Innate Immunity

and
Inflammation

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 What You Have To Know
• The components of the innate immunity

• Attributes of the innate immune responses

•The role of chemical mediators and adhesion molecules

•The sequence of inflammatory events and their role in the

initial response to invasion

•The mechanism and chemoattractive molecules involved in

phagocyte extravasation

•The meaning of opsonization and molecules involved

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 INNATE IMMUNITY
• It involves largely phagocytic cells (neutrophils and macrophages)
and provides a first line of defense.

• It deals with the diversity of pathogens that a person might

encounter in their lifetime by having available (at all times) a few
types of recognition molecules, each of which recognize a large
number of pathogens.

• It is: (1) rapid, (2) invariant, (3) nonspecific, (4) limited diversity,
and (5) the response to a particular type of microbe is not larger or
faster after repeated encounter (no memory response).

• Viruses induce interferons (IFN- and IFN-) which act to inhibit

viral replication. Bacterial lipopolysaccharide induces chemokines
and cytokines that lead to inflammation. 3
Protective Barriers

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 Physical/Chemical Barriers
• Mucous and ciliated cells work to trap microbes in upper respiratory tract and
sweep them to the the nasal or oral cavity for expulsion or swallowing (the
g.i. tract is less hospitable to microbes than the lungs)

• The cough/sneeze reflex can expel large numbers of microbes (on dust or
other particles) quickly

• The low pH of the stomach, bile salts and acids, and digestive enzymes kill
or limit growth of most microbes

• Low moisture and organic acids protect skin

• Competition from normal flora microbes protects from pathogens - long term
antibiotic treatment can compromise this and is often associated with yeast
infections because of decreased competition

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Physical Barriers

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Protective Barriers

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Intact Skin

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Mucous Membranes

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 Chemical Defenses
• Lysozyme in tears and saliva kills gram-positive bacteria
by degrading cell wall

• Complement system can lyse some microbes and promote

their phagocytosis. It is activated by some directly.

• Cytokines and chemokines are important in the induction

of inflammation.
– e.g. interferons (IFN-and ). Induced by viruses,
inhibit viral replication

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 Neutrophils
• Neutrophils are phagocytic
leukocytes that are able to
ingest and kill most
microbes in the blood. They
can migrate from blood to
tissues, if needed (described
more fully in the section on
inflammation).

• They are short lived cells

and require little or no
activation for maximum
effect, just a triggering
stimulus
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• Attracted to parasites Eosinophils
(helminths or protozoa)

• Secretes toxic basic

proteins onto parasite
surface

• Can be activated by
inflammatory mediators or
allergens with IgE

• Parasite-specific IgE
greatly enhances binding to
parasite

• Eosinophilia is an indicator 13
of parasitic infections
 Macrophages
• Monocytes migrate from
the blood and develop into
macrophages

• Many tissues have resident

macrophages and in some
tissues they are given
unique names:
Kupffer cells, liver
microglial cells, CNS
alveolar MØ, lungs

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 Macrophages
• Macrophages can be activated
by a variety of stimuli (e.g.,
bacterial lipopolysaccharide,
bacterial DNA, other bacterial
components, phagocytosis, or
T-cell derived cytokines) to
produce cytokines and to
prepare to kill microbes.
Activation of NFB (a nuclear
transcription factor required
for transcription of several
cytokines) by LPS is
illustrated here.

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 Macrophages
• Macrophage-derived cytokines (discussed further under
Inflammation) are critical in coordinating innate responses
to a variety of microbes.

• A major effector function of macrophages is phagocytosis

of particles (bacteria, yeast, etc.) or pinocytosis of viruses
and the use of a variety of mechanisms to kill then and
digest them. This will be discussed in greater detail in the
lecture on phagocytic processes.

