Mediators of inflammation and

immune reaction –1
Bradykinin,Nitric oxide,
Neuropeptides and cytokines
SAITOTI S.S.M
 Bradykinin-Biosynthesis
• Bradykinin and lysyl bradykinin (kallidin) are
active peptides formed by proteolytic
cleavage of circulating proteins (kininogens)
through a protease cascade pathway
• Bradykinin is formed from high-molecular-
weight kininogen in plasma by serine
protease kallikrein (fig. 13.12)

• Kininogen is a plasma α-globulin that exists

in high and low molecular weight forms
• Kallikrein is derived from the inactive
precursor prekallikrein by the action of
Hageman factor (factor XII)

• Factor XII is activated by contact with

negatively charged surfaces such as
collagen, and bacterial lipopolysaccharides
• Factor XII, prekallikrein and the kininogens leak out of
the vessels during inflammation due to increased
vascular permeability

• In addition to plasma kallikrein there are other kinin-

generating isoenzymes found in:
– Pancreas
– Salivary gland
– Colon
– Skin

• These tissue kallikreins act on both high- and low –

molecular-weight kininogens and generate mainly
kallidin, a peptide with actions similar to those of
bradykinin
Bradykinin–metabolism and
inactivation
• Bradykinin and related kinins are inactivated by
specific enzymes known as kininases

• Kininase II, a peptidyl dipeptidase inactivates

kinins by removing the two C-terminal amino
acids

• Kininase II is bound to the luminal surface of

the endothelial cells and is identical to
angiotensin-converting enzyme (ACE)
• Kinins are aslo metabolised by various less
specific peptidases including serum carboxy
peptidase, which removes the C-terminal
arginine generating des-Arg9-bradykinin, a
specifi agonist at bradykinin receptor B1
 Bradykinin - receptors
• There are two bradykinin receptors
designated as B1 and B2
• Both are G-protein-coupled receptors and
mediate very similar effects
• B1 receptors are normally expressed at very
low levels but strongly induced in inflammed
or damaged tissue by cytokines such as IL-1

• A number of selective peptide antagonists are

known for the bradykinin receptors
• It is likely that B1 receptors play a significant
role in inflammation and hyperalgesia and
there is interest in developing antagonists for
use in cough and neurological disorders

• B2 receptors are constitutively present in

normal cells and are activated by bradykinin
and kallidin
Bradykinin – pharmacological
actions
• Vasodilatation (largely dependent on
endothelial cell nitric oxide and PGI2)
• Increased vascular permeability
• Stimulation of pain nerve endings

• Stimulation of epithelial ion transport and fluid

secretion in airways and GIT
• Contraction of intestinal and uterine smooth
muscle
Nitric oxide as a mediator of
inflammation
• iNOS is the chief isoform relevant to
inflammation
• Virtually all inflammatory cells express iNOS in
response to cytokine stimulation
• Nitric oxide probably has a net pro-inflammatory
effect
– Increases vascular permeability and prostaglandin
production
– Is a potent vasodilator
• Some other properties of nitric oxide may be
anti-inflammatory (e.g. eNO inhibition of
adhesion of neutrophils and platelets and
platelet aggregation)

• Nitric oxide also has cytotoxic actions, killing

bacteria, fungi, viruses, and metazoa
parasites, enhancing local defence
mechanisms

• Inhibitors of iNOS are under investigation for

treatment of inflammatory conditions
 Neuropeptides
• Neuropeptides released from sensory neurons
cause neurogenic inflammation

• Substance P,neurokinin A and calcitonin gene-

related peptide (CGRP) are the main peptides
involved

• Substance P and neurokinin A (members of the

tachykinin family) act on mast cells, releasing
histamine and other mediators producing smooth
muscle contraction and mucus secretion
• CGRP is a potent vasodilator

• Neurogenic inflammation has been

implicated in the pathogenesis of several
inflammatory conditions including:
– Delayed phase of asthma

– Allergic rhinitis

– Inflammatory bowel disease

– Some types of arthritis

 Cytokines
• Cytokine is an all-purpose functional term
applied to protein or polypeptide mediators
synthesised and released by cells of the
immune system during inflammation

• More than 100 cytokines have been identified

and the super family is generally regarded as
comprising:
– Interleukins
– Chemokines
– Interferons
– Colony-stimulating factors
– Growth factors and tumour necrosis factors (TNFs)
• Cytokines act locally by autocrine or paracrine
mechanisms
• On the target cell, cytokines bind to and
activate specific high-affinity receptors that in
most cases are up-regulated during
inflammation

