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Innate Immunity
Learning Outcomes
- Infection occurs only after the organism or infectious agent has gained entry
into the body
o 1. List the ways in which an organism or infectious agent can gain entry
into the body?
Ingestion: eating/drinking (food poisoning)
Inhalation: respiratory pathogens
Lesion: wound, surgical lesion
Mammary glands (e.g. mastitis in cows)
Urinary tract
Ear, nose, eye, etc.
Insect bite
o 2. List the ways in which an organism or infectious agent can then spread
internally in an animal?
Skin is a really good barrier and there are other aspects of the
barrier system that are quite protective and good at stopping
infection infectious agents have ways of getting around this:
Virulence factors (e.g. to allow
for adhesion)
Produce toxins at site of infection
(e.g. enterotoxogenic E. coli)
Target particular cells (some
pathogens will target
macrophages or T cells to hide
from the immune response)
Form biofilms (surface covering,
no longer exposed to the
immune response)
Extension from original site
Target tissues
o E.g. sheep with cheesy
gland, the pathogens
target the lymph nodes
and then cause necrosis at
that site through the
production of toxins
Blood stream: if there is a lesion or wound that goes directly
into the vasculature, or particular pathogens like anthrax can
target the blood stream
- Barriers
- Innate immune cells
o Granulocytes, monocyte/macrophages, mast cells, NK cells, dendritic cells
- Mediators (soluble)
o Antimicrobial secreted substances, inflammatory mediators, complement,
cytokines/chemokines
- Actions
o Phagocytosis, direct cell killing, opsinisation, inflammation, cell migration
(chemotaxis getting other cells in to help), initiation of downstream
immune responses
- Ski
n : has
epithelia
(great
barrier),
has tight
Epithelial cells
- Barrier
o Tight junctions
- Antimicrobial peptides: prevent the
bacteria from getting a foothold
- Sense pathogens
- Activation
o Epithelial cells can be activated to
produce chemokines (interleukin A (a
cytokine that is chemotactic) tries
to attract other cells in)
- Image:
o (3) Produce toxins that kill epithelial
cells
o (4) Destroy the tight junctions
o (7) Some have adhesive molecules
that allow them to gain entry in to the cells
- Very important as they are the front line (ready to go, no time lag in the defence
they provide)
- Provide protection early in infection
- Anti-microbial proteins/peptides
o Produced by epithelial cells and phagocytes
o Direct and immediate no time lag in defence
o Detect early infection
o Include:
Lysozyme: digests and breaks specific bonds in bacterial cell walls
Defensins: ancient, evolutionarily conserved antimicrobial peptides
that disrupt bacteria, fungi cell membranes and some viral
envelopes
Cathelicidins: made as inactive propeptides and cleaved to form the
active peptide when needed (can be targeted to where they are
needed)
Histatins: produced in the oral cavity
- Having the right diversity in your microbiome (commensals within the gut) may
act to be protective in a lot of different disease conditions
o Interactions between the microbiome and innate immune system: if you
have had a course of antibiotics and then are exposed to a viral pathogen,
there can be lack of responsiveness of the system, has suppressed that
interaction as the commensals have been wiped out as well as the
bacterial pathogen, is dampening down the interaction between the
commensal bacteria and the innate immune system at the site of the gut
this can have systemic effects. Its tempering the immune response so
that you have a level of responsiveness of your cells that you can instantly
respond to other pathogens
- Leukocyte is overarching term for all types of white blood cells; is not limited to
just the innate or adaptive cells (more broad term), can then break down into
subtypes:
o Neutrophils: front line defence, are phagocytic, have bactericidal
mechanisms (they produce a lot of antimicrobial substances), they set the
scene for further aspects of the inflammatory response
o Lymphocytes: subset of broader categorisation of leukocytes, are involved
in adaptive immunity
Are the B and T cells that give the memory response
o NK cells: important front line defence, they are surveyors, keep a constant
watch to detect anything abnormal, particularly important for tumour
responses, also important in viral responses, look like lymphocytes but are
part of the innate immune response (not the adaptive)
o Monocytes: differentiate into macrophages (in tissue) monocytes are in
blood, phagocytic, bactericidal, important in cell recruitment
- Mast cell: present within tissues not the blood, part of the innate immune
response
- Dendritic cell: present within tissues not the blood, part of the innate immune
response
- The definition of innate vs. adaptive is based on the specificity of the receptor
o The monocytes dont have those specific B or T cell receptors, but they
show the B or T cells what they need to be doing how they are
connected to the adaptive immune response
Toll-like receptors
- 1. Phagocytosis
o One of the main processes of the innate immune
system for clearing/getting rid of foreign
agents/bacteria
o The three main cell types involved are:
neutrophils, macrophages and dendritic cells
Macrophages are called different things
depending on which tissue they are in but
are all similar in action:
Liver: Kupffer cells
Phagocytes
o Dendritic cells: link the innate and adaptive immune responses, tissue
resident cells, involved (chiefly) in antigen presentation, can also
phagocytose pathogens
Adaptive immune response
- Used intravital microscopy: can look at what is happening inside an animal in
real time
- Injured site is labelled with red dye
- Neutrophils are green: many cells migrate to the site of injury to repair the
damage
o Very fast response, this is why neutrophils are the first line of defence
Summary
- Phagocytosis
- Complement system
- Inflammation
- Efficiency of phagocytosis is increased by complement an antibody bind to
surface of organism making it more attractive to phagocytic cells
- 2. Complement
o Originally used to describe the activity of molecules in serum which could
complement the ability of antibody to lyse bacteria
o If there was uncoated bacteria, they would be cleared but at a slower rate
o Then is antibody specific to the bacteria was added to the mixture
o And if a combination of antibody and these plasma proteins was added,
then the rate of killing of the bacteria was enhanced (augmented)
o This augmented aspect: now getting an interaction between both the
innate and acquired immune response; the antibodies are from the
acquired/adaptive immune response and the complement is part of the
innate immune response
Complement
Complement cascade
Assembly and regulation of the MAC based on structures of C5b6 and sC5b9
- Looked at C5 and C5b to see these would fit together how the pore would be
formed
- It caused a conformational change when it broke down from C5 to C5b, if there
was no membrane for it to bind to, then it activates itself so it doesnt form a
pore
- If there are the right conditions, get formation of MAC
- Direct/effector pathway for destroying the bacteria, also have an indirect
pathway where its opsonised and then you get phagocytosis (two-pronged
attack)
Complement system
What is inflammation?
