The immune sytem

Phagocytes: Origin & Mode of Action

 Phagocytes are white blood cells that are produced continuously in the bone marrow
 They are stored in the bone marrow before being distributed around the body in the
blood
 They are responsible for removing dead cells and invasive microorganisms
 They carry out what is known as a non-specific immune response
 There are two main types of phagocyte, each with a specific mode of action. The two
types are:
o Neutrophils
o Macrophages
 As both are phagocytes, both carry out phagocytosis (the process of recognising and
engulfing a pathogen) but the process is slightly different for each type of phagocyte

Neutrophils

 Neutrophils travel throughout the body and often leave the blood by squeezing through
capillary walls to ‘patrol’ the body tissues
 During an infection, they are released in large numbers from their stores
 However, they are short-lived cells
 Mode of action:
o Chemicals released by pathogens, as well as chemicals released by the body cells
under attack (eg. histamine), attract neutrophils to the site where the pathogens
are located (this response to chemical stimuli is known as chemotaxis)
o Neutrophils move towards pathogens (which may be covered in antibodies)
o The antibodies are another trigger to stimulate neutrophils to attack the pathogens
(neutrophils have receptor proteins on their surfaces that recognise antibody
molecules and attach to them)
o Once attached to a pathogen, the cell surface membrane of a neutrophil extends
out and around the pathogen, engulfing it and trapping the pathogen within
a phagocytic vacuole
o This part of the process is known as endocytosis
o The neutrophil then secretes digestive enzymes into the vacuole (the enzymes are
released from lysosomes which fuse with the phagocytic vacuole)
o These digestive enzymes destroy the pathogen
o After killing and digesting the pathogens, the neutrophils die
o Pus is a sign of dead neutrophils
 The stages of phagocytosis, as carried out by a neutrophil

Macrophages

 Macrophages are larger than neutrophils and are long-lived cells

 Rather than remaining in the blood, they move into organs including the lungs, liver,
spleen, kidney and lymph nodes
 After being produced in the bone marrow, macrophages travel in the blood as monocytes,
which then develop into macrophages once they leave the blood to settle in the various
organs listed above
 Mode of action:
o Macrophages play a very important role in initiating an immune response
o Although they still carry out phagocytosis in a similar way to neutrophils, they do
not destroy pathogens completely
 o They cut the pathogens up so that they can display the antigens of the pathogens
on their surface (through a structure called the major histocompatibility complex)
o These displayed antigens (the cell is now called an antigen-presenting cell) can
then be recognised by lymphocytes (another type of white blood cell)

Antigens
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Antigens, Self & Non-Self

 Every cell in the human body has markers that identify it

 Microorganisms (both pathogenic and non-pathogenic), such as bacteria and viruses, also
have their own unique markers
 These markers are called antigens (which are macromolecules) and they allow cell-to-cell
recognition
 Antigens are found on cell surface membranes, bacterial cell walls, or the surfaces of viruses
o Some glycolipids and glycoproteins on the outer surface of cell surface membranes
act as antigens
 Antigens can be either self antigens or non-self antigens:
o Antigens produced by the organism’s own body cells (those that the immune
system does not recognise as foreign antigens) are known as self antigens
o Self antigens do not stimulate an immune response
o Antigens not produced by the organism’s own body cells (those that the immune
system recognises as being foreign eg. the antigens found on pathogenic bacteria
and viruses or if a person receives a different blood type during a transfusion) are
known as non-self antigens
o Non-self antigens stimulate an immune response

Primary Immune Response

Primary Immune Response (Advanced)

 Lymphocytes are another type of white blood cell

 They play an important part in the specific immune response
 They are smaller than phagocytes
 They have a large nucleus that fills most of the cell
 They are produced in the bone marrow before birth
 There are two types of lymphocytes (with different modes of action). The two types of
lymphocytes are:
o B-lymphocytes (B cells)
o T-lymphocytes (T cells)