• Except for chemokines,which act on G-

protein-coupled receptors,most cytokines act
on kinase-linked receptors regulating
phosphorylation cascades that affect gene
expression
• Some cytokines amplify inflammation by
inducing formation of other inflammatory
mediators

• Other cytokines induce receptors for other

cytokines on their target cell, or engage in
synergistic or antagonistic inter-actions with
other cytokines

• Various systems of classifying cytokines can

be found in the literature
• For purpose of this presentation,cytokines
can be divided into:

• Those involved in the induction of the

immune response

• Pro-inflammatory and Anti-inflammatory

cytokines involved in the effector phase of the
immune/inflammatory response
• Pro-inflammatory cytokines
– Participate in acute and chronic inflammatory
reactions as well as repair and resolution

– The primary pro-inflammatory cytokines are

TNF-α and IL-1

– IL-1 comprises a family of three cytokines (IL-

1α, IL-1β, and IL-1ra)

– Mixtures of these are released from

macrophages and many other cells during
inflammation
– Pro-inflammatory cytokines can initiate the
synthesis and release of a cascade of
secondary cytokines including chemokines

– Various cytokine growth factors (e.g.

Platelet derived growth factor) are crucial
to the repair process and are implicated in
chronic inflammation
• Anti-inflammatory cytokines
– These inhibit aspects of the inflammatory
reaction

– They include: transforming growth factor

(TGF-β), IL-4, IL-10, and IL-13

– They inhibit chemokine production

– Anti-inflammatory interleukins can inhibit

responses driven by Th1 cells, whose
inappropriate activation is involved in the
pathogenesis of several diseases
• Chemokines
– Defined as chemoattractant cytokines that
control the migration of leukocytes

– Many have other actions (e.g. causing mast cells

to degranulate or promote angiogenesis)

– More than 40 chemokines have been identified

– Can be conveniently distinguished by

considering whether key cysteine residues in the
polypeptide chain are adjacent (C-C) or
separated by another residue (C-X-C)
– The C-X-C chemokines (e.g. IL-8) act on
neutrophils and are predominantly involved in
acute inflammatory responses

– The C-C chemokines (e.g. monocyte

chemoattractant protein [MCP-1] and RANTES)
act on monocytes, eosinophils and other cells,
and are involved predominantly in chronic
inflammatory responses

– Chemokines act through G-protein-coupled

receptors, and alteration or inappropriate
expression of these is implicated in:Multiple
Scelerosis,Cancer,Rheumatoid artthritis and
some Cardiovascular diseases
– Some types of virus can exploit the
chemokine system and subvert the hosts
defences

– The HIV virus is responsible for the most

audacious (daring) exploitation of the host
chemokine system

– HIV has a protein (gp 120) in its envelope

that recognises and binds T-cell receptors
for CD4 and a chemokine coreceptor that
allows it to penetrate the T-cell
• Interferons
– There are three classes of interferons (IFN-α,
INF-β, and INF-γ
– IFN-α is a family of approximately 20 proteins
with similar activities
– IFN-α and INF-β have antiviral activity
– INF- α also has some anti-tumour action
– Both (IFN-α and INF-β) are released from virus-
infected cells and activate anti-viral activity
mechanisms in neighbouring cells
– INF-γ has a role in induction of Th1 responses
• Clinical use of interferons
IFN-α is used in the treatment of chronic hepatitis
B and C, and has some action against herpes
zoster
– IFN-α is also used in the prevention of the
common cold

– INF-β is used in some patients with Multiple

Scelerosis

– INF-γ is used in chronic granulomatous disease

in conjuction with anti-bacterial drugs
 Figure 13-3 The armed CD8+ T cells also synthesise and
express IL-2 receptors and release IL-2, which stimulates the
cells by autocrine action to proliferate and give rise to
cytotoxic T cells. These can kill virally infected cells. IL-2
secreted by CD4+ cells also plays a part in stimulating CD8+
cells to proliferate. Note that the 'effector phase' depicted
above relates to the 'protective' action of the immune
response. When the response is inappropriately deployed-as
in chronic inflammatory conditions such as rheumatoid
arthritis-the Th1 component of the immune response is
dominant and the activated macrophages (mφ) release
IL-1 and tumour necrosis factor-α, which in turn trigger
the release of the chemokines and inflammatory cytokines
that play a major role in the pathology of the disease
ANY QUESTION?????????