Agent Type
Physical Foreign object (e.g. splinter),
trauma, heat/cold, radiant energy
(UV and sunlight)
Infectious Microbes (bacteria, viruses, fungi),
protozoa, parasites
Chemical Harmful/corrosive/toxic chemicals,
biologicals (plant toxins, venoms,
microbial toxins)
Other Cancerous cells/tumours, immune
response (autoimmunity,
hypersensitivity), ischaemia (lack
of blood supply and subsequent
death of the tissue)
- Acute = fast
- Chronic = long term
- Chronic: lymphocytes are the adaptive arm of the immune response
Main processes
- Vascular changes
o Leads to swelling as well as the influx of the mediators that are there to
clean up afterwards
- Chemotactic factors released
o Factors that attract cells into the site those are the cellular infiltrate and
effectors
Cellular infiltration and effector functions
- Fever
o Is triggered by certain cytokines, can help to control the proliferation of
certain bacteria that do not survive in hot environments
- Healing
- Many of these initiate and/or participate in the inflammatory process; not
separate events, all come together
Vascular changes
- When you have tissue damage, the first phase of inflammation is the fluidic
phase get fluid going into the site due to vascular changes like vasodilation
and increased vascular permeability
o Can also have direct effect if there is an injury to a blood vessel, this will
directly allow passage of fluids and other mediators into the site
- Image: bottom picture of blood vessel in an inflamed tissue get increase in
blood flow with dilation of the vessels, leakage of plasma proteins into the space
oedema, also get migration of neutrophils out of the vessels and into the
space which happens quite rapidly
Cellular response
- Margination: leukocytes move to the edge of the blood vessels rather than
central flow
- Rolling and adhesion: mediated by receptors on the leukocytes and
corresponding receptors on activated endothelial cells selectins and integrins
- Extravasation and migration: leukocytes pass between endothelial cells and
move to the site of inflammatory stimuli
- Image (above): if you have a stimulus (green), it could in its own right trigger
this or may have response to the bacteria that are part of that insult, releases
vasoactive and chemotactic factors which lead to extravasation of the cells
which migrate to the site. That occurs through a process that involves
margination, rolling and adhesion and extravasation and migration
- When the cells start to marginate they move to the edge of the blood vessels
they are associating more closely with the endothelial cells (cells that line the
blood vessels)
- Then they start rolling along these endothelial cells, that is mediated by
receptors on those cells called selectins and integrins
- These endothelial cells are not passive, are active participants in this process,
are expressing receptors that allow the neutrophils and macrophages to bind on
and go across the vascular wall
- Then the leukocytes can move between endothelial cells in order to move closer
to the site of inflammation stimuli (squeeze in between the endothelial cells)
- Image: timing of this overall process is initially get oedema because of the
vascular changes that occur very quickly (swelling, entry of fluids into the space
fluidic phase), then get entry of neutrophils first responders, then get
monocytes and macrophages coming in later
- Emigration from blood into tissue process of rolling, adherence and emigration
between endothelial cells (inflammation)
- Margination and rolling
o Margination where they start moving near the edge of the vascular wall
o They start rolling along until they get a firm adhesion because there is
formation of complementary binding between the receptors on the cells
and the receptors on the endothelial wall
This is a very important for how the lymphocytes and leukocytes get
around they need to know where to go this process of receptor
expression by cells and endothelial cells is how leukocytes know
where they need to go, if they dont see a complementary receptor
then they will stay in the blood stream, if they do, then they know
they need to go to that tissue site
o This process of squeezing between is called diapediesis
o Then get chemotactic movement along the gradient of the chemokines
This is where phagocytosis comes into play
Endothelial cells
Tissue repair
1 2 3
Summary
- Complement is a series of plasma proteins that act in a cascade that can lead to
formation of the MAC and other inflammatory/immune mediators
- Three main pathways that can activate the complement cascade: lectin,
classical and alternative
- Interaction between innate and adaptive immunity; complement the ability of
antibody to lyse bacteria
- Five cardinal signs of inflammation
- Vascular and cellular changes in inflammation
Humoral Immunity
Review
- Image:
o Innate: have barriers, have enzymes and antimicrobial substances being
produced, have complement that is acting in the innate immune process,
have different cell types
o Unless an animal has a minor issue, will get induction of adaptive immune
responses; innate immune responses are really important though
because, if there were congenital deficiencies in many of the innate