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Professional Phagocytes - (phagocytosis
= ingestion of particles)
• Macrophages
– Kupffer cells
– Alveolar macrophages
– Microglial cells
– Osteoclasts
• Neutrophils
• Eosinophils

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Phagocyte in action

Bacteria
Lymphocyte
Macrophage

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 Recognition of Pathogens
• A variety of surface receptors on
phagocytes bind to microbial
components
• Phagocytic cells have receptors
that bind to antibody-coated
particles. In some cases (e.g.,
bacteria with capsules), binding
and phagocytosis are minimal
without antibody, complement, or
C-reactive protein, or mannose-
binding protein

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 Recognition

• Professional
phagocytes have
receptors for
complement
components that
greatly enhance
recognition and
binding of
microbes

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 Recognition
Toll-Like Receptors (TLR)

TLR

PAMPs = Pathogen-associated molecular patterns

The key components of innate immunity, pattern recognition receptors (PRRs), are
considered to act as sentinels against both invading organisms bearing pathogen-
associated molecular patterns (PAMPs) and damage-associated molecular patterns
(DAMPs).
•Pattern recognition
receptors (PRRs) e.g.
Toll-like receptors

•Pathogen-associated
molecular patters
(PAMPs) e.g.
Pathogenic and
nonpathogenic microbes

•Damage-associated
molecular pattern
molecules (DAMPs)
e.g. endogenous
molecules released from
dying host cells
molecules upon cellular
stress or tissue damage
 Phagocytosis

• After recognition of
microbial components by
receptors on the surface of
the phagocyte, the microbe
is ingested by means of
membrane invagination to
produce a phagosome. This
fuses with lysosomes which
have a low pH and
degradative enzymes to kill
the microbe in a structure
that is then termed a
phagolysosome.
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 Phagocytosis
• Opsonization with antibody generally enhances the process of
phagocytosis. However, some microbes (e.g., Salmonella,
Listeria, Mycobacteria) can withstand the phagolysosome
environment in unactivated macrophages

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 Phagocytosis
• Most bacteria are effectively retained in phagosomes and
destroyed when these fuse with lysosomes. However,
Listeria monocytogenes can escape into the cytoplasm in
unactivated macrophages. However, activation by exposure
to cytokines or LPS prevents this escape.

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 Killing
• Mechanisms include agents that are resident in
lysosomes of neutrophils and macrophages as well as
processes that are triggered by phagocytosis
(respiratory burst leading to generation of reactive
oxygen species).

• The respiratory burst generates large quantities of

toxic oxygen metabolites that are potent anti-
microbial agents. They are also often lethal to the
phagocyte and may damage surrounding tissues as
well
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Phagocytosis

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Functional Responses of Phagocytes

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 • Superoxide radicals are
produce from molecular
oxygen
• Hypochlorus acid is
produced when
Acidification myeloperoxidase catalyses a
pH 3.5-4.0 reaction between hydrogen
peroxide and chloride ions
• Lysozyme is oxygen
independent and acts to split
peptidoglycan
• Hydrogen peroxide is formed
NADPH when superoxide radicals
O2 + NADPH NADP + O2- + H+
Oxidase combine with hydrogen ions
• Cationic proteins are oxygen
independent and damage
O2- + H Superoxide
+ O2 + H202 microbial membranes
Dismutase

H202 + Myeloperoxidase
Cl- OCl- + H2O 29
 Natural Killer (NK) Cells
• Small percentage of total
lymphocytes
• Kill cells infected with certain
viruses
• Kill susceptible cancer cells
• Produce Interferon-gamma for
early anti-bacterial activity
• Activated by various macrophage
derived cytokines (IFN-alpha, IFN-
beta, IL-12) to more effectively
lyse target cells

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 Natural Killer (NK) Cells
• NK cells are lymphocytes with innate
immune functions
– Express a set of receptors for self proteins
induced by:
• Infections
• Malignant transformations
• Other stresses
– Activated NK cells perform one of two functions:
• Kill the altered self cell
• Produce cytokines that induce adaptive responses
against the altered self cell
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Natural Killer (NK) Cells

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 Natural Killer (NK) Cells
NK cell
• NK cells secrete
perforin onto the target
cell surface to create a
pore

• Granzymes are then

Perforins
released, enter the Granzymes
target cell via a perforin
pore, and induce
apoptosis
Tumor cell or
Infected cell

Note: NKT cells constitute only approximately 0.1% of all peripheral blood T cells.
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It is
involved in the development of autoimmunities, cancer, and asthma.
INFLAMMATION

