Inflammatory Process.

Mediators of
Inflammation.
Lymphocyte
Recirculation.

What is inflammation?
In Latin: inflammare (to set on fire).
It is body defence reaction in order to eliminate or limit the
spread of injurious agent as well as to remove necrotic cells
and tissues. Initiates the proces of repair
Also a potentially harmful process. Components of
inflammation that are capable of destroying microbes can
also injury normal tissue

Excess inflammatory reactions

Inappropriate inflammatory response when there
are no foreign substances to fight off leads to
autoimmunity
Inflammatory process must be tightly regulated by
immune system to avoid excessive damage normal
tissue

Inflammation is
normally
controlled and
self-limited

Components of the inflammatory proces include white blood cells

and plasma proteins
Normally present in the blood
The inflammatory reactions goal is to bring these to the site of
infection and/or tissue damage
Inflammation is induces by chemical mediators produced by
damaged host cells (Cytokines and other mediators)

Types of Inflammation

The acute inflammatory response involves both localized

(redness, swelling, heat, pain) and systemic responses.

Signs of acute inflammation

Heat

Redness

Swelling

Pain

Loss of Function

The local reactions are described as the cardinal signs and symptoms of
inflammation

Signs of acute inflammation

(Local manifestations of acute inflammation)
Redness (Rubor) & Heat (Calor) :
This is because the capillaries are filled up with more blood
than usual (blood vessel dilation).
(The increased volume heats the tissue and causes it to redden)

Mediators promote the vasodilation

oHistamine
oSerotonin
oBradykinin
oProstaglandin
1x increase in arteriole diameter yields a 4x increase in
blood flow

Signs of acute inflammation (Local manifestations of

acute inflammation) cont.
Swelling (Tumor) / Edema - caused by an accumulation of fluid
Edema fluid varies with the stage of inflammation
Initially vessel permeability is only slightly altered and no cells
or proteins escape and the fluid is mainly water and dissolved
electrolytes.
As capillary permeability increases and plasma proteins escape
the extravascular fluid becomes cloudy and more viscous. This is
called exudate (contains a large amount of leukocytes)
extravasation of leukocytes

Signs of acute inflammation (Local manifestations of

acute inflammation) cont.
Pain (Dolor)The pain associated with inflammation, results in part from the
distortion of tissues caused by edema.
Chemicals mediators (bradykinin, serotonin, and the
prostaglandins) that stimulate nerve endings are released, making
the area much more sensitive.
Physical trauma may irritate pain receptors.
Loss of function (Functio laesa ) may occur due to pain
causing reflex guarding or muscle spasm.
Loss of function may result from pain that inhibits mobility or
from severe swelling that prevents movement in the area.

The Inflammatory response consist of

a vascular and cellular reaction

Vascular changes
o vasoconstriction (momentary constriction of small blood vessels in the
area) to decrease blood flow (very brief - first seconds after tissue injury).

o vasodilation - within minutes after tissue injury,

there is an increase in vascular diameter resulting
in an increase in the volume of blood in the area
and a reduction in the flow of blood.
/The increased blood volume heats the tissue and
causes it to redden/.
o Vascular permeability also increases, leading to
leakage of fluid from the blood vessels,
particularly at postcapillary venules. This results
in an accumulation of fluid (swelling/edema)
o emigration of leukocytes from microcirculation

The Inflammatory Process

Cellular Reaction
Two types of leukocytes participate in the acute
inflammatory response - the neutrophils and monocytes.
1st line of defense: neutrophils
o Within a few hours of the onset of vascular changes neutrofils adhere to
the endothelial cells, and migrate out of the blood into the tissue spaces.
o Neutrophils phagocytose invading pathogens and release mediators that
contribute to the inflammatory process
o Release chemokines that help to recruit macrophages - macrophage
inflammatory proteins (MIP-1 and MIP-1)

The Inflammatory Process

Cellular Reaction
2nd line of defense: monocytes /macrophages
o Macrophages arrive about 5-6 hours after an inflammatory
response begins
o Macrophages exhibit increased phagocytosis and increased release
of mediators and cytokines
o Activated tissue macrophages secrete IL-1, TNF- and IL-6.

