Innate Immunity (Chapter 2)

REVIEW from previous classes:

• Most organisms that are encountered daily do not cause disease; they
are detected and destroyed within hours by the mechanisms of innate
immunity.

• Only if the pathogen escapes or overwhelms the mechanisms of innate

immunity, the pathogen causes diseases, and is later recognized by the
mechanisms of adaptive immunity.

• Mechanisms of innate immunity are fixed, non-specific, inherited

from our parents, and are an essential prerequisite of adaptive
immunity.

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 Time course of primary infection in
healthy individuals, and people who
lack immune or adaptive immunity

In the absence of innate immunity, there

is also no adaptive immunity; patients
after severe burn accidents, where the
innate immunity is destroyed (skin
burned) have a high mortality rate

Infection is cleared from the body

by combined actions of innate and
adaptive immunity 2
 There are four classes of pathogens and they
infect the body through different routes

• Four classes of pathogen: Bacteria

Viruses
Fungi
Parasites
• Routes of infection: External epithelia (skin)
Internal epithelia (mucosal surfaces)

• Pathogens can replicate: Inside the cell (intracellularly)

In spaces between human cells
(extracellularly)
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Pathogens can replicate in different compartments of the body

Accessible to antibodies Killed within the infected cell

All pathogens have an extracellular stage in their life. Fore some, all life stages
are extracellular, whereas others can also grow and replicate within the cells –
intracellularly.
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 The innate (non-specific) immunity
has three components
1. Anatomical barriers: Skin, gut, lungs, eyes/nose

2. Cellular components: neutrophils and macrophages (killing

engulfed bacteria and viruses), eosinophils (killing multi-cellular
parasites), and NK cells (killing viruses and cancer cells)
3. Chemical components - soluble (released) molecules:  Cytokines
and chemokines, leukotriens and prostaglandins, complement
proteins, protease inhibitors, acute phase proteins
In a way, the immune system works best if it does not have to work at
all. This means if the pathogen is not able to enter the body, no
immune response needs to be mounted, and the person stays healthy.

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 I. The anatomical barriers of innate immunity
• The surface of human body is protected by a continuous covering of
epithelial cells, which form a physical barrier between the body and the
external environment.
Skin (provides just a physical barrier)
• Epithelium
Mucosal surfaces (epithelial linings of the respiratory,
GI, and urogenital tracts; provide communication between
body and environment; secrete mucus)
• Mucus: Thick fluid covering mucosal epithelium, and preventing pathogens
from adhering to the epithelium; contains defensins, lysozyme and protease
inhibitors. (Patients with cystic fibrosis have defective mucus secretion 
they develop frequent lung infections).
Mucus protects e.g., the stomach, which contains HCl strong enough that
could dissolve the metal Zn. The reason why our stomach is not destroyed by
the acids is the mucus; without it, our body would digest itself.

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 Epithelium is protected also by a flora of
nonpathogenic microorganisms (commensal species)

• Commensal species are nonpathogenic microorganisms that protect

epithelium by competing with pathogens for nutrients and attachment sites
(commensal = “they eat at the same table”)
• There are about 500 microbial species living in healthy human gut
• They inhibit colonization by pathogens, enhance nutrition by processing
digested food and making some vitamins, and produce antimicrobial
peptides that protect from colonization by pathogens.
• After antibiotic treatment, the nonpathogenic flora is killed, which can
result in re-infection with other pathogens (Acidophilus therapy after
antibiotic treatment facilitates re-colonization by commensal species)

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Antibiotic treatments disrupt the natural ecology of the colon

This may
cause
inflammation
and bleeding

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 II. Cellular components of innate immunity
1. Macrophages – Tissue macrophages and newly recruited monocytes, which
differentiate into macrophages; reside in tissues; usually the first cells that
come into contact with the pathogen. Function in phagocytosis and intracellular
killing of microorganisms. Contribute to tissue repair and act as antigen-
presenting cells, which are required for the induction of specific immune
responses.
2. Neutrophils – Polymorphonuclear leukocytes (PMNs) are the first cells that
are recruited to the site of infection or injury. They phagocytose invading
organisms and kill them intracellularly. In addition, PMNs contribute to tissue
damage that occurs during inflammation.
3. Natural killer (NK) cells - nonspecifically kill virus infected and tumor cells.
These cells are not part of the inflammatory response but they are important in
nonspecific immunity to viral infections and tumor surveillance.
4. Eosinophils – have proteins in granules that are effective in killing certain
parasites.