B-lymphocytes

 B-lymphocytes (B cells) remain in the bone marrow until they are mature and then spread

through the body, concentrating in lymph nodes and the spleen
 Millions of types of B-lymphocyte cells are produced within us because as they mature the
genes coding for antibodies are changed to code for different antibodies
 Once mature, each type of B-lymphocyte cell can make one type of antibody molecule
 At this stage, the antibody molecules do not leave the B-lymphocyte cell but remain in the
cell surface membrane
 Part of each antibody molecule forms a glycoprotein receptor that can combine specifically
with one type of antigen

The maturation of B-lymphocytes – by the time a child is born, it will have millions of
different types of B-lymphocytes, each with a specific antibody receptor

 When an antigen enters the body for the first time, the small numbers of B-lymphocytes
with receptors complementary to that antigen are stimulated to divide by mitosis
 This is known as clonal selection
 As these clones divide repeatedly by mitosis (the clonal expansion stage) the result is large
numbers of identical B-lymphocytes being produced over a few weeks
 During an immune response, these B-lymphocytes then form two types of cell:
o Some of these B-lymphocytes become plasma cells that secrete lots of antibody
molecules (specific to the antigen) into the blood, lymph or linings of the lungs and
the gut
o These plasma cells are short-lived (their numbers drop off after several weeks) but
the antibodies they have secreted stay in the blood for a longer time
o The other B-lymphocytes become memory cells that remain circulating in the blood
for a long time
o This response to a newly encountered antigen is relatively slow and is known as
a primary immune response
 During a primary immune response, B-lymphocytes form two types of cell

T-lymphocytes

 Immature T-lymphocytes leave the bone marrow to mature in the thymus

 Mature T-lymphocytes have specific cell surface receptors called T cell receptors
 These receptors have a similar structure to antibodies and are each specific to one antigen
The maturation of T-lymphocytes – some become helper T cells and others become killer T
cells

 T-lymphocytes are activated when they encounter (and bind to) their specific antigen that is
being presented by one of the host’s cells (host cells being the human’s own cells)
 This antigen-presenting host cell might be a macrophage or a body cell that has been
invaded by a pathogen and is displaying the antigen on its cell surface membrane
 These activated T-lymphocytes (those that have receptors specific to the
antigen) divide by mitosis to increase in number (similar to the clonal selection and clonal
expansion of B-lymphocytes)
 These T-lymphocytes differentiate into two main types of T cell:
o helper T cells
o killer T cells
 Helper T cells release cytokines (hormone-like signals) that stimulate B-lymphocytes to
divide and develop into antibody-secreting plasma cells. Some helper T cells secrete
cytokines that stimulate macrophages to increase their rates of phagocytosis
 Killer T cells attach to the antigens on the cell surface membranes of infected cells
and secrete toxic substances that kill the body cells, along with the pathogen inside

Helper T cells and killer T cells carry out different functions during an immune response

Memory Cells & Immunity

Memory Cells & Long-Term Immunity

 During an immune response, B-lymphocytes form two types of cell: plasma

cells and memory cells
  Memory cells form the basis of immunological memory – the cells can last for many
years and often a lifetime
 There are two types of immune response:
o Primary immune response (responding to a newly encountered antigen)
o Secondary immune response (responding to a previously encountered antigen)

Primary immune response

 When an antigen enters the body for the first time, the small numbers of B-lymphocytes
with receptors complementary to that antigen are stimulated to divide by mitosis
 This is known as clonal selection
 As these clones divide repeatedly by mitosis (the clonal expansion stage) the result is large
numbers of identical B-lymphocytes being produced over a few weeks
 Some of these B-lymphocytes become plasma cells that secrete lots of antibody molecules
(specific to the antigen) into the blood, lymph or linings of the lungs and the gut
 These plasma cells are short-lived (their numbers drop off after several weeks) but the
antibodies they have secreted stay in the blood for a longer time
 The other B-lymphocytes become memory cells that remain circulating in the blood for a
long time
 This response to a newly encountered pathogen is relatively slow

Secondary immune response

 If the same antigen is found in the body a second time, the memory cells recognise the
antigen, divide very quickly and differentiate into plasma cells (to produce antibodies) and
more memory cells
 This response is very quick, meaning that the infection can be destroyed and
removed before the pathogen population increases too much and symptoms of the disease
develop
 This response to a previously encountered pathogen is, relative to the primary immune
response, extremely fast
During a secondary immune response, memory cells that remained in the blood divide very
quickly into plasma cells (to produce antibodies) and more memory cells

