There are many cytokines, including...
IL-1 IL-2 IL-3 IL-4
IL-5 IL-6 IL-7 IL-8
IL-9 IL-10 IL-11 IL-12
IL-13 IL-15 IL-16 IL-17
IL-18 IL-19 IL-20 IL-21
IL-22 IL-23
IFN-a IFN-b IFN-g
TNF-a TNF-b
TGF-b1
M-CSF G-CSF GM-CSF
 Cytokine Families
1. IL-1 family
2. Hematopoietin family
3. Interferon family
4. TNF family
5. IL-17 family
6. Chemokines family
 IL-1 family
• IL-1 first noninterferon cytokine to be
identified.
• Important proinflammatory mediators.
• E.g. IL-1, IL-18, IL-33
 Hematopoietin family
– α-helical structure
prevalence
– Little or no β-
sheet
– Ex. IL-2 and IL-4
– Amino acid
sequences vary
considerably
 Interferon family
• Antiviral response
• Immunomodulators
• E.g. IFN-α/β/γ, IL-10
 Interferons (IFN)
• Cytokines produced by virus infected monocytes and
lymphocytes
• “Interfere” with virus replication
• Warn the neighbouring cells that a virus is around...
1. Viral replication stimulates the infected host cell to produce
interferon.
2. Interferon induces uninfected cells to
– produce antiviral proteins that prevent translation of viral mRNA
– degrade viral nucleic acid
3. Viral replication is blocked in uninfected cells
• If we did not have IFNs – most of us may die of influenza virus
infection
How does IFN warn the neighbouring
cells ?

22
The infected cells release IFN

antiviral state

antiviral state antiviral state

antiviral state
23
Virus infects the neighbouring cells

antiviral state

antiviral state antiviral state

antiviral state
24
Prewarned cells are able to quickly
inhibit the virus

antiviral state

antiviral state antiviral state

antiviral state
25
 How do Interferons inhibit viruses ?

Inactive host protein

Induction Virus ds-RNA
Activation

Host protein Active host protein

Cascade of events

Inhibition of
host protein
synthesis
Virus cannot replicate
 TNF family
• Soluble or membrane bound
• Immune system development, effector
functions and homeostasis
• E.g. TNF-α/β, CD40L, Fas (CD95)
 Tumour necrosis factor (TNF)
TNF

Killing of cancer Fever

Inflammation
 IL-17 family
• Recently discovered
• Proinflammatory
• Promote neutrophil accumulation and
activation
• E.g. IL-17 (A/B/C/D/F)
 Chemokines family
• Chemoattractants: Recruit to sites of infection
• MIP-1α (NK and T cells)
• MIG, RANTES (CD4+T cells)
• IL-8 (neutrophils)
• Eotaxin (eosinophils)
Best way to learn about cytokines…. Is
by their action !!!
 Early mediators

• Interferons α/β
– Induced by dsRNA, etc.
– IFNs can induce more of themselves
– Directly interferes with viral replication
– Activation of T and NK cells
 Early mediators
• IL-12, IL-15, 1l-18, IFN-γ (from NK cells), IL-10
• Proinflammatory mediators
• Produced by cell associated with innate immunity
(macrophages, NK, etc.)
• Mediate direct effects
• Promote inflammation
• Shape downstream responses
 Late mediators

• IL-2, IL-4, IL-5, IFN-γ, TNF, IL-6, IL-10

• Produced by cells of the adaptive immune
response (T and B cells)
• Direct effects
• More immunoregulatory functions
 Down regulators

• IL-10, IL-11, TGF-β

• Inhibit proliferation, cytokine
production
• Produced by both innate and adaptive
cells
 Maintenance cytokines

• GM-CSF, IL-3, IL-7, IL-9, etc.

• Induce cell differentiation, cell growth
 The Jak/Stat Signaling
Pathway
• A number of cytokine receptors signal via the
JAK/STAT pathway.
• These include the receptors for IL-2, IL-3, IL-4, IL-6,
IL-10, IL-12 and IFN-g.
• Cytokine receptor subunits are associated with JAK
kinases.