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 Macrophages
• Terminally differentiated cells that reside in
tissues.
• They differentiate from monocytes, which
circulate in the blood.
• The primary function of the macrophage is to
engulf, or phagocytose the pathogen, and
then release cytokines that activate other cells
of the immune system (neutrophils, T cells).

PHAGOCYTOSIS

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Phagocytosis of pathogen by macrophages
1. Macrophage binds the pathogen through specific
receptors expressed on the macrophage; CD14 and
Toll receptors (TLR)
2. Macrophage engulfs the pathogen and degrades it in
phagolysosomes (vesicles formed by fusion of
lysosomes and phagosomes, and filled with
proteases).
3. The activated macrophage 1) releases pro-
inflammatory cytokines (IL-1, IL-8 and TNF) that
attract neutrophils from circulation (to help the
macrophages with phagocytosis), and 2) presents
the digested pathogen as MHC class II molecules to
naive T cells to activate adaptive immunity

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Phagocytosis of pathogen by macrophage

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 Neutrophils
• The most abundant leukocytes
• One of the first cells recruited to the site of injury or infection
• Short-lived, terminally differentiated, undergo apoptosis within 24 hours
• Their main function is to phagocytose the pathogen and digest it

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 Role of neutrophils in vasodilation

Normal blood flow Vasodilation and neutrophil extravasation

• During inflammatory response, the endothelial cells that make up the walls of blood
vessels contract.
• Since these blood vessels get larger in diameter as a result of this, the process is called
vasodilation.
• Vasodilation increases the space between the endothelial cells resulting in increased
capillary permeability.
• Consequently, neutrophils can squeeze through the spaces between the endothelial cells
(= diapedesis or extravasation), and migrate to the infected (or injured) tissues.
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Eosinophils and Basophils
Eosinophils:
Involved in allergic
reactions and protection
against parasites

Basophils:
The least abundant
leukocytes, involved in
allergic reactions and
protection against
parasites

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 III. Chemical components of innate immunity

• Complement Proteins
• Protease Inhibitors (Serpins)
• Cytokines and Chemokines
• Lipid-Derived Mediators
– Prostaglandins - increase vasodilation, serve as chemoattractants for
neutrophils
– Leukotrienes - increase smooth muscle contraction, serve as
chemoattractant for neutrophils
• Acute Phase Proteins
– The majority of these proteins are synthesized by the liver.
C-reactive protein (CRP)

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 Complement proteins
• Complement is a system of plasma proteins that mark
pathogens for destruction
• Complement proteins have proteolytic function (they cleave
other proteins). They coat pathogens and form pores on
pathogen membranes, causing lysis of pathogens. Complement
fragments adhere to pathogen surface (opsonization) and
promote phagocytosis and microbial killing by leukocytes.
• Discovered as components of plasma that augments the
opsonization of bacteria by antibodies and allows antibodies to
kill some bacteria. This activity was said to ‘complement' the
antibacterial activity of antibody, hence the name.

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Complement
fixation

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Anaphylatoxins: Small
complement fragments
(C3a, C5a, etc) released
during complement
activation, induce
recruitment of
inflammatory cells and
local inflammation

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 Cytokines (= Interleukins)
• Small (MW~10-20 kD) secreted proteins, which regulate
behavior of other cells; they mediate and regulate immunity,
inflammation, cell growth and differentiation, and
hematopoiesis.
• They act by binding to specific membrane receptors, which then
signal the cell via second messengers, often tyrosine kinases, to
alter its behavior (gene expression).
• The main classes of cytokines:
– Pro-inflammatory: Interleukin (IL)-1, IL-6,
Tumor Necrosis Factor (TNF)
– Anti-inflammatory: IL-10
– Growth factors: IL-4
– Interferons: IFNg
– Chemokines: IL-8 20
chemokines
Function of cytokines

• autocrine function
– same cell that produced cytokine
• paracrine function
– cells nearby
• endocrine function
– distributed by circulatory system to distant target cells

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 Pro-inflammatory cytokines
• Low MW proteins that regulate function of other cells and
organs
• Released by inflammatory cells; induce inflammation: IL-8,
IL-1, IL-6, TNF, interferons
• Involved in many inflammatory diseases (asthma, chronic
lung disease, sepsis, myocardial infarction, arthritis, cancers,
etc)
• Targets of many anti-inflammatory drugs (steroids-
glucocorticoids, advil, aspirin)

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Role of pro-inflammatory cytokines in health and disease

