IMM250

Immunity
&
Infection

Dr. Ray Amith

Office: MSB 7208
Email: [email protected]
Lecture 3: Part 1 • Cells of the innate immune system
• Phagocytes
• Degranulating cells
• NK cells
Innate Immunity:
• Secreted (soluble) mediators of the innate
Cellular and Soluble immune system

Mediators • Cytokines and chemokines

• Complement system
 Innate and Adaptive Immunity
E. coli

Innate (Natural) Immunity Adaptive (Acquired) Immunity

(non-specific responses) (specific responses)

1st line 2nd line 3rd line

1. Skin 1. Innate immune cells

Specialized Lymphocytes
2. Mucous 2. Inflammation
1. B cells (produce antibodies)
membranes &
2. T cells
secretions 3. Complement
• Helper T cells (CD4)
• Killer T cells (CD8)
3. Normal flora 4. Antimicrobial
substances
Immune Cells in a Human Blood Smear
 All immune cells are formed from the hematopoietic stem cell

Self-renewing, multipotent

• Hematopoiesis: forms all the

blood cells in our body (both
erythrocytes and
leukocytes).

• It occurs during embryonic

development and
throughout adulthood to
produce and replenish our
blood cells.
After entering tissues, pathogens are recognized by PRRs expressed
on phagocytic sensor cells: macrophages and dendritic cells

Macrophages and dendritic cells

express signalling receptors and
receptors that induce phagocytosis. Macrophage Dendritic cell

Present at the front-lines of the host prior to infection

Some receptors directly recognize microbial

components, some recognize host molecules
(called opsonins) deposited on pathogens.

Image: top, Icons from Biorender.com,

bottom: L. Clemenza
 Opsonins facilitate phagocytosis

• Opsonization prepares a particle for phagocytosis by coating

it with host molecules called opsonins.

• Phagocytes have receptors that recognize opsonins and can

mediate internalization.

• The main opsonins are antibodies and complement proteins

(e.g. C3b) that bind to the microbial surface.

• Antibody- and C3b-coated pathogens are recognized by Fc

receptors and complement receptors, respectively, expressed
on phagocytic cells. Phagocyte

• Other molecules that act as opsonins are produced during

the acute phase response (e.g. C-reactive protein, MBL)

L. Clemenza. Created with Biorender. com

igated. In areas where parasites are less of a (b) Macrophage
em, eosinophils are better appreciated as con-
asthma and allergy symptoms. Like neutro- Pseudopodia
phils may also secrete cytokines that regulate Phagosomes
phocytes, thereby influencing theMacrophages
adaptive Lysosome
Lysosome
ponse.
ponse. Phagolysosome
Phagolysosome
ls are nonphagocytic granulocytes
ls are nonphagocytic granulocytes (see (see
that
that contain
contain large
large basophilic
basophilic granules
granules that
that (c)
(b)
(c) Dendritic
Dendritic cell
cell
standard
standard H&EH&E staining
staining protocols.
protocols. Basophils
Basophils
rare
rare in
in the
the circulation,
circulation, but
but are
are potent
potent respond-
respond- Phagosomes
inophils, basophils are thought to play a role Phagosomes

Phagocytes
Dendritic
inophils, basophils are thought to play a role
nse
nse toto parasites,
parasites, particularly
particularly helminths (para-
helminths Cells
(para- Processes
Processes
When they bind circulating antibody/antigen Cells, Organs, and Microenvironments of the Im
When they bind circulating antibody/antigen Lysosome
asophils release the contents of their granules. Lysosome
asophils release the contents of their granules.
ne of the best known compounds in basophilic
ne of the best known compounds in basophilic (d)Neutrophil
(a) Megakaryocyte (c)
reases blood vessel permeability and smooth (d) Megakaryocyte
reases blood vessel permeability and smooth Multilobed
ty, and allows immune cells access to a site of Megakaryocyte
ty, and allows immune cells access to a site of Megakaryocyte
nucleus
sophils also release cytokines that can recruit
sophils
ne cells,also release eosinophils
including cytokines that
andcan recruit
lympho-
Neutrophils Granules
ne cells, including eosinophils and lympho-
s where parasitic worm infection is less prev- Platelets
s where Platelets
ines are parasitic worm infection
best appreciated is less
as a cause prev-
of allergy Phagosome
ines are best appreciated as a cause of allergy
s (see Figure 2-4d) also play a role in combat- FIGURE 2-5 Examples of monocytes, macrophages,
FIGURE
(b) Eosinophil2-5 Examples of monocytes, macrophages,
 Macrophages are found in many tissues and
some tissue-resident macrophages have different names
 Monocytes and Macrophages
Blood Monocyte

