Innate and Adaptive Immunity

Dr Brian Brestovac

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 Immune System
• Innate:
– Non specific defences which do not require
previous exposure to foreign agent

• Adaptive:
– Response based on specific recognition of invader
(foreign agent)
• Antibody mediated (humoral)
• Cell mediated
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 Human Immune Sys tem

Innate A daptive
Innate and Adaptive braches talk

Humoral Cell mediated

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 Introduction to Innate Immunity
30-Jul-15 PJC 7
Recognition of pathogens by receptors/molecules of innate immune system
Are not present as a response to specific stimulating event
Present from birth (Genes code for proteins used in Innate system)
Immediate - No memory
It recognises surface molecules on microbes that are common to many pathogens (Activated by a
limited set of molecular structures)
It exists in all living multi-cellular animals (Very old system – evolved long ago)
Sometimes referred to as:
Inborn
natural

• Relatively non-specific
• The innate immune system provides an initial discrimination between self and non-self
- The goal is to inhibit the spread of invaders – “shoot first and ask questions later”-

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 Functions of the Innate System
1. Initial defence against microbes
– Prevents spread, eliminates, control

2. Eliminates damaged cells and initiates repair

3. Stimulates adaptive immune response

– Provides danger signals
– Influence the type of adaptive response
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1. Surface Barriers
• Physical barrier – skin & mucosal
• Mucous membrane barriers trap microbes carried to external openings by ciliated epithelial cells
- Cough reflex stops entry into the RT
- GIT: hostile environment – acidic pH, proteolytic enzymes etc

2. Internal defence
• Destruction of the invader/foreign entity by specialist cells
- Macrophages
- Neutrophils
- Dendritic cells
• NK cells

• Inflammation
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Anti- bacterial peptides
Evolved early
Small 6 – 59 amino acids, positively charged (cationic) with anti-microbial activity
Defensins (humans)
Kill a wide range of bacteria within minutes
Some fungi and enveloped viruses
Secreted by neutrophils, epithelial cells and paneth cell (intestine)
Disrupt microbe membranes
binds and forms a pore – lysis
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 Inflammation
• Important part of innate immune response
• Many processes start with inflammation
– Triggered by tissue/cellular injury
• Infection, physical trauma, chemical, heat
– Attempt to prevent spread of agent
– Disposal of debris and pathogens
– Tissue repair
– Alerts adaptive immune system
• Over-response can do damage
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 Response
Injury!!
Cells damaged – die
(plus local mast cells, Macrophages)

Release Inflammatory Mediators

Acute inflammation
Damaging stimulus removed

Cells regenerate Cells do not re-grow

Restoration of normal Healing by repair
structure and function Scar formation
(fibrous repair – loss of 10
function)
 Cardinal signs of Inflammation

1. Redness
2. Heat
3. Swelling
4. Pain
5. Loss of function

Pus – masses of WBC mostly neutrophils (first

Red, swollen knee with pus (WBC) oozing
WBC type to go to a site of damage)
out – looks painful, hot and probably isn’t
Notice the microbes phagocytosed by the 11
very functional
neutrophils
 Inflammatory mediators
• Cytokines
– Proteins released by cells that affect other cells
• interleukins, interferons (IL-1, IL-6, TNFα, IL-8, INFƔ)

• Chemokines
– Proteins released by cells that attract other cells to area

• Acute-phase proteins
– Plasma proteins that increase in concentration due to
inflammation within minutes
• C-reactive protein binds microbes and activates complement
• Kinins act locally to induce vasodilation

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 Phagocytosis
• Microbes are killed and removed
– Phagocytic cell recognises surface of
microbe, adheres and takes in microbe
– Held within a phagosome which fuses
with a lysosome to form a
phagolysosome
– Toxic substances kill and degrade
microbe
• Nitric oxide
• Superoxide anions (O2-)
• Hydrogen peroxide (H2O2)

– Exocytosis to remove microbe debris

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 Natural killer (NK) cells
• NK cells eliminate both virally infected and
cancer cells
– NK cells recognize reduced MHC class I
molecules via activating and inhibitory
receptors
– Antibody Dependent Cell Mediated Cytotoxicity
(ADCC)
• NK cells can attach to antibodies (via Fc receptor)
on surface of cells

– Direct cytotoxicity by release of perforin and

granzymes
 NK cells
“The missing self hypothesis”

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Stimulation of the Innate System
PAMPs
– Pathogen-associated molecular patterns
– Different structures on different pathogens
that are recognised by cells of the innate
system
– These structures often are essential to
pathogen
– Stimulates a response
– Limited in number
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Stimulation of the Innate System
DAMPs
– Damage-associated molecular patterns
– Molecules released from damaged cells
– Endogenous
– Released in response to trauma, toxin, burns,
infection etc
– Healthy cells can release alarmins in
response to infection which enhance innate
response
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 PAMPs DAMPs