 T-lymphocytes also play a part in the secondary immune response

 They differentiate into memory cells, producing two main types:
o Memory helper T cells
o Memory killer T cells
 Just like the memory cells formed from B-lymphocytes, these memory T cells remain in the
body for a long time
 If the same antigen is found in the body a second time, these memory T cells become active
very quickly

Antibodies and vaccination

Antibodies
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Antibodies: Structure & Functions

Structure

 Antibodies are globular glycoproteins called immunoglobulins

 Antibodies have a quaternary structure (which is represented as Y-shaped), with two
‘heavy’ (long) polypeptide chains bonded by disulfide bonds to two ‘light’ (short)
polypeptide chains
 Each polypeptide chain has a constant region and variable region
 The constant regions do not vary within a class (isotype) of antibodies but do vary between
the classes. The constant region determines the mechanism used to destroy the antigens
o There are 5 classes of mammalian antibodies each with different roles
 The amino acid sequence in the variable regions of the antibodies (the tips of the “Y”) are
different for each antibody. The variable region is where the antibody attaches to the
antigen to form an antigen-antibody complex
 At the end of the variable region is a site called the antigen-binding site. Each antigen-
binding site is generally composed of 110 to 130 amino acids and includes both the ends of
the light and heavy chains
 The antigen-binding sites vary greatly giving the antibody its specificity for binding
to antigens. The sites are specific to the epitope (the part of the antigen that binds to the
antibody)
 A pathogen or virus may therefore present multiple antigens different antibodies need to be
produced
 The ‘hinge’ region (where the disulfide bonds join the heavy chains) gives flexibility to the
antibody molecule which allows the antigen-binding site to be placed at different angles
when binding to antigens
o This region is not present in all classes of antibodies
 A model of the generalised structure of an antibody molecule

Function

 Antibodies are produced by B-lymphocytes

 Antibodies bind to specific antigens that trigger the specific immune response. Every antigen
has one antibody
 Antigens include pathogens and their toxins, pollen, blood cell surface molecules and the
surface proteins found on transplanted tissues
 Antibodies are divided into five major classes (isotypes), each with a different role
 The function of antibodies differ:
o Antibodies can combine with viruses and toxins of pathogens (e.g. bacteria) to block
them from entering or damaging cells
o Antibodies can act as anti-toxins by binding to toxins produced by pathogens (e.g.
the bacteria that cause diphtheria and tetanus) which neutralises them making them
harmless
o Antibodies can attach to bacteria making them readily identifiable to phagocytes,
this is called opsonisation. Once identified, the phagocyte has receptor proteins for
the heavy polypeptide chains of the antibodies, which enables phagocytosis to occur
o Antibodies can attach to the flagella of bacteria making them less active, which
makes it easier for phagocytes to do phagocytosis
o Antibodies act as agglutinins causing pathogens carrying antigen-antibody
complexes to clump together (agglutination). This reduces the chance that the
pathogens will spread through the body and makes it possible for phagocytes to
engulf a number of pathogens at one time
o Antibodies (together with other molecules) can create holes in the cell walls of
pathogens causing them to burst (lysis) when water is absorbed by osmosis
 The functions of antibodies vary according to which type of antigen they act on

Making Monoclonal Antibodies

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The Hybridoma Method

 Monoclonal antibodies are artificially produced antibodies produced from a single B cell
clone
 The hybridoma method is a method used to make monoclonal antibodies (Mabs)
 The method enables large quantities of identical antibodies to be produced
 The hybridoma method solved the problem of having B cells that could divide by mitosis but
not produce antibodies and plasma cells that could produce antibodies but not divide
 This method was established in the 1970s
 Monoclonal antibodies bind antigens, in the same way naturally produced antibodies do
 They are produced by injecting mice with an antigen that stimulates the production of
antibody-producing plasma cells
 Isolated plasma cells from the mice are fused with immortal tumour cells, which result
in hybridoma cells
 These hybrid cells are grown in a selective growth medium and screened for the production
of the desired antibody
 They are then cultured to produce large numbers of monoclonal antibodies
 Monoclonal antibodies have multiple applications to include diagnostics, treating disease,
food safety testing and pregnancy testing
 The hybridoma method is used to produce monoclonal antibodies