• Macrophages derive from monocytes in the blood (except for self-

renewing types, previous slide)

• When monocytes reach the tissue, they differentiate into macrophages

with lots of protrusions (pseudopodia) to seek out pathogens

• Ingest and kill pathogens (“Macro” = Big, “Phage” = Devour/Eat)

• Remove aging, dying, and dead cells (recognition of DAMPs)

Macrophage

• Produce inflammatory cytokines

• Antigen-Presenting Cell (APC) for activated T cells

• Longer life than neutrophils, sustain inflammatory response for several

days
Dendritic cells (DCs)

• DCs are resident in mucosal and cutaneous tissues to monitor major

routes of pathogen entry

• Distinctive feature of DCs: Huge extensions or “dendrites” for

scanning surroundings for pathogens

Major roles of DCs:

• PRR-mediated uptake of pathogen
• Degrade pathogens
• Travel via lymphatics to local lymph nodes for antigen presentation
to naïve T cells

DCs are main ‘antigen-presenting’ cell (APC) for naïve T cells

à key to ‘priming’ adaptive immunity

Innate and adaptive immunity work together to clear infections
Neutrophils
The first leukocytes recruited by
inflammation

• High number in blood (50-70% of circulating leukocytes)

• Also called polymorphonuclear leukocytes (PMNs) Neutrophil ingesting
S. aureus
• Main role: Get to site of infection rapidly, enter tissues,
use phagocytic receptors to ingest and kill extracellular
microorganisms
• Cytoplasm is full of granules that contain toxic products
to kill pathogens
• After taking up microorganisms and releasing their
granules, the neutrophil will die

Neutrophil ingesting S. aureus

NIAID / CC BY (https://creativecommons.org/licenses/by/2.0)
Neutrophil migration to site of injury

Courtesy of Dr. Paul

Kubes, University of
Calgary
Phagocytosis and intracellular killing of pathogens
Neutrophils can kill microbes using different mechanisms

Neutrophils phagocytose bacteria and kill them

within the phagosome:

• Antimicrobial peptides
• Lysozyme (an enzyme that degrades bacterial
cell walls)
• ROS (through the respiratory burst)

Netosis:
Formation of neutrophil extracellular traps
(NETS) that serve to trap bacteria, facilitating
their killing.
“Dying for a cause”: Neutrophils die and trap pathogens in NETs

DNA Bacteria

• NETs = neutrophil extracellular traps,

chromatin structures loaded with
antimicrobial proteins
• As neutrophil dies it extrudes its DNA
• DNA is very sticky and traps bacteria, killed by
the antimicrobial proteins
• Macrophages clean up the area

Dying neutrophil
Formation of Reactive Oxygen Species (ROS) in Phagocytes
is mediated by the enzyme complex NADPH oxidase
(Respiratory burst)

ROS are powerful oxidizing

factors that kill pathogens
 Defects in killing bacteria: Chronic granulomatous disease (CGD)