The acute inflammatory response involves both localized

(redness, swelling, heat, pain) and systemic responses.

The Systemic Acute-phase Response

Acute inflammatory responses that are not
restricted to the initiation site.
This response is marked by
Hormone production (ACTH,
hydrocortisone)
Increased production of white blood
cells (leukocytosis)
Synthesis of acute-phase proteins in the
liver
The increase in body temperature

Acute Inflammatory Process

Acute Inflammation Process

Leukocyte extravasation
Leukocytes must be able to get to the site of injury
from their usual location in the blood
The process of leukocyte movement from the blood
to the tissues through the blood vessels is known as
extravasation.

 As an inflammatory response develops, various cytokines and other

inflammatory mediators act upon the local blood vessels, inducing
increased expression of endothelial Cell Adhesion Molecules (CAM).

The vascular endothelium is then said to be activated, or

inflamed.

The leukocyte-extravasation cascade

Leukocytes leave the vasculature routinely through the following sequence of events:
1. Margination and rolling
2. Activation and adhesion
3. Transmigration via diapedesis
4. Movement of leukocytes within the tissue via chemotaxis

1. Margination and Rolling

With increased vascular permeability, fluid leaves the vessel causing
leukocytes to settle-out of the central flow column and marginate
along the endothelial surface.
Endothelial cells and leukocytes have complementary surface
adhesion molecules which briefly stick and release causing the
leukocyte to roll along the endothelium until it eventually comes to a
stop as mutual adhesion reaches a peak

 P-selectin is the most important selectin involved in rolling.

Upon stimulation by trauma, P-selectin is rapidly surfaceexpressed on the venular endothelium, and it makes the
endothelium "sticky" to leukocytes.

E-selectin is also
responsible for slow
rolling interactions below
5mm/s and possibly the
initiation of adhesion.

A chemokine such as IL-8 binds to a G-protein-linked

receptor on the neutrophil, triggering an activating signal.

2. Activation and adhesion

Rolling comes to a stop and adhesion results
Interactions between integrins (leukocyte: LFA-1, Mac-1, VLA-4)
and their immunoglobulin ligands (ICAM-1, VCAM-1) expressed
on the endothelium mediate the adhesion of leukocytes. (ICAM-1
binds LFA-1/Mac-1, VCAM-1 binds VLA-4)

3. Transmigration (diapedesis)
Migration across the endothelium
Occurs after adhesion within system of venules and capillaries via
PECAM-1 (CD31) (platelet endothelial cell adhesion molecule) on
endothelial cells, neutrophils, monocytes/macrophages,
lymphocytes

Mediators of Inflammation
Derive from cells or plasma proteins
Cell-derived stored in granules and can be rapidly
secreted by exocytosis or synthesized de novo
Plasma-derived produced in liver and present in
blood as inactive precursors
Active mediators can be produced in response to
stimuli
One mediator can stimulate release of other mediator
Vary in range of cellular targets
Short-lived

Cell-derived Mediators
Mediator

Sources

Actions

Histamine

Mast cells,
basophils, platelets

Vasodilation,
vascular
permeability,
endothelial
activation

Serotonin

platelets

Vasodilation,
vascular
permeability

Prostaglandins

Mast cells,
leukocytes

Vasodilation, pain,
fever

Leukotriens

Mast cells,
leukocytes

vascular
permeability,
chemotaxis,
leukocyte adhesion
and activation

Platelet-activating

neutrophils,

Vasodilation,

Inflammatory Mediators
Platelet-activating factor (PAF)
It is generated from a lipid complex stored in cell
membranes;
It affects a variety of cell types and induces platelet
aggregation;
It activates neutrophils and is a potent eosinophil
chemoattractant;
It contributes to extravascularization of plasma proteins
and so, to edema.