Uses of Monoclonal Antibodies

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Use of Monoclonal Antibodies
Diagnostic uses of monoclonal antibodies

 Monoclonal antibodies can be used diagnostically for:

o Pregnancy tests
o Diagnosing HIV
o Detecting the presence of pathogens such as Streptococcus bacteria
o Distinguishing between Herpes I and Herpes II
o Blood typing before transfusions and tissue typing before transplants
o Detecting the presence of antibiotics in milk
o Detecting cancer cells
 Monoclonal antibodies can also be used to locate the position of blood clots for patients
thought to have deep vein thrombosis. This occurs by:
o Injecting a mouse with human fibrin (the main protein found in blood clots)
o This activates the plasma cells to produce antibodies against fibrin
o These cells are collected from the mouse spleen
o The plasma cells are then fused with tumour cells forming hybridomas that produce
antifibrin antibodies
o To detect where the antibodies are binding to fibrin molecules, a radioactive
chemical (producing gamma radiation) is attached to the antibodies making them
radioactively labelled
o A gamma-ray camera is used to detect where these radioactively labelled antibodies
have attached to a fibrin molecule, hence indicating where blood clots can be found
 Generally monoclonal antibodies are used only once
 Another example of the diagnostic use of monoclonal antibodies – test for HIV

Therapeutic uses of monoclonal antibodies

 Therapeutically monoclonal antibodies have multiple applications to include:

o Treatment for the rabies virus, (which can be potentially fatal), by injecting purified
antibodies
o The prevention of transplanted organ rejection, achieved by intervening with the T
cells involved in the rejection process
o Autoimmune therapies for allergic asthma and rheumatoid arthritis; here
monoclonal antibodies are able to bind and deactivate factors involved in the
inflammatory response
o Treatment for diseases caused by the overproduction or inappropriate production
of B-cells (eg. leukaemia, multiple sclerosis and myasthenia gravis); the antibody
(rituximab) binds to cell surface receptor proteins on B-cells (not plasma cells) and
causes the death of the cells
o Prevention of blood clotting following angioplasty procedures; here monoclonal
antibodies bind to receptors on the platelet surface thereby inhibiting fibrinogen
from binding and subsequent clotting from ensuing
o Targeted treatment of breast cancer; Herceptin (trastuzumab) is a monoclonal
antibody used to treat breast cancer, it recognises receptor proteins on the surface
of cancer cells and binds to them allowing the immune system to identify and
destroy them
o Treatment of melanoma (a type of skin cancer); the antibody (ipilimumab) binds to a
protein produced by T-cells (whose role is to reduce the immune response) which
results in the immune system remain active against the cancer cells

 Using monoclonal antibodies as a treatment requires multiple administrations and this can
cause problems
 Initially the monoclonal antibodies were produced by mice, rabbits or other laboratory
animals (as these were easier to produce), however this triggered an immune response
when they were introduced to humans
 Scientists have largely overcome this by:
o Genetically modifying the antibody polypeptide chains so that the amino acid
sequences are now human not mouse or rabbit sequences
o Altering the type and position of the sugar groups (antibodies are glycoproteins)
attached to the heavy polypeptide chains to reflect those found on human
antibodies

Types of Immunity
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Types of Immunity
Active immunity

 Active immunity is acquired when an antigen enters the body triggering a specific immune
response (antibodies are produced)
 Active immunity is naturally acquired through exposure to microbes or artificially
acquired through vaccinations
 The body produces memory cells, along with plasma cells, in both types of active immunity
giving the person long-term immunity
 In active immunity, during the primary response to a pathogen (natural) or to a vaccination
(passive), the antibody concentration in the blood takes one to two weeks to increase. If the
body is invaded by the same pathogen again or by the pathogen that the person was
vaccinated against then, during the secondary response, the antibody concentration in the
blood takes a much shorter period of time to increase and is higher than after the
vaccination or first infection
 The primary and secondary response to the same antigen