Nucleic acid ssRNA, Viruses Stress proteins HSPs

dsRNA,
CpG
Proteins Pilin, Bacteria
Flagelin
Cell wall lipids LPS GNB Crystals Monosodium urate

Lipoteichoic GPB
acid
Carbohydrates Mannan, Fungi Nuclear High-mobility
Glucans proteins group box 1
(HMGB1)

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Receptors of the Innate System
• Pattern Recognition Receptors (PRR)

– Cellular Phagocytes (macrophages &

neutrophils, DC, endothelial cells)
• When bind to PAMPs/DAMPs activate signal
transduction pathways
• Promote inflammation and antimicrobial activity
– Soluble molecules
• In blood and extracellular fluid
• Enhance phagocytosis
• Active extracellular killing 19
 Cellular PRR
PRR Location Example PAMP/DAMP
Toll-like receptors (TLR) Membranes of DC, TLR 1-11 Microbial molecules
Phagocytes, B cells, (LPS, viral nucleic acid etc)
endothelial cells

NOD-like receptors (NLR) Cytosol DOD 1/2 Bacterial cell walls, crystals
(phagocytes, other
cells)

RIG-like receptors (RLR) Cytosol RIG-1, MDA-5 Viral RNA

(phagocytes, other
cells)

Cytosolic DNA sensors Cytosol of many cell AIM2, STING- Bacterial & viral DNA
(CDS) types associated CDS’s

C-type lectin –like receptors Membranes of Mannose receptor, Microbial surface

(CLR) phagocytes Dectin Carbohydrates
Gylcans (Fungi)

Scavenger receptors Membranes of CD36 Microbial diacylglycerides

phagocytes

N-Formyl met-leu-phe Membranes of FPR Peptides

receptors phagocytes
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 Soluble PRR
PRR Location Example PAMP/DAMP

Pentraxins Plasma C-reactive protein Microbial

phosphylchlorine &
phatidylethanolmine

Collectins Plasma Mannose binding Carbohydrates with

lectin terminal mannose
Microbial structures
Aveoli Surfactant proteins

Ficolins Plasma Ficolin Cell wall components

of Gram positive
bacteria

Complement Plasma C1 – C9 Microbial surfaces

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 TLR
Bacterial Bacterial
Flagellin lipoproteins ?
LPS
Bacterial
peptidglycan Profilin

Bacterial TLR6
lipoproteins TLR5
TLR4 TLR10

TLR2
TLR11
Endosome
TLR1 TLR7

ssRNA

TLR3
CpG
dsRNA DNA
TLR9
Nucleus
ssRNA

TLR8

TNF, IL1, IL6

Acute inflammation
Adaptive Immunity IFN α/β
Antivirus 22
 NOD-like receptors (NLR)
Nucleotide binding Oligomerization –like receptors

• Specific for intracellular pathogens

– In the cytosol of cells

– Activated by peptidoglygan, RNA, toxins,

flagellin

– Release pro-inflammatory cytokines

• IL-1, IL-6, TNFα, IL-8

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 Interferons (IFN)
– Broad spectrum anti – viral agents
– Three groups IFNα (14 types), IFNβ and IFNγ
• IFNα & IFNβ produced in response to viral infection
• IFNγ - inflammatory (Macrophages, B cells, TH1)

– Binds specific receptors of neighbouring cells

• Neighbouring cell produces Protein Kinase R (PKR)
which blocks virus entry
• Limits spread of viral infections
• Stops production of protein in cell (inhibits virus
production)
• Stimulates endonuclease production which degrades
viral mRNA
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 Antigen Presenting Cells
(APC)
• Cells that phagocytise infectious invader (antigen)
• Migrate to lymph node
• Digest and present bits of antigen to Thelper cells (via MHC II)
• Professional antigen presenting cells that move around
detecting foreign antigens
– Dendritic cells
• (the most efficient antigen presenting cell)

– Macrophages

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Dendritic cell
 Adaptive Immunity
• Innate immunity does not improve or amplify
with repeat exposure
• Once overcome innate immunity ineffective
• Adaptive immunity fight invaders once innate
system breached
• More specific than innate system
• Has memory
• Mediated by lymphocytes

– DC and Macrophages deliver message to adaptive

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system from innate system
 Adaptive Immune System
• Divided into two, although there is overlap
and communication
1. Humoral Immunity
– Antibodies mediated
– Action against extracellular invaders
– Produced by B lymphocytes
2. Cell Medicated Immunity (CMI)
– Lymphocytes directly / indirectly kill infected cells
– Act against intracellular invaders (also cancer)
– Cytotoxic T cells (Tc)