Cell-derived Mediators
Mediator

Sources

Actions

Reactive oxygen
species

leukocytes

Killing of microbes,
tissue damage

Nitric oxide

Endothelium,
macrophages

Vascular smooth
muscle relaxation,
killing of microbes

Cytokines (TNF, IL1)

Macrophages,
endothelium, mast
cells

Endothelial
activation, fever,
pain, anorexia,
hypotension,
vascular resistance
(shock)

Chemokines

Leukocytes,
macrophages

Chemotaxis,
leukocyte
activation

Plasma protein-derived
mediators
Mediator

Source

Actions

Complement
products (C5a,
C3a, C4a)

Plasma (produced in
liver)

Leukocyte
chemotaxis and
activation,
vasodilation

Kinins (bradykinin) Plasma (produced in

liver)

vascular
permeability,
smooth muscle
contraction,
vasodilation, pain

Proteases
activated during
coagulation

Endothelial
activation,
leukocyte
recruitment

Plasma (produced in
liver)

Mediators of Inflammation

Role of mediators in
inflammation
Role in Inflammation

Mediators

Vasodilation

Prostaglandins, NO, Histamine

vascular permeability

Histamine, serotonin, C3a, C5a,

bradykinin, leukotrienes C4, D4, E4,
PAF, Substance P

Chemotaxis, leukocyte recruitment

and activation

TNF, IL-1, chemokines, C3a, C5a,

Leukotriene B4

Fever

IL-1, TNF, Prostaglandins

Pain

Prostaglandins, bradykinin

Tissue damage

Lysosomal enzymes of leukocytes,

reactive oxygen species, NO

The cells and mediators involved in an acute inflammatory response.

The cytokines TNF and interleukin 1

(IL1) are released first and initiate
several cascades.

Short term
friend, Long
term foe

Types of Inflammation
It is rapid response to injury or microbes and other foreign
substances that is designed to deliver leukocytes and plasma
proteins to sites of injury

1. Acute Inflammation
2. Chronic Inflammation
It is inflammation of prolonged duration (week to months to
years) in which active inflammation, tissue injury and healing
proceed simultaneously

Differences between acute and chronic

inflammation
Features

Acute inflammation

Chronic inflammation

Duration of corse

Short [days]

Long [ weeks to months]

Causative agents

Physical and chemical damages; Presistent infection; presence of

pathogen invasion
forein bodies; autoimmunity

Cardinal signs

Pain; heat, redness, swelling;

loss of function

Absent in any of cardinal signs

Fundamental cells

Neutrophils
macrophages

Lymphocytes [T and B];

macrophages; fibroblasts

Fluid Exudation
and Edema

Present

Absent

Fibrosis

Absent

Present

Angiogenesis (new Absent

vessel formation)

Present

Primary Mediators

IFN, TNF, Hydrolyzing

enzymes

Serotonin; Histamine;
prostaglandins; tromboxane

Differences between acute and chronic inflammation - Pathogenesis

Acute inflammation

Chronic inflammation

1. Increased blood flow

a. Transient vasocontriction upon
endothelial injury
b. Followed by released of cytokines
that promotes vasodilation leads to
warmness and redness of injured
area
2. Increased capillary permeability
3. Migration of neutrophils
a. Rolling
b. Adhesion
c. Diapedesis
4. Chemotaxis /movements of
neutrophils to the injurous agents/
5. Leukocytes recruitment and
activation /leukocytosis/

1. Infiltration of mononuclear
phagocyting cells
a. Macrophages /circulate as monocytes
and reach site of injury within 24-48
hours and transform. Activated by
numerous cytokines from the injured site/
b. T and B cells /Recruited and activated by
APC like macrophages and DC/
c. B cells will be become plasma cells and
produce antibodies
d. T cells will produce cytokines to activated
the B cells and also macrophages
2. Tissue destruction (due to massive
production of ROS, hydrolytic enzymes)
3. Tissue repair (Angiogenesis at the injured
sites. Formation of granulomas. Fibrosis)

Termination of acute inflammation

Inflammation declines spontaneously
Mediators of inflammation are produced in rapid bursts only while the
stimulus persists
Mediators have short half-lives and are degraded after their release.
Neutrophils also have short half-lives in tissues and die by apoptosis
within a few hours after leaving the blood