pathways any organism with those is highly susceptible to recurrent
bacterial and other types of infections (very important on a daily basis)
o However, adaptive immune responses are where evolution of the immune
response has moved on well developed
Learning outcomes
Bacterial genome
Epitopes
- From the development of those mature B and T cells, get activation of particular
cell types that are the responder cells of the adaptive immune response
o For B cells, these are the plasma cells: produce lots of antibodies
releasing it into the blood stream or to the site
o Can also form memory: memory cell populations come after there is
activation of a particular immune response long-lived cell population
that is produced in order for this to occur
o There are also different antibody isotypes: not just one type of antibody
different as they have different function (e.g. might need to be in blood,
tissues, lumen of the gut structures need to be different)
- Like a Y
shape
because the
majority of
antibody
isotypes are
quite flexible
- These arms
(Fab
fragment)
can move to
adjust for a
different type
of antigen
- The arms have variable regions that are the parts of the antibody that can
recognise an antigen
- Constant regions: are referred to as the FC region, they do not vary this is
important because it is the FC portion of the antibody that can be recognised by
other cells of our immune system (e.g. phagocytes have receptors for that FC
portion)
- Have two chains: heavy and light chains called this because they have
different molecular weights
o In the antibody pictured above, it is symmetrical, so the heavy chains are
the same in any particular antibody and the light chains are the same
So an antibody, either arm, will recognise exactly the same epitope
of an antigen
- Top image:
o When you have the secreted
form of an antibody, it loses
a chunk which is this
transmembrane region and
cytoplasmic tail
o The membrane bound
versions are slightly larger,
they are inserted into the
membrane of the B cells and
include the transmembrane
region and cytoplasmic tail
o The secreted versions are
truncated
o Can make exactly the same
molecule but for different
purposes
- Bottom image:
o Can see there are some
that look very much like
the basic structure of
an antibody and others
look like joined
together versions of
that basic structure
o All mammalian species
have these represented
classes of antibody, but
there are species
specific variations
Some species
only have certain
subclasses,
others have a
slightly different
structure
IgM
IgG
IgA
IgD
- Expressed only on the surface of mature,
nave (havent seen anything before, not
activated) B cells
- Signal 1: the direct binding of the B cell receptor (an antibody that is bound to
the surface of the B cell) and the signal (whatever the antigen is)
- B cells need two more signals to become fully activated; need to bind to T cells
that are specific for that same antigen (via MHC II and costimulatory molecules
e.g. CD4), and
o CD40 and CD40L (ligand): CD40 is on the surface of the B cell and CD40L
is on the surface of the T cell. Once there is binding of both and the MHCII
to the T cell receptor, can then get activation of the B cell
o T cell is also providing cytokines, mediators that are helping the B cell to
respond and these are IL-4 etc.
Clonal expansion
- What are the effector mechanisms of the antibody? What do they actually do?
o Can bind directly to viruses and bacteria, and neutralise them
o Neutralisation of toxins: binding to a toxic production produced by a
bacteria for example, can then remove it and stop it from having that
toxic effect
o Agglutination: so can get removal by phagocytes
o Precipitation of soluble antigens: so can get removal by phagocytes
Phagocytes: have an FC receptor, they can see/recognise that tail
portion of the antibody and realise this needs to be internalised
(form of opsonisation)
o Interaction with the complement system: getting opsonisation or
triggering directly of cell lysis by the MAC
- There are multiple ways antibodies can have an effect
Monoclonal antibodies
- Before monoclonal
antibodies, could
generate a type of
immune response
that was non-specific
(very difficult to see
differences between
cell populations to
see these
differences, need to
be able to see what
their surface
receptor population
looks like
- To see this, need a
monoclonal antibody,
something that recognises a particular receptor on a cell type
- Produced by the fusion of splenocytes from a mouse that has been immunised
against whatever you want to look for
- Wil have a B cell response, these B cells will be present in the spleen of the
mouse some time later with a kind of cancer cell (myeloma cell line)
- If these are fused together, can create a very long lived cell lined called a
hybridoma
- Through a process of clonal selection of the individual cells and growing those
up, can get a clone that produces just the type of antibody that you want to
produce
- These clones can be grow in in the lab, can be frozen and then brought back up
- Allows production of many of a particular antibody to anything you want it to
recognise
- Some are even being used as biological therapeutics
Serological testing
Summary