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 Cause of Inflammation
•  Infections (bacterial, viral, fungal, parasitic) &
microbial toxins
•  Tissue necrosis: ischemia, trauma, physical or
chemical injury (e.g., thermal injury;
irradiation; some environmental chemicals)
•  Foreign bodies (splinters, dirt, sutures)
•  Immune reactions (hypersensitivity reactions)

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The process - extravasation
• Endothelial cells in regions
of infection, trauma, etc.
become activated and
express E- and P-selectin
and ICAM-1, which bind
to ligands on leukocytes
causing them to stop and
begin the process of
diapedesis. They squeeze
between endothelial cells,
secrete proteases to break
down the basement
membrane, then enter the
tissues.

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 LEUKOCYTE - ENDOTHELIAL ADHESION MOLECULES

Leukocyte

SLe x,a CD11a/

CD11a,b/ 47
L-Selectin CD18
CD18
VLA-4

P-Selectin VCAM-1
ICAM-2 MAdCAM-1
E-Selectin
ICAM- 40
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 Leukocyte-Endothelial Adhesion
Molecules
LEUKOCYTE ADHESION MOLECULES
2 Integrins: CD18
1 Integrins: VLA-4 (41)
L-Selectins (Lymphocytes)

ENDOTHELIAL CELL ADHESION MOLECULES

ICAM-1, VCAM-1, MAdCAM-1, P- and E-selectins

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 Leukocyte-Endothelial Adhesion
Molecules
Adhesion Molecule Name Function
Selectin E-selectin Leukocyte migration and homing
L-selectin Initial binding to endothelium
P-selectin Leukocyte migration to inflammation
site
ICAM ICAM-1, -2, -3 Homing to lymph node and migration to
inflammation site
LFA-3 Lymphocyte interactions
CD2 Lymphocyte interactions
Integrin VLA family Migration through extracellular matrix
LFA-1 Tight binding to endothelium
CR3 Tight binding to endothelium and
phagocytosis
CR4 Phagocytosis 42
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 The process - chemotaxis
• Leukocytes in tissues move toward
sites of infection by “crawling”
toward higher concentrations of
chemoattractants such as
chemokines like IL-8 or bacterial
products such as N-formyl peptides

A macrophage extends
pseudopods to ingest a
yeast cell

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ACUTE INFLAMMATION

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 ACUTE INFLAMMATION
•  The body's normal physiological response to injury or pathogen
invasion

• A protective response involving host cells, blood vessels and

proteins
–  Goals are:
 eliminate the initial cause of cell injury
 Remove necrotic cells and tissue
 Initiate the process of repair

• Also a potentially harmful process

– Components of inflammation that are capable of destroying microbes
can also injury bystander normal tissue
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 ACUTE INFLAMMATION
•  Components of the inflammatory process include white
blood cells and plasma proteins
 Normally present in the blood
The inflammatory reaction’s goal is to bring these to
the site of infection and/or tissue damage
•  Acute inflammation is induced by chemical mediators
produced by damaged host cells
Cytokines and other mediators
• Acute inflammation is normally controlled and self-
limited.

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 ACUTE INFLAMMATION
Later stages of inflammation are the acute phase responses
(APRs)
– Induced by pro-inflammatory cytokines (IL-1, TNF-α,
and IL-6)
– APR involves:
• Increased synthesis/secretion of antimicrobial
proteins from the liver
– MBP (Mannose Binding Protein)
– CRP (C-Reactive Protein)
– Complement components
• Liver acute phase proteins activate other processes
that help eliminate pathogens

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ACUTE INFLAMMATION

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(IL-8)