Active termination mechanisms include

Switch from pro-inflammatory leukotrienes to anti-inflammatory
lipoxins
Release of anti-inflammatory cytokines, including transforming growth
factor- (TGF-) and IL-10, from macrophages
Production of anti-inflammatory lipid mediators, called resolvins and
protectins

Lymphocyte recirculation
Lymphocyte extravasation is similar to neutrophil extravasation,
Lymphocytes continuously travel from the blood to secondary lymphoid
tissues and then return to the blood.
Lymphocytes leave blood via high endothelial venules (HEVs) (HEVs need to allow egress of naive cells from the circulation)
Lymphocyte trafficking exposes antigen to a large number of
lymphocytes

Extravasation of lymphocytes includes the same

basic steps as neutrophil extravasation but some
of the cell-adhesion molecules differ.
Activation of the integrin LFA-1, induced by chemokine
binding to the lymphocyte, leads to firm adhesion followed by
migration between the endothelial cells into the tissue

Naive Lymphocytes Recirculate

to Secondary Lymphoid Tissue
Naive T cells tend to home to secondary
lymphoid tissues through their HEV
regions.
Naive lymphocytes express L-selectin
The initial interaction involves the
homing receptor L-selectin and mucinlike cell-adhesion molecules such as
CD34 or GlyCAM-1 expressed on HEV
cells.

Naive Lymphocytes Recirculate

to Secondary Lymphoid Tissue
A naive lymphocyte is not able to mount an immune
response until it has been activated to become an
effector cell.
Activation of a naive cell occurs in specialized
microenvironments within secondary lymphoid tissue
(e.g., peripheral lymph nodes, Peyers patches,
tonsils, and spleen)

Effector and Memory Lymphocytes Adopt

Different Trafficking Patterns
A second subset of memory/effector
cells displays preferential homing to the
skin.
This subset also expresses low levels of
L-selectin but displays high levels of
cutaneous lymphocyte antigen (CLA) and
LFA-1, which bind to E-selectin and
ICAMs on dermal venules of the skin.

Chemokine are important in

controlling cell traffic to lymphoid
tissues

Naive T cells:
Express chemokine receptors CCR7 and CXCR4 which allow them to
respond to chemokines (CCL21) expressed in lymphoid tissues;
Respond to chemokines CCL18 and CCL19 produced by dendritic cells,
which is thought to direct them to the appropriate T cell areas of lymph
node where dendritic cells can present antigen to them

B cells:
Express CCR7 and use similar mechanisms to migrate into the lymphoid
tissues.
B cells also express CXCR5, which allows them to respond to CXCL13, a
chemokine produced in lymphoid follicles B cells are therefore directed
to the B cell areas of the node.

Thank you for your attention

 The duration and intensity of the local acute

inflammatory response must be carefully
regulated to control tissue damage and facilitate
the tissue-repair mechanisms that are necessary
for healing.
TGF- has been shown to play an important role
in limiting the inflammatory response. It also
promotes accumulation and proliferation of
fibroblasts and the deposition of an extracellular
matrix that is required for proper tissue repair.

Process of inflammation

The leukocyte-extravasation cascade

Leukocytes leave the vasculature routinely through the
following sequence of events:
Margination and rolling
Activation, Adhesion and transmigration
Chemotaxis

They are then free to participate in: phagocytosis and degranulation

Leukocyte-induced tissue injury

Inflammatory Mediators
Complement fragments and cytokines
It stimulates chemotaxis of neutrophils, eosinophils and
monocytes;
C3a, C5a increase vascular permeability;
Cytokines
Interleukins (IL-1, IL-6, IL-8)
Stimulates the chemotaxis, degranulation of neutrophils and
their phagocytic activity
Induce extravascularization of granulocytes
Fever
Tumor necrosis factor (TNF) and IL-8
Leukocytosis
Fever
Stimulates prostaglandins production

 As an inflammatory response develops, various cytokines and other

inflammatory mediators act upon the local blood vessels, inducing increased
expression of endothelial Cell Adhesion Molecules (CAM).
The vascular endothelium is then said to be activated, or inflamed.

