Introduction to immunology.

LESSON 4: CYTOKINES, CHEMOKINES AND IMMUNE RESPONSES

Today we will get to know:

• Cytokines and chemokines

• The major signaling pathways in the

immune system and what they do

• “Good” and “bad” immune reactions

• A little bit of hematology

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines and chemokines

Cytokines and chemokines are a very wide group of molecules whose role is to stimulate or
suppress the functions and the proliferation of immune cells (cytokines) or to attract
immune cells to districts where they are needed (chemokines).

CYTOKINES CHEMOKINES
What to do?
Where to go?
Should we
proliferate? Should we stay?

Systemic
responses?

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines

Cytokines mediate and regulate all the aspects of innate and adaptive immune responses.
There are at least 180 genes in the human genome which code for cytokines… and, their
nomenclature is a mess!

CYTOKINES

By function (i.e., By the producing cell

Interferons or TNF) (i.e., Interleukins - IL)

“The mediators that exert this “The mediators produced by this

particular effect” particular cell”

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines

All the cytokines share some common features:

• Generally, they are synthesized ex-novo

after gene activation. Only a very few
cytokines are stored in the cytoplasm and
ready to be released.
• They are small. Generally, their weight is < 30
kD.
• They are pleiotropic. A single cytokine can
act on different target cells and exert
different actions.
• They are redundant. Many different
cytokines can exert the same function.
• They are integrated. Different cytokines can
synergize or antagonize each other.
• They are always released. Cytokines are
extracellular mediators.

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines

They are always released. Cytokines are extracellular mediators.

AUTOCRINE PARACRINE ENDOCRINE

Upon release, the cytokine Upon release, the cytokine Only when their quantity is
acts on the same cells acts on surrounding cells, very high, some cytokines
which produced it, influencing their behavior. can enter the blood flow
influencing its behavior. and influence distant
organs.

Physiological Physio-pathological

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Monokines, lymphokines and interleukins

All immune cells are able to produce cytokines. Many non-immune cells are able to produce
them, too. Nevertheless, macrophages and Th cells are the major producers.

• Monokines are the cytokines

produced by macrophages.
• Lymphokines are the cytokines
produced by lymphocytes.
• Interleukins are the cytokines which
act as mediators in between the
leukocytes.

Jean Kelly.

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines and specificity

Cytokines are not antigen-specific. Nevertheless, they mediate all the communications
within the immune system. So, how do we keep the communication specific?

Antigen recognition is specific

PAMPs and DAMPs by PRRs,
Peptides by MHC/TCR
All other by BCR

Cytokine receptors are specific

And specifically expressed by
immune cells

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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokines and specificity

Cytokines induce themselves in a cascade mode. Often, their cascade amplification also
involves different cell types.
Cytokines are needed to connect innate and adaptive
immune responses. They regulate the timing of the
response, integrate the two systems, enable reciprocal
empowering and control and, in the end, enable the
shut-down.
IFN-g
Ensuring a specific communication is essential! Spatial
confinement is the most-effective way to reduce off-
target risk.

IFN-g

Cytokines Cytokines Cytokines

have a short release is release is
half-life very local polarized
IFN-g, TNF-a,
IL-2, etc. 8
 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokine families

Th17 cells Th2 cells M1 M2

Macrophages Macrophages
IL-17 IL-4
IL-1 IL-12
IL-21 IL-5
IL-6 IL-6
IL-22 IL-6
IL-12 IL-8
IL-23 IL-13
IL-15 IL-10
IL-10
Th1 cells IL-23 TGF-b
TNF-a Treg cells
TNF-a
IFN-g TGF-b
IL-10 Dendritic cells
IL-2
IL-10 IL-35 IL-1 IL-12
IL-6 IL-23
IL-10 TNF-a
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 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokine receptors

Type-I and –II receptors signal

via Jak/STAT

TNF receptors signal via

multiple adaptors, including
TRAF

IL-1 receptors signal via

Jak/STAT and NF-kB

Abbas et al. 10
 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokine receptors Type-I and –II signaling pathways

Type-I and –II receptors signal via

Jak/STAT. Once STAT is active and
translocated to the nucleus, many
different responsive genes are
activated. As a result we will see:
• Clonal expansion of T- and B-cells,
proliferation of NK and monocytes
• T-cell polarization
• Granulocyte and macrophage
activation
• Isotype switch
• Antiviral defenses activation

TYPE-I AND –II MAINLY MEDIATE

INFLAMMATORY AND ANTI-VIRAL
RESPONSES.