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Macrophages are stimulated by bacterial binding to
secrete inflammatory cytokines and lipid mediators of
inflammation such as prostaglandins and leukotrienes.
C5a, leukotriene B4, and histamine signal endothelial
cells to rapidly move P-selectin from granules to the
cell surface. Key inflammatory cytokines produced by
macrophages include Interleukin-1 (IL-1), which
activates nearby vascular endothelium to increase
expression of ICAM-1 and ICAM-2 to promote
leukocyte extravasation; chemokine IL-8, which
activates integrins Mac-1 and LFA-1 on leukocytes to
bind more strongly to vascular CAMs; Tumor
Necrosis Factor alpha (TNFa), which activates
vascular endothelium to express E-selectin and, with
IL-8, activates neutrophils to live longer and be more
phagocytic; and IL-12, which activates NK cells to
kill virus-infected cells. IL-1, IL-6, and TNFa also
cause fever, a regulated increase in body temperature
that stimulates immune responses and inhibits
replication of some bacteria and viruses. IL-1, IL-6,
and TNFa also signal the liver to produce acute
phase proteins. Overproduction of TNFa in response
to systemic Gram negative bacterial infection causes
septic shock: systemic edema (fluid leaving the
circulation and entering the tissues), low blood
pressure, and disseminated coagulation that leads to
multiple organ failure and death.
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Erythrocyte sedimentation rate (ESR)- measures
the amount of acute phase proteins in the blood.
Nonspecific indicator of infection, inflammation,
autoimmune disease, or malignancy

(MBP)

Acute-phase proteins
– Fibrinogen, blood clotting
– Mannan-binding protein (MBP),
opsonization
– C-reactive protein (CRP), opsonization of
bacteria with phosphorylcholine
– Serum amyloid A (SAA) protein, self-
aggregation to form scar tissue - can also
damage healthy organs if inappropriately
deposited
– SP-A and SP-D are lung surfactant proteins
– Complement components, opsonization of
bacteria 56
 Acute Phase Response
• The acute phase response is induced primarily by
cytokines from macrophages, particularly IL-1, IL-6, and
TNF- whose production can be induced by the following:
pathogenic infection, trauma, malignant neoplasms, burns,
tissue infarction, immunologically mediated inflammatory
states, crystal-induced inflammatory states (gout),
strenuous exercise, childbirth, and marked psychological
stress. The acute phase proteins are produced mostly by
hepatocytes.

• The cytokines that cause the acute phase response can also
act in the brain to induce fever and sleep - additional
defense mechanisms.
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 Acute Phase Proteins
– Fibrinogen, blood
clotting

– Mannan-binding
protein, opsonization

– C-reactive protein,
opsonization of bacteria
with phosphorylcholine

– Serum amyloid A
protein, self-aggregation
to form scar tissue - can
also damage healthy
organs if inappropriately
deposited

– SP-A and SP-D are lung

surfactant proteins 58
 IFN-

IL-6

IL-8
IFN-

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 Type-I Interferons
Antiviral action

Antiviral action
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Type-I Interferons

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 Cytokines of the Innate Response
Cytokine Secreted by Targets and Effects
IL-1 Macrophages Induces fever (pyrogen), inflammation, acute phase proteins, and the
production of IL-2 and IL-2 receptors
IL-6 Macrophages Induces acute phase proteins; influences humoral immunity
IL-8 Macrophages Chemoattraction and extravasation of neutrophils Promotes
neutrophils to live longer and be more phagocytic
IL-12 Macrophages Activates NK cells
Dendritic cells
IL-15 Macrophages Induces CD8+ T-cells and NK cells cytolytic activity
IL-18 Macrophages Induces IFN-gamma production and NK cells cytolytic activity
TNF- Macrophages Induces inflammation, acute phase proteins, cell death in many cell
types
IFN- Macrophages Antiviral factor; activates NK cells

IFN- Fibroblasts Antiviral factor; activates NK cells

IFN- NK cells Activates Macrophages

 Chronic Inflammation
If the condition causing acute inflammation is not resolved, the inflammation may
pass to a longer term chronic phase. Also, some pathogens by their nature tend to
directly provoke chronic rather than acute inflammation. Many of the features of
acute inflammation continue as the inflammation becomes chronic, including
increased blood flow and increased capillary permeability. Accumulation of white
blood cells also continues, but the composition of the cells changes.

Neutrophils quickly enter the infected tissue, and these short-lived cells
predominate initially. However, soon macrophages and lymphocytes begin to be
recruited. The sequence by which they bind to cell adhesion molecules and pass
through the endothelium. Thus, the primary cells of chronic inflammation are
macrophages and lymphocytes.

