Innate Immunity 1

Lecturer: Dr Bernadette Saunders

Senior Lecturer
School of Life Sciences
Email: Bernadette.Saunders@uts.edu.au

Copyright 2012 Hagop Kaneboughazian

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 Learning Objectives

After this lecture you should understand,

What are surface barriers and their immunological roles.

The characteristics of the innate immune system.
How the innate immune system responds to infection.
Internal defences, their characteristics and functions:
Phagocytosis
Innate immune cells
The complement system

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Innate Immunity: An evolutionary perspective

All eukaryotes have defence

mechanisms against
microbial & viral pathogens

Vertebrates have evolved

two types of immune
systems:

Innate and adaptive/acquired

immunity
Image by Olsen, V. Beth Kuser. 1997. Evolutionary Tree.
Retrieved in 2010 from V. Beth Kuser Olsens Homepage.
Cecil College: Maryland

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 Immune Response to Infection
Physical Barriers
Skin & Mucosal Surfaces (eg gut and
Invasion respiratory tract)
& infection

Innate immune Response Cells- Neutrophils, monocytes/

macrophages, NK cells,
cytokines/chemokines ie TNF, IL-6

Inflammation
Cellular and Humoral Responses
Adaptive immune Response Antibodies, cytokines, T helper cells,
cytotoxic T cells

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 Characteristics of Innate immunity

Ancient: present in all eukaryotes

Immediate/rapid response to pathogen
Recognition of molecular patterns common across
pathogens
Low specificity
Lack of memory
Directs the adaptive immune system

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 First line of defence: surface barriers
Skin
Heavily keratinised: physical barrier
resists colonisation
Weakly acidic (pH 3-5) which inhibits
bacteria/fungi growth
Sebum secreted by the sebaceous gland
contains antimicrobial substances
Mostly impenetrable unless broken

http://www.entwellbeing.com.au

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 First line of defence: surface barriers
Mucosal Membranes

Line all body cavities that open to the exterior

(digestive, reproductive, respiratory and urinary
tracts)
Secretion of HCl and enzymes into the stomach
that destroy pathogens
Enzymes in saliva (e.g. lysozyme) that break
down microbes

Mucociliary clearance video:

https://www.youtube.com/watch?v
=HMB6flEaZwI

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 First line of defence: surface barriers

Staphylococcus
Mucosal membranes
Production of mucus that
traps microbes (also Mucous
contains numerous anti-
INTERNETMEDICINE.COM
microbial molecules)

Modifications such as cilia

which trap and sweep away
microbes from the lower
respiratory tract

www.webmd.com
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 First line of defence - Epithelial cells
Functions within seconds of contacting a
pathogen
A mechanical, selectively permeable
barrier between the outside and inside
Produce natural antibiotics - defensins
(released from skin cells and neutrophils)
Outside
May possess motile cilia
Rapidly renewable
Produce cytokines - alter the behaviour of
other cells
Inside
Produce chemokines - attract other cells
Primary role is to block the entry May produce mucins
of microorganisms
Transport antibodies from inside to
outside
 Antimicrobial peptides (AMPs)

Two families of AMPs: cathelicidins and defensins

Cationic (positive/basic charged) short polypeptides found on

epithelial surfaces

AMPs bind to bacterial membrane

Oligomerisation of AMPs creates a pore in the bacterial

membrane = lysis of bacteria

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 Other types antimicrobials found in the
epithelium

Immunoglobulins especially IgA

Part of the adaptive immune system
Bind to bacteria (opsonisation) and enhance phagocytosis

Surfactants
Bind carbohydrates expressed by bacteria and enhance phagocytosis

Normal flora found in the epithelium - competition with

pathogenic microbes

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 Immunology in the skin

https://www.youtube.com/watch?v=_VhcZTGv0CU

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 Summary of antimicrobial barriers

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 Pathogen breaches epithelium

Humoral response Cellular response

Complement Phagocytes
C Reactive protein NK cells
Granulocytes
Pathogen persists

Antigen presentation and

adaptive immune response

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 Pathogen damages a physical barrier
- initiate innate immune response
Pathogen -damages the epithelium
breaks through the epithelial barrier