Type-II, engaged by IL-10, will also

shut-down those responses.

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Abbas et al.
 Introduction to immunology. Lesson 4

Soluble mediators of immunity – Cytokine receptors and NF-kB signaling pathways

Many receptors (including TNF-R, TLRs,

TCR etc.) signal via NF-kB. Once Nf-kB
is released from IkBa inhibition, it
translocates to the nucleus, where
many different responsive genes are
activated. As a result we will see:
• Cytokine production
• Myeloid and lymphoid cells
activation
• Pro-inflammatory activation of
immune cells
• Antiviral defenses activation

NF-kB is engaged by TNF-R, IL-1R and

also by Type-I and II receptors. Its
actions are very pleiotropic.

NF-kB can also mediate response shut-

down.
Abbas et al. 12
 Introduction to immunology. Lesson 4

Soluble mediators of immunity – networks and integrations

Quite an intricate network of signals and receptors…

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 Introduction to immunology. Lesson 4

The immune system in action: extracellular microbes

In the case of extracellular

microbes, humoral immunity (Ig
and complement) will provide
recognition. This will generally
facilitate killing of the pathogen,
which is largely due to the
phagocytes.

Helper T-cells will need time to

recognize the antigen and get
activated, and in the end they will
produce cytokines which will
boost the activity of phagocytes
and B-cells.

BUT WHAT IF A MICROBE CAN

REPLICATE INSIDE THE CELLS?

Abbas et al.
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 Introduction to immunology. Lesson 4

The immune system in action: intracellular microbes

In the case of intracellular

Number of living microbes

microbes, innate immunity will

provide first-line defense. This will
generally reduce microbial spread,
allowing the time needed for the
activation of the innate immunity,
which will kill infected cells with
CTLs and opsonize/neutralize
microbes with antibodies.

Innate immunity will also kill

intracellular microbes, provided
that innate immunity cells have
been infected.

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Abbas et al.
 Introduction to immunology. Lesson 4

The immune system in action: inflammation and fever

Inflammation is a local
event (referred to as
acute inflammation), in
which leukocytes and
plasmatic proteins
(largely belonging to the
humoral immunity) are
recruited in the tissues
(thanks to the
chemokines) to kill
pathogens and prevent
infections.

Cytokines control all the

local reactions.

Abbas et al.

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 Introduction to immunology. Lesson 4

The immune system in action: inflammation and fever

Inflammatory cytokines
Fever is triggered by the are pleiotropic and
hypothalamus-liver axis.
interconnected. Besides
Raising the body temperature
slows down microbial
controlling the immune
replication response, if in sufficient
quantity, they will also
Acute phase proteins include trigger systemic
complement factors, protective effects.
pentraxins/collectins/ficolins
and so on, which aid immune
cell to recognize and kill Eradication of the
pathogens pathogen/stimulus and
suppressive feedback
Stimulation of the mechanisms will stop
hematopoietic system strongly the reaction to prevent
increases the number of damage to the host.
available leukocytes
Abbas et al.
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 Introduction to immunology. Lesson 4

Failures of the immune system: chronic inflammation

During inflammation, many dangerous molecules (oxidant and toxic intermediates, lytic
enzymes, etc.) are released by immune cells. Thus, a low level of tissue damage is always
present. When acute inflammation is resolved, wound healing systems will fix the tissue and
restore its normal functions.

BUT
When the immune system is unable to clear the local pathogen/stimulus, inflammation can
go on indefinitely, damaging host tissues (and potentially making more damages than the
stimulus itself). This is a pathological condition known as chronic inflammation. It can depend
on the ability of the pathogen to actively counteract the immune response, the impossibility
of the immune system to destroy the stimulus (i.e., metals) or genetic defects of the innate
immunity.

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 Introduction to immunology. Lesson 4

Failures of the immune system: chronic inflammation

Being deregulated in time and magnitude, chronic inflammation is always detrimental for the
host (it is pathological). Many major clinical manifestations of important infections are,
actually, just the result of chronic inflammatory processes (destruction of host tissues,
necrosis and fibrosis).

The common feature of chronic

inflammation is the granuloma, which
is a ring-shaped area where immune
cells accumulate and proliferate,
destroying the tissue and paving the
way to scar/fibrous repair tissue
formation. The central area of the
granuloma is generally necrotic, and
tissue in there has a few possibilities
to heal normally. Granulomas can
grow massively, destroying big
portions of tissues.