Macrophages live far longer than neutrophils. As their name suggests,

macrophages phagocytize pathogens and other material at the site of the
inflammation. Because they are long-lived, indigestible material may remain
inside macrophages in vesicles for long periods. As described previously,
macrophages are important secretory cells releasing inflammatory
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cytokines/chemokines, growth factors, and a variety of other proteins.
 Chronic Inflammation
Inflammation does not always resolve in a short period of time; it
may last indefinitely, contributing to numerous other problems.

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 Chronic Inflammation
• Infections can cause chronic inflammation
– Continual microbial invasion can induce chronic
inflammation
• Gum disease
• Unhealed wounds
• Failure of tolerance mechanisms to gut microbes
– Some microbes are ineffectively cleared,
inducing chronic inflammation
• Certain fungal and mycobacterial infections

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 Chronic Inflammation
• There are non-infectious causes of chronic
inflammation
– Damage-associated molecular patterns (DAMPs)
• Self-Antigen released by certain conditions associated
with damage
– Tumors
– Autoimmune diseases
– Atherosclerosis and heart disease
– Obesity
• Each capable of inducing long-term inflammation

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 Causes of Chronic Inflammation
•  Persistent injury or infection
–  Ulcer, tuberculosis
•  Prolonged exposure to a toxic agent
–  Pulmonary silicosis (silica in the lung)
•  Perpetuating immune reaction that results in tissue
damage and inflammation

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Chronic Inflammatory Diseases

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Chronic Inflammatory Diseases
• Acne • Obesity
• Allergies • Cardiovascular disease
• Asthma • Alzheimer's
• Celiac disease • Parkinson’s
• Crohn’s disease • Autism
• Ulcerative colitis • Mental illness
• Chronic pain • Diabetes Type I and II
• Lyme Asthma • Cancer
• Graves’ disease • Chronic Infections
• Rheumatoid arthritis • Chemical sensitivity
• Irritablebowel • Osteoporosis
syndrome • Chronic fatigue
• Gluten sensitivity • Fibromyalgia
• Insulin resistance • Infertility
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 Acute and Chronic Inflammation

Feature Acute Chronic

Onset Fast, minutes or hours Slow, days or weeks
Cellular Infiltrate Mainly Neutrophils Macrophages/Lymphocytes
Tissue injury, fibrosis Usually mild and self-limited Often severe and progressive
Local and systemic signs Prominent Less prominent, maybe subtle

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 Summary
• The innate immunity is a barrier system consisting of anatomic, physiologic,
phagocytic, or inflammatory components.

•The innate immunity has limited specificity and diversity, and is not enhanced by
repeated exposures.

•Phagocytes (the first neutrophils and then monocytes) will extravasate into the area
of injury by (1) rolling, (2) activation with chemoattractants, (3) arrest/adhesion, and
(4) transendothelial migration.

•Important chemoattractants include IL-8, C5a, leukotriene B4, and N-formyl

peptides.

•L-selectins on lymphocytes bind to addressins whereas integrins on the phagocytes

bind to ICAMs on endothelia to mediate adhesion and then reach the underlying
tissue.

•Once through the endothelium, phagocytes are attracted to the area of injury 71
by
chemokines.
 Summary
•L-selectin is constitutively expressed by leukocytes, whereas E- and P-selectin
must be induced on endothelial cells

•Integrin molecules such as α4β7 are important for leukocyte homing

•Phagocytosis involves: the formation of pseudopodia that trap particulate material

in a phagosome; addition of lysosomes to cause intracellular digestion within the
phagolysosome; and exocytosis of digested materials.

•Opsonization is the coating of foreign particles with proteins such as acute phase
proteins to enhance phagocytosis.

•Ingestion is accompanied by a respiratory burst, which generates the toxic

metabolites necessary to destroy intracellular materials.

•NADPH oxidase produces toxic oxygen radicals, and myeloperoxidase generates

hypochlorite.

•Lysosomal contents (lysozyme, defensins, lactoferin, and hydrolytic enzymes)

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are
also toxic to ingested material.