• CGD results from mutations affecting NADPH oxidase (the multi-

subunit enzyme that generates ROS).
• Most common mutation is X-linked thus disease mainly affects
males.
• Neutrophils and macrophages from CGD patients cannot
undergo the respiratory burst to generate ROS
• Microbes cannot be killed and patients have recurrent infections
• ‘Granulomas’ are formed (in lungs, lymph nodes, skin, bowel)
• Cyst-like formations of immune cells surrounding
undigested microbes
• Microbial clearance has failed so the body ‘walls off’ the
microbe to prevent spread
 he blood into the tissue spaces. They are most Phagosome
Phagosome
ant in the small intestines, where their role
ant in the small intestines, where their role is still is still
nvestigated.
nvestigated. In In areas
areas where
where parasites
Phagocytes:
parasites are
are less
less ofof aa (b)
A Summary
Macrophage
(b) Macrophage
problem, eosinophils are better appreciated
problem, eosinophils are better appreciated as con- as con-
ors Pseudopodia
ors to asthma
asthma and
toMacrophages and allergy
allergy symptoms.
symptoms. LikeLike neutro-
neutro- Pseudopodia
osinophils
osinophils may
may also
Main functions: secrete
secrete cytokines
also phagocytosis
cytokines that
that regulate
and killing regulate
of Phagosomes
Phagosomes
T
T lymphocytes, thereby
pathogens, cytokine
lymphocytes, influencing
therebyproduction, the
the adaptive
influencingantigen adaptive Lysosome
Lysosome
ne response.
presentation
ne response. Phagolysosome
Phagolysosome
ophi ls are nonphagocytic granulocytes (see
ophils are nonphagocytic granulocytes (see
2-4c) that contain large basophilic granules that
2-4c) that contain large basophilic granules that (c) Dendritic cell
(c) Dendritic cell
lue in standard H&E staining protocols. Basophils
lue in standard H&E staining protocols. Basophils
tivelyDendritic
rare in the circulation, but are potent respond-
Cells
tively rare in the circulation, but are potent respond- Phagosomes
ke eosinophils,
Main basophils
functions: are thought
phagocytosis and to play a role
cytokine Phagosomes
ke eosinophils,
response to
basophils
parasites,
are thought
particularly
to play (para-
helminths
a role
responseproduction,
to best antigen-presenting
parasites, particularly cell, bridges
helminths (para- Cells, Organs, and Microenvironments
Processes of
Processes
orms).innate
Whenand they bind
adaptive circulating antibody/antigen
immunity antibody/antigen
orms). When they bind circulating
exes basophils release the contents of their granules. Lysosome
Lysosome
exes basophils release the contents of their
ine, one of the best known compounds in basophilic granules.
(a) Neutrophil
ine, one of the best known compounds
es, increases blood vessel permeability and smooth in basophilic (d) Megakaryocyte
(d) Megakaryocyte Multilobed
es, increases
activity, blood vessel permeability and
and allows immune cells access to a site of smooth Megakaryocyte
Neutrophils nucleus
on.activity, and allows
Basophils immune
also release cells access
cytokines to arecruit
that can site of Megakaryocyte
Also classified as Granulocytes
on.
mmuneBasophils also release eosinophils
cytokines that can recruit Granules
Maincells, including
functions: and
phagocytosis and killing lympho-
of
mmune
n areas cells, parasitic
where includingworm eosinophils and lympho- Platelets
pathogens, NET formationinfection is less prev- Phagosome
Platelets
n areas where
histamines are parasitic worm infection
best appreciated is less
as a cause prev-
of allergy
histamines are best appreciated as a cause of allergy
oms.
 Three Types of Granulocytes

Neutrophil Eosinophil Basophil

Cytoplasmic granules stain differently in the three different cell types

with the hematoxylin-eosin stain commonly used in blood smears
Some pathogens cannot be handled by phagocytosis as a defense

Phagocytosis is suited for microbes that are

small and extracellular

Copyright © 2006 Elsevier Inc.

What about a pathogen that is extracellular,

but very large; much larger than a phagocytic
Photo credit: E.M.Unit, Royal
Free Hospital, School of
cell? (e.g., parasitic worms)
Medicine/Wellcome Trust
Photo Library
Degranulating Cells
Mast cells
• Contain granules but historically not classified as
granulocytes because not found in the blood
• Reside in tissues
• Involved in parasitic infections and allergy
• Release histamine and other mediators

Basophils
• Least common of the innate immune leukocytes
• Are granulocytes like neutrophils, with many toxic
substances contained in granules
• Produce histamine
• Involved in defense against parasites
• Recruited from blood during allergic responses

Eosinophils
• Also granulocytes
• Involved in parasitic infections and allergy
• Granules contain toxic cationic proteins
Degranulation products are responsible for
immediate allergy symptoms

• Designed to eject parasites from the body, but in response to

harmless entities (e.g., pollen) can cause symptoms of allergy

• Immediate effects due to release of histamine and other

preformed inflammatory mediators (especially by mast cells)

• Cause vasodilation, mucus production, itching, gut motility

Pixaby, PublicDomainPictures
 FceRI-mediated degranulation of mast cells

Degranulation of mast cells takes place within seconds of IgE-mediated antigen recognition
because mast cells are pre-loaded with IgE attached to their Fc receptors
Natural Killer Cells (NK)

• NK cells are large, granular, cytotoxic lymphocytes (but they

are not T or B lymphocytes)

• First line of defense against intracellular pathogens

• Virally infected cells
• Intracellular bacteria/protozoa
• Cancer cells

• Release of cytolytic granules causes targeted cell destruction