Epithelial cells activated upon

bacterial contact

Activated epithelial cells produce

chemokines and cytokines

Attract leukocytes to site of

infection
 Timing of innate immune response
Barriers - Epithelial activationComplement
- - Cytokines/chemokines -
Seconds Minutes Minutes Minutes to days

Inflammation
Neutrophils - Monocytes/macrophages - NK cells -
Hours Hours to weeks Hours to days

Produce cytokines
-IFN-
 Mediators of the immune system

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 Cells of the innate immune system

Phagocytes
Neutrophils
Macrophages
Dendritic cells

NK cells

Granulocytes
Neutrophils
Eosinophils
Basophils

Blausen.com staff. "Blausen gallery 2014".

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 What are phagocytes?

Phagocytosis (Ancient Greek)

Phagein = Devour,
Cytos = Cell

Any cell that is capable of ingesting a foreign particle

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 Types of phagocytes

Granulocytes (Mainly Neutrophils)

Macrophages

Immature Dendritic cells

http://garlandscience.com/garlandscience_resources/resource_detail.jsf?landing=student&resource
_id=9780815342434_CH03_QTM01

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 What are granulocytes?
Type of leukocyte characterised by
granules in their cytoplasm

Neutrophils, basophils and

eosinophils

Granules are intracellular vesicles

that contain a variety of
antimicrobial substances released
upon activation
Defensins
Proteolytic enzmes
www.medsci.uu.se
Cytokines

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 Neutrophils

Differentiate from multi-potential stem cells in BM

Released into blood in a mature form containing bactericidal products
Up to 1011 neutrophils are produced every day but are short-lived
Constitute 30-70% of WBCs in peripheral blood
Contain lysosomal granules containing lysozyme, acid hydrolyses, proteases,
peroxidase, etc.
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 Neutrophils
4 to 10 million per ml of blood
Infection cytokines ->stimulate bone marrow to produce up
to 20 million neutrophils per ml of blood.
Functions
Invade site of infection rapidly
Ingest bacteria
Produce antimicrobial products including ROI and RNI
(reactive oxygen intermediates and reactive nitrogen
intermediates, toxic to bacteria)
2000+ granules per cell-includes Peroxidase, Defensins
Neutrophil Extracellular Traps (NETs)
NETS are formed by activated neutrophils.
Contain a DNA backbone with antimicrobial peptides and enzymes
Involved in defence against a number of extracellular organisms
including pneumococcal pneumonia and streptococcal necrotizing
fasciitis.

Nature Reviews Immunology 2011 doi:10.1038/nri3024

 What happens when a macrophage encounters a
microbe? e.g. bacteria

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 Macrophages

In blood monocytes can mature into macrophages

In tissues they are specialized cells: Kupffer cells in liver
Differentiate from BM stem cells, released as monocytes: both phagocytic and
bactericidal
Constitute 3-7% of WBCs in peripheral blood
Emigrate to tissues, differentiate into macrophages
Highly phagocytic and contain numerous antibacterial responses
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 Phagocytosis
Phagocytic cells- macrophages,
neutrophils, dendritic cells.

Phagocytosis - active process -bacteria

and foreign bodies are engulfed and
internalised into a membrane-enclosed
vesicle known as a phagosome.
The phagosome becomes acidified,
fuses with lyzosomes, and kills the
microbe.

Internalized microorganisms and lysed

products are recognized by PAMPs
(Pathogen-associated molecular pattern
receptors). (more about these next
lecture)
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 Phagocytosis

Also complement receptors found on phagocytes

enhances phagocytosis of complement coated pathogen
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 Macrophages

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 Bacterial clearance by phagocytosis

http://garlandscience.com/garlandscience_resou
rces/resource_detail.jsf?landing=student&resou
rce_id=9780815342434_CH11_QTM01

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 Antibacterial Mechanisms in Macrophages
IDO P2X7
Reactive
Oxygen
Intermediates Vit D3