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 Anti-inflammatory cytokines
• Inflammation often results from increased production of pro-
inflammatory cytokines, and decreased production of anti-
inflammatory cytokines
• IL-10 is the most potent anti-inflammatory cytokine

• They are produced by macrophages and by regulatory T cells – Treg

(CD25+)
• Synthesis and release of IL-10 is decreased in chronic inflammatory
diseases such as chronic lung injury or bronchopulmonary dysplasia
(BPD)
• The anti-inflammatory cytokines such as IL-10 represent one of the
most promising new anti-inflammatory therapies (inflammatory
bowel disease = Crohn’s disease; BPD)
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 Chemokines
• Low MW proteins that attract (recruit) leukocytes to the inflamed
tissues. Central role in the inflammatory response.
• Produced by activated inflammatory cells (neutrophils and
macrophages) in response to injury or infection.
• One of the most pro-inflammatory chemokines is interleukin-8 (IL-8),
which induces neutrophil migration and recruitment
• Role also in cancer progression; metastases and angiogenesis

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 Cytokines induced by
activated macrophages
in response to
stimulation with
lipopolysaccharide

= IL-8

= endocrine function

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LOCAL SYSTEMIC
Pro-inflammatory cytokine tumor
necrosis factor (TNF) induced by
activated macrophages induces
protection at the local level, but when
released systematically, it can cause
systemic inflammation and sepsis

Local effect: TNF activates neutrophils that

engulf the pathogen and then undergo
apoptosis  resolution of inflammation

Systemic effect: TNF can induce systemic

inflammatory response, and uncontrollable
inflammation resulting in sepsis (and in
30% of cases, in death)

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 Sepsis and Septic Shock
• Sepsis is a complex illness where the body “over-reacts” to an infection.
Normally, the body’s response to an infection is targeted to the site of the
infection. With sepsis, the immune system sets off a chain reaction to fight
the infection. The body’s response, instead of being localized to the site of
infection, causes symptoms to occur throughout the body (this is known as
a systemic response).
• As a result of this systemic response, a patient with sepsis often has a
fever, and a faster than normal heart rate and breathing rate. Because
pathogens may be found in the blood of patients with sepsis, doctors and
nurses sometimes refer to the condition as a “blood infection”.
• In some patients, the systemic response to infection may spin out of
control, throwing off the body’s state of balance and damaging one or
more vital organs (heart, lungs, kidneys, or liver). This systemic response
or “over-reaction” to the infection is called severe sepsis or septic shock.

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 Pro-inflammatory cytokines produced by
activated macrophages can result in sepsis

• Pro-inflammatory cytokines released by activated macrophages and

neutrophils can cause systemic inflammation and sepsis (uncontrollable
systemic inflammation that is mediated by blood cells and eventually spreads
to all organs, causing multiple organ failure).
• Septic shock is the most common cause of death in the intensive care
units. Between 33% and 50% of patients who develop severe sepsis die from
the condition (~ 100,000 patients die from sepsis in the U.S. each year).
• There is no effective and specific therapy for sepsis  subject of intensive
research; current treatment includes anti-inflammatory therapy
(glucocorticoids) and monoclonal antibodies against TNF and other pro-
inflammatory cytokines.

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 COVID-19 cytokine storm
• COVID-19 infection can cause a cytokine COVID-19
storm - overproduction of pro-inflammatory
cytokines, such as TNF-α, IL-1, IL-6, and IL-8,
which can result in increased migration of
activated immune cells into the lungs, resulting
in lung inflammation and respiratory distress
called Acute Respiratory Distress
Syndrome, which often leads to death.
• The cytokine storm can also affect other vital
organs (heart, brain, kidney), resulting in
increased mortality.

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 Summary – Innate Immunity
• Most infections are cleared by the mechanisms of innate immunity;
prerequisite for adaptive immunity
• Focused on antigen destruction by phagocytosis, by neutrophils and
macrophages
• Has three components:
 Anatomical barriers: Skin, gut, lungs, eyes/nose
 Cellular components: neutrophils and macrophages (killing engulfed
bacteria and viruses), eosinophils (killing multi-cellular parasites), and NK
cells (killing viruses and cancer cells)
 Chemical components - soluble (released) molecules: Cytokines and
chemokines, leukotriens and prostaglandins, complement proteins, protease
inhibitors, acute phase proteins
• Sepsis - complex illness where the body “over-reacts” to infection.
Characterized by increased production of pro-inflammatory cytokines, such
as TNF. Can lead to septic shock, which is the most common cause of death
in the intensive care units; there is no specific and effective treatment for
septic shock

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