Passive immunity

 Passive immunity is acquired without an immune response. Antibodies are not produced by
the infected person
 As the person’s immune system has not been activated then there are no memory cells that
can produce antibodies in a secondary response. If a person is reinfected they would need
another infusion of antibodies
 Depending on the disease a person is infected with (eg. tetanus) they may not have time to
actively acquire the immunity, that is, there is no time for active immunity. So passive
immunity occurs either artificially or naturally
 Artificial passive immunity occurs when people are given an injection / transfusion of the
antibodies. In the case of tetanus this is an antitoxin. The antibodies were collected from
people whose immune system had been triggered by a vaccination to produce tetanus
antibodies
 Natural passive immunity occurs when:
o Foetuses receive antibodies across the placenta from their mothers
o Babies receive the initial breast milk from mothers (the colostrum) which delivers a
certain isotype of antibody (IgA)

Comparing Active & Passive Immunity Table

How Vaccines work
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How Vaccines Work

 A vaccine is a suspension of antigens that are intentionally put into the body to

induce artificial active immunity. A specific immune response where antibodies are
released by plasma cells
 There are two main types of vaccines:
o Live attenuated
o Inactivated
 Vaccines are administered either by injection or orally (by mouth). When a person is given a
vaccine they have been given a vaccination
 The vaccinations given by injection can be into a vein or muscle
 Vaccinations produce long-term immunity as they cause memory cells to be created. The
immune system remembers the antigen when reencountered and produces antibodies to it,
in what is a faster, stronger secondary response

 Vaccines can be:

o Highly effective with one vaccination giving a lifetime’s protection (although less
effective ones will require booster / subsequent injections)
o Generally harmless as they do not cause the disease they protect against because
the pathogen is killed by the primary immune response
 Unfortunately there can be problems with vaccines:
o People can have a poor response (eg. they are malnourished and cannot produce
the antibodies – proteins or their immune system may be defective)
o A live pathogen may be transmitted (e.g. through faeces) to others in the population
(ideally enough number of people are vaccinated at the same time to give herd
immunity)
o Antigenic variation – the variation (due to major changes) in the antigens of
pathogens causes the vaccines to not trigger an immune response or diseases
caused by eukaryotes (eg. malaria) have too many antigens on their cell surface
membranes making it difficult to produce vaccines that would prompt the immune
system quickly enough
o Antigenic concealment – this occurs when the pathogen ‘hides’ from the immune
system by living inside cells or when the pathogen coats their bodies in host proteins
or by parasitising immune cells such as macrophages and T cells (eg. HIV) or by
remaining in parts of the body that are difficult for vaccines to reach (eg. Vibrio
cholerae – cholera, remains in the small intestine)
 The principles underpinning vaccinations were discovered by Edward Jenner in the 1700s
when he developed the first smallpox vaccine

Live attenuated vaccines

 Live attenuated vaccines contain whole pathogens (e.g. bacteria and viruses) that have been
‘weakened’
 These weakened pathogens multiply slowly allowing for the body to recognise the antigens
and trigger the primary immune response (plasma cells to produce antibodies)
 These vaccines tend to produce a stronger and longer-lasting immune response
 They can be unsuitable for people with weak immune systems as the pathogen may divide
before sufficient antibodies can be produced
  An example of this type of vaccine is the MMR (Measles, Mumps and Rubella)

Inactivated vaccines

 Inactivated vaccines contain whole pathogens that have been killed (‘whole killed’) or small
parts (‘subunit’) of the pathogens (eg. proteins or sugars or harmless forms of the toxins –
toxoids)
 As inactivated vaccines do not contain living pathogens they cannot cause disease, even for
those with weak immune systems
 However these vaccines do not trigger a strong or long-lasting immune response like the live
attenuated vaccines. Repeated doses and / or booster doses are often required
 Some people may have allergic reactions or local reactions (eg. sore arm) to inactivated
vaccines as adjuvants (eg. aluminium salts) may be conjugated (joined) to the subunit of the
pathogen to strengthen and lengthen the immune response
 An example of a whole killed vaccine is polio vaccine
 An example of a toxoid subunit vaccine (where inactivated versions of the toxins produced
by pathogens are used) is Diphtheria