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 T cell receptors and
antibodies
• These proteins have a massive repertoire!
– Need to deal with a vast number of antigen!
• Very specific!!
• Not enough genes to produce this diversity
• Somatic cell recombination
– Multiple gene segments combined and
recombined to form variations in protein
binding of antigen
– Only T cell receptors and antibodies
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 Humoral Immunity
• Antibody mediated immunity
• Antibodies are specialised proteins that recognise
and attach to antigen
• Recognise antigen on a foreign invader
• Action of Antibodies:
– Activate complement
– Trigger phagocytosis (opsinisation)
– Neutralise viruses and toxins (so that they can not enter
cells)
– Aggultination (aggregation)

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 Classes of Antibodies (Isotypes)
• IgG
– Monomer, most abundant in serum
– largest amount, long term immunity, can cross placenta
• IgA
– Monomer in serum, dimer in saliva and secretions, most abundant
total
– saliva and mucosal surfaces, tear, nasal fluids, milk
• IgM
– Pentamer, primary response
– first Ab to appear during an infection
• IgE
– Monomer, stem binds mast cells, basophils and eosinophils causing
release if granules
– anti-parasite, allergy
• IgD
– Monomer, attached to B cells, antigen receptor for activation 30
– largest Ab - Ag receptor on B cells
Optimisation

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 Primary and Secondary Immune
Responses
• Primary response
– Primary response has a lag of several days while B cells proliferate (time
for invader to do harm)
– Antibody levels peaks and falls as antigen is removed
– The type of antibody is predominantly IgM (Immunoglobulin M)
• Secondary response
– Second exposure to same antigen
– The response is much quicker and larger with a rapid rise in antibody
levels and lasts longer
– Memory cells require fewer cycles to become plasma cells (shorter
reaction time)
– The antibody type is predominantly IgG (Immunoglobulin G)

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 Clonal Selection
• Antigen can only bind to an antibody specific for it
• Antigen does the selecting
• Specific antigen binds to the variable region of B cell surface
antibody
• The B cell (with the specific surface antibody) is stimulated
• Clone is selected
• Other antibodies on other B cells don’t bind (poor match)

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Primary and Secondary Immune Responses

Without T cell help B cells only produce IgM in low

quantities (T-independent).
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 Cell Mediated Immunity
• Humoral immunity only effective against extracellular antigen

• Viruses and mycobacteria are intracellular infections (out of

reach of humoral response)

• Cancer cells also need to be removed

• The mechanism for protection against these attacks is the Cell

Mediated Immune (CMI) response

• The populations of T cells involved:

– Cytotoxic T cells (TC)
• Require Thelp (Th)
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 Cytotoxic T cells (TC)
• Wide range of surface receptors against antigen - T cell receptors
(TCR)

• Not immunoglobulin as in B cells

• Each TC is specific for one receptor

• Infected cells express the antigen (of the invader) on their surface
by MHC 1

• TC via T cell receptor binds and destroys cells expressing the

specific antigen with the MHC 1

• Kill by release of toxic molecules

– effect membrane causing lysis,
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– initiate apoptosis
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 Lack of
NK MHC1 MHC 1

Virus
infected
Tc

Y
cells
NK

Antigen presented
Antibody by MHC 1

Adaptive (specific killing)

Tc Cells kill by detecting Ag presented by MHC 1
NK cells killing by detecting antibody on cell (ADCC)

Innate (non-specific killing)

NK cells by detecting lack of MHC 1 (missing self hypothesis)
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Adaptive Immune Response

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 Feedback study questions
1. List and describe the features of the innate immune system.
2. List and describe the features of the adaptive immune system.
3. How is the innate system stimulated and why does the innate system lack
specificity?
4. List the cardinal signs of inflammation.
5. In an inflammatory response what do cytokines, chemokines do?
6. How is the adaptive system stimulated and what makes it so specific?
7. If Toll-like receptor 3 (TLR3) is stimulated what is the outcome? What stimulates
TLR3?
8. Which TLRs are stimulated by bacterial proteins?
9. Where are the NOD-like receptors located?
10. Describe the two mechanisms by which NK cells are activated.
11. What do Dendritic cells and macrophages do? Are they part of the innate or
adaptive immune systems?
12. What are the down stream effects of an antibody binding to an antigen?
13. List the classes of antibodies and there function.
14. What are primary and secondary immune responses?
15. How do cytotoxic T cells (Tc) recognise cells for destruction?
16. Describe the immune steps that happen when a person is infected with agent.
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