Phagosome
Acidification/ Antimicrobial
Phago- Peptides
lysosome
fusion
Autophagy
Nutrient
Deprivation Reactive
eg iron Nitrogen
Apoptosis
Intermediates
 Dendritic Cells (DCs)

There are different types of DCs present in different locations in

the body
Main function is to take up and process antigenic material and
present it to the immune system (bridge between the innate and
adaptive immune system)
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 Dendritic Cells:
heterogenous (there are different
types of DCs,... in different places)
can be derived from monocytes
(monocyte-derived DCs)
they are endogenous within the
skin (Langerhands cells & dermal
DCs)
are endogenous within all tissues
(interstitial DCs)
in blood they are massive
producers of IFN (plasmacytoid
DCs)

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 Dendritic cells

DC (green) interacts with a T cell (light purple) Bashyam, The Journal of

Experimental Medicine
2007
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 Dendritic cells act as a bridge between
innate and adaptive immunity
Dendritic cells endocytose/phagocytose pathogen (antigen)
Digest pathogen into small peptides
Peptides are bound to MHC and then transported to the cell surface for
presentation to antigen-specific T cells

Copyright UTS Abbas & Lichtman, Elsevier 38

 Dendritic Cells
Immature DC are highly phagocytic in tissue-
express low levels of Class II and co-stimulatory
molecules CD80 & CD86, CD40
Upon phagocytosis they:-
-up regulate CD80/86 & Class II expression and
CCR7 ( important in homing to the lymph-node)
-Migrate to draining lymph nodes
-Present Ag to T cells
-Produce cytokines including IL-12, IL-23, TNF, IL-1,
IL-6 and IL-10
Crucial for inducing the activation and proliferation of naive
T cells and are involved in T-cell differentiation and
polarization.
 Basophils

Contain large basophilic granules containing histamine, heparin,

proteolytic enzymes
Least common granulocyte, they constitute 0.1-0.3% of WBCs in
peripheral blood
Important role in certain inflammatory reactions
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 Eosinophils

Contain eosinophilic granules containing histamine, peroxidase,

lipases, etc
They constitute 1-5% of WBCs in peripheral blood
Important role in parasite infections, allergy and asthma

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 Humoral Immunity
Soluble factors (non-cellular) that mediate immunity
generally proteins

Three broad types:

Antibodies
Complement system
C-Reactive Protein (CRP)

Antibodies are part of the adaptive immune system (although

can have innate properties)

Complement and CRP are part of the innate immune system

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 Antibodies eliminate extracellular
microbes and toxins
Antibodies cannot enter host cells

Extracellular microbes, that colonise and replicate in

extracellular tissue spaces, fluids etc. are vulnerable to
antibodies

Antibodies use various mechanisms to eliminate extracellular

microbes
Effector functions of antibodies
Basic Immunology Functions & Disorders of the Immune System 2nd Edition 2004
By AK Abbas & AH Lichtman, Saunders, Elsevier. Figure 8-1A
 Complement System
Group of >30 soluble proteins (mostly enzymes) found in
serum in inactive form (zymogens)

Frequently referred to as a cascade sequential activation of

complement proteins by cleavage
A deficiency in one complement protein can affect the whole system

Three pathways:
Classical
Lectin
Alternative

Results in microbial killing, opsonisation and inflammation

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 Initiation
Amplification
Termination

Dunkelberger & Song, Cell Research, 2010

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 Three pathways of
complement activation
Classical pathway activated by IgM & IgG bound to
microbes or in Ag-Ab complexes

Alternative pathway, triggered by C3b and factor B bound to

microbes (innate pattern recognition)

Lectin pathway, triggered by mannose binding protein

(lectin) binding multiple mannose residues in some microbe
cell walls (innate pattern recognition)
 Complement System

Abbas & Lichtman, Elsevier

Pathways converge at the generation of C3 convertase

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 Effector functions of the complement system
Direct lysis of pathogens via the Membrane Attack Complex
(MAC)
Complement components form a complex, exposure of hydrophobic
phase allows insertion to microbial membrane. C9 binds to complex
and polymerizes forming a pore

Dunkelberger & Song, Cell Research, 2010

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 Effector functions of the complement system
Enhancement of phagocytosis (opsonisation). Receptors on the
surface of phagocytic cells recognise complement components attached to
the microbial surface or to antibodies (which coating the microbe).