The process by which leukocytes migrate in response

to a chemical signal is called chemotaxis.
Exogenous agents:
Bacterial products
Endogenous products
Components of complement
Leukotriens LTB4
Cytokines / Chemokines

C5s
IL-8

Inflammatory Mediators
Prostaglandins
Prostaglandins contribute to vasodilation, capillary
permeability, and the pain and fever that accompany
inflammation.
The stable prostaglandins (PGE1 and PGE2) induce
inflammation and potentiate the effects of histamine and
other inflammatory mediators:
They cause the dilation of precapillary arterioles (edema),
lower the blood pressure, affect the phagocytic activity of
leukocytes.
The prostaglandin thromboxane A2 promotes platelet
aggregation and vasoconstriction.

Inflammatory Mediators
Leukotrienes
The leukotrienes have been reported to affect the
permeability of the postcapillary venules, the adhesion
properties of endothelial cells, and stimulates the
chemotaxis and extravascularization of neutrophils,
eosinophils, and monocytes.

Inflammatory Mediators
Histamine
It is found in high concentration in platelets,
basophils, and mast cells.
Causes dilation and increased permeability of
capillaries (it causes dilatation of precapillary
arterioles, contraction of endothelial cells and
dilation of postcapillary venules).

Inflammatory Mediators
Platelet-activating factor (PAF)
It is generated from a lipid complex stored in cell
membranes;
It affects a variety of cell types and induces platelet
aggregation;
It activates neutrophils and is a potent eosinophil
chemoattractant;
It contributes to extravascularization of plasma proteins
and so, to edema.

Inflammatory Mediators
Plasma Proteases
The plasma proteases consist of:
Kinins
Bradykinin - causes increased capillary permeability
(implicated in hyperthermia and redness) and pain;
Clotting factors
The clotting system contributes to the vascular
phase of inflammation, mainly through fibrin
peptides that are formed during the final steps of the
clotting process.

Many of the integrins can bind to more than one ligand.

e.g.:
LFA-1 present on most lymphocytes binds to both
intercellular CAM-1 (ICAM-1) and ICAM-2, which
are expressed on endothelium;
VLA-4 binds to vascular CAM-1 (VCAM-1)
expressed on endothelium, or to fibronectin (an
extracellular matrix component)

Chronic inflammation = long duration

Duration - from several months to years
Causative agent - non-degradable pathogens that cause persistent
inflammation, persistent foreign bodies, overactive immune system
reactions, autoimmune disease - self-perpetuating immune reaction that
results in tissue damage and inflammation
Major cells involved - Macrophages, lymphocytes, plasma cells and
fibroblasts
Outcomes - the destruction of tissue by inflammatory cells, thickening
and scarring of connective tissue (fibrosis), angiogenesis (new vessel
formation) death of cells or tissues (necrosis)

 High endothelial venules (HEV)

Constitutively present in secondary lymphoid tissue
Need to allow egress of naive cells from the circulation
Post-capillary venules (PCV)
Present in non-lymphoid tissues
Injury and inflammation alters morphology to resemble
HEV
Need to allow egress of effector/memory cells to sites of
infection

Effector and Memory Lymphocytes Adopt

Different Trafficking Patterns

Unlike naive lymphocytes, subsets of the

memory and effector populations
exhibit tissue-selective homing behavior.
A mucosal homing subset of
memory/effector cells has high levels of
the integrins LPAM-1and LFA-1, which
bind to MAdCAM and various ICAMs on
intestinal lamina propria venules

Inflammation-induced lymphocyte
recirculation
During the initiation of an adaptive immune response, a
large number of naive lymphocytes are recruited
specifically to the LNs draining the site of immunization
or infection.
This process facilitates the search of rare antigen
specific lymphocytes by increasing the efficiency of
screening cognate lymphocytes within the secondary
lymphoid organs.
Increased input of naive lymphocytes to the LN is in part
mediated by a rapid increase in the number of HEVs.