– JAK = Janus Kinase - OR – Just Another Kinase

– STAT = Signal Transducers and Activators of Transcription
STATs (signal
transducers and
activators of
transcription)

Janus
kinases
(JAKs)
 The Jak/Stat Signaling
Pathway
1.- Binding of cytokine causes
dimerization of receptors and
activation of JAK kinases.
2.- Activated JAK kinases
phosphorylate receptor sites and
create docking sites for STAT
molecules.
3. After binding to the receptor (a
chain), STATs are
phosphorylated.
4. They then dissociate from the
receptor, dimerize and
translocate to the nucleus, where
they mediate transcription of
target genes.
 Cytokine cross-
regulation
• Helper T cells can be divided into two main types -
TH1 and TH2 - with distinct patterns of cytokine
secretion.

• In a a given immune response, either T H1 or TH2

response dominates

• Cytokines of one response tend to down-regulate the

other type of response

• Example: TH1 cells secrete IFN-g, which inhibits

proliferation of TH2 subset
TH1/TH2 differentiation is influenced by the levels of key
cytokines.
- IL-4 promotes TH2 differentiation.
- IFN-g and IL-12 promotes TH1 differentiation.
Cytokine secretion and biological
activities of TH1 and TH2 Subsets
• TH1 cells produce cytokines (IFN-g and IL-2) that promote
immune responses against intracellular pathogens (DTH,
cytotoxic T cell responses, opsonizing Abs).

• TH2 cells produce cytokines (IL-4, IL-5, IL-6, IL-13) that

promote immune responses against extracellular pathogens
(antibody responses, eosinophilic responses, allergic
reactions).

• Some cytokines are produced by both TH1 and TH2 cells.

These cytokines - GM-CSF and IL-3 - act on the bone marrow
to increase production of leukocytes - so they are needed no
matter what type of pathogen is present.
 Cytokine secretion and
biological activities of TH1 and
TH2 Subsets

Type Type
Cell-mediated 2 Humoral
1
Immune response T cell response
(Intracellular (extracellular
pathogens) IL-2 IL-4 pathogens)
IFN-g IL-5
TNF
 Role of TH1/TH2 balance in
determining disease outcomes
• Balance of two subset determines response to
disease
• Relative predominance of TH1 vs TH2 helper T
cells can influence the course of infectious disease
(Mycobacterium leprae)
• Leprosy
– Tuberculoid (TH1, CMI response, patient lives)
– Lepromatous (TH2, humoral response, patient
dies)
 Course of Leprosy

Tuberculoid - CMI (granulomas) Lepromatous - HI (dissemination)

No RIP RIP
 Cytokine-related
diseases
• Bacterial septic shock
– Blood pressure drops, clots form, hypoglycemia ensues, patient dies
– LPS triggers results in TNF release
– TNF induces IL-1 which induces IL-6 and IL-8

• Bacterial toxic shock and related diseases

– Superantigens trigger large numbers of T cells which release massive
amounts of cytokines (Super antigens are bacterial toxins that bridge
CD4 T cell receptors and the MHC class II molecules on APC’s,
bypassing the need for antigen)

• Lymphoid and myeloid cancers

– Some cancer cells secrete cytokines

• Chagas’ disease
– Trypanosoma cruzi infection results in sever immune suppression
– Depression of IL-2 receptor production
Therapeutic uses of cytokines
• Modulation of T H activation
• Interfere with receptor function
• Interfere with cytokine
– Make it unable to bind to receptor
– Make it unable to act
• Examples
– Anti-IL-2R
– Interleukin analogs which bind receptor, but do
not trigger activation (ties up receptor)
– Toxins conjugated to cytokines which kill
activated T-cells
– Administration of cytokines to enhance
immunity (side effects/ short half lives)