Dunkelberger & Song, Cell Research, 2010

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 Effector functions of the complement system
Recruitment and activation of immune cells
Interaction of complement fragments (anaphylatoxins C3a and C5a) with
receptors on endothelial cells and mast cells produce local inflammatory
responses.
Phagocytic cells are recruited

Dunkelberger & Song, Cell Research, 2010

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Effector functions of activated
complement components
 Learning Outcomes
Innate Immunity
Evolutionarily conserved in eukaryotes
Immediate with low specificity
Barriers: Skin, mucosal membranes, normal flora (microbiota)
Internal defences: antimicrobial peptides, complement,
immunoglobulins and phagocytosis
Complement: classical, lectin and alternative pathways. Effector
functions: inflammation, opsonisation and Membrane Attack
Complex
Phagocytes: internalize pathogens for their clearance and processing
into antigenic peptides; include Neutrophils, Macrophages and
Dendritic Cells
Granulocytes: contain antimicrobial granules; include neutrophils,
eosinophils and basophils
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Revision Questions
 Revision
Question 1.
Which of the following innate immune response
statements is false?
a. Surface receptors of innate immune cells recognise
pathogen associated molecular patterns.
b. Results in the production of pathogen-specific
antibodies and long term memory.
c. Innate immunity enables rapid/immediate responses
to invading pathogens.
d. Includes internal defences such as phagocytosis of
invading pathogens by innate immune cells.

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 Revision

Question 2.
Which of the following statements is true for antimicrobial
peptides (AMP)?
a. Enhance phagocytosis.
b. Preferentially bind eukaryotic membranes.
c. Perforate bacterial membranes to induce lysis.
d. Enhance phagocytosis and preferentially bind eukaryotic
membranes.

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 Revision

Question 3.
Which of the following statements is specific to gut chemical
barriers?
a. Specialized enzymes such as pepsin.
b. Antimicrobial peptides.
c. Lysozymes.
d. None of the above.

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 Revision

Question 4.
Which complement pathway becomes activated by
spontaneous hydrolysis of C3?
a. Classical
b. Alternative
c. Lectin
d. Both options a & b are correct

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 Revision

Question 5.
Complete the following sentence. The cleavage of C3 and C5
results in the production of C3a and C5a fragments which
are.......
a. fundamental for the establishment of the membrane attack
complex (MAC).
b. inactive molecules.
c. opsonizing agents.
d. anaphylatoxins.

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 Revision
Question 6.
Which of the following statements is true for
Neutrophils?
a. Emigrate to tissues and become resident neutrophils with
specialised functions.
b. Are the most abundant white blood cell (WBC) in
peripheral blood.
c. Are the most efficient antigen presenting immune cell
(APC).
d. Kill virally infected host cells by forming pores in their
extracellular membrane.

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 Revision

Question 7.
Which of the innate immune cells below are not phagocytic?
a. Lymphocytes
b. Neutrophils
c. Dendritic cells
d. Macrophages

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 Revision

Question 8.
Which of the following statements best describes the
classical complement pathway effector function?
a. Inflammation.
b. Pathogenic lysis.
c. Opsonisation.
d. All of the above.

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 Revision

Question 9.
Which complement component is involved in all
complement pathways?
a. C9
b. C3
c. C1q
d. C4

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 Revision

Question 10.
Which of the following options best describes an
opsonising agent?
a. Surfactants
b. Antibodies
c. C3b
d. All of the above

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 Revision

Question 11.
Which is the least abundant granulocyte in peripheral
blood?
a. Basophils.
b. Mast Cells.
c. Neutrophil.
d. Eosinophils.

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 Revision
Question 12.
Which innate immune cell efficiently kills antibody
coated parasites?
a. Macrophages
b. Eosinophils
c. Neutrophils
d. Basophils

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 Revision
Question 13.
What is the vesicle into which a bacteria is initially
engulfed by a macrophage
a. lysosome
b. endosome
c. phagosome
d. phagolysosome

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