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 Introduction to immunology. Lesson 4

Failures of the immune system: chronic inflammation

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 Introduction to immunology. Lesson 4

Failures of the immune system: chronic inflammation

Different causes and mechanisms can lead to chronic inflammation

Microbes evade the killing The stimulus cannot be Genetic lesions of the
mechanisms eliminated immune system

Inflammation goes on in an Inflammation goes on Inflammation goes on

attempt to kill the
pathogens, creating
granulomas which
dramatically damage the
host (tuberculosis, leprosy,
syphilis, leishmaniasis, etc.)
creating granulomas which
isolate the stimulus from the
host (foreign body
granuloma)
without being effective in
killing microbes, creating
granulomas which damage
the host (CGD because of
ineffective ROS production
by phagocytes)

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 Introduction to immunology. Lesson 4

Failures of the immune system: septic shock and cytokine storm

Epithelia and lymphoid organs are organized as a series of barriers to prevent pathogens
dissemination through the blood. Nevertheless, sometimes microbes manage to spread
through the blood flow. This is a very dangerous condition which requires massive immune
system activation. Septic shock and cytokine storm are the most common problems
associated with these reactions.

LPS and peptidoglycans activate TLRs, which, through

NF-kB signaling, induce the production of pro-
inflammatory cytokines like IL-1 and TNF-a.

SEPTIC SHOCK:
Altered vascular
permeability and energetic
input will translate into
multiple organs failure,
disseminated intravascular
coagulation, heart failure
and so on. 22
Abbas et al.
 Introduction to immunology. Lesson 4

Failures of the immune system: septic shock and cytokine storm

Cytokine storm (also known as hypercytokinemia) occurs when cytokine cascade goes out of
control. This is generally due to antigens able to super-stimulate the immune system, like
those from avian flu, bacterial meningitis, staphylococcal superantigens, smallpox etc.

Staphylococcal superantigens
bind to CDR4 (all other antigens
bind in the region between
CDR1-3), “skipping” TCR
specificity and activating many T-
cell clones at the time

CYTOKINE STORM:
Polyclonal proliferation of T-cells
induces the production of vast
amount of cytokines, which
super-activate other immune
cells and so on until multiple
organs failure occur.
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 Introduction to immunology. Lesson 4

Hematology lab: complete blood count with differential

Hematopoiesis keep the number of red and white blood cells constant throughout life, but has
to be ready to increase those numbers if the need occurs (i.e., during infections). Complete
blood count (CBC) lists the major cell types of the blood and other parameters. “Differential”
counts will divide the overall leukocytes into their major populations.

3.5-10.5 billion
White blood cell cells/L
count (3,500 to 10,500
cells/ml)

When this number is increased:

LEUKOCYTOSIS (generally due to
inflammation, infections, allergies,
drugs) when transient. Otherwise, a
proliferative pathology of the bone
marrow (MYELOPROLIFERATIVE
DISORDERS AND LEUKEMIA) must be
suspected.
When this number is decreased:
LEUKOPENIA (generally due to drugs
and some medications, autoimmune
problems, myelodysplastic syndromes
and cancer)
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 Introduction to immunology. Lesson 4

Hematology lab: differential counts

Differential WBC counts and common causes of variation
Increase Decrease
Acute infections, acute stress,
Aplastic anemia, influenza,
thyroiditis, leukemia,
Neutrophils 40%-60% rheumatoid arthritis and
chemo-radiotherapy, tumors,
severe bacterial infections
rheumatic fever

Chronic bacterial infections and

Chemo-radiotherapy, tumors,
some viral infections
Lymphocytes 20%-40% (mononucleosis, hepatitis),
HIV infection, sepsis, steroid
use
leukemia

Chronic inflammatory disease

(including tuberculosis et Chemo-radiotherapy, tumors,
Monocytes 2%-8% similia), some viral infections, HIV
parasitic infections, leukemia,

Allergic reactions, Addison’s

Eosinophils 1%-4% disease, parasitic infections, Chemo-radiotherapy, tumors
some rare pathological forms

Splenectomy, allergic reactions,

myeloproliferative Chemo-radiotherapy, tumors,
Basophils 0.5%-1% disorders/leukemia, Varicella acute infection, severe injury
infection

Band (young
0%-3%
neutrophils) 25