Immunity

11.1.1 Phagocytes
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)
Exam Tip
The vacuole formed around a bacterium once it has been engulfed by a phagocyte is called a
phagosome. A lysosome fuses with the membrane of the phagosome (to form a
phagolysosome) and releases lysozymes (digestive enzymes) to digest the pathogen.

11.1.2 Antigens
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

11.1.3 Primary Immune Responses

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
Lymphocytes in the Primary Immune Response Diagram
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
11.1.4 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
Exam Tip
Immunological memory (made possible by memory cells) is the reason why
catching certain diseases twice is so unlikely. For example, there is only one
strain of the virus that causes measles, and each time someone is re-infected
with this virus, there is a very fast secondary immune response so they do not
get ill.However, some infections such as the common cold and influenza are
caused by viruses that are constantly developing into new strains. As each
strain has different antigens, the primary immune response (during which we
often become ill) must be carried out each time before immunity can be
achieved.

11.2.1 Antibodies
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
Exam Tip
You must know that each antibody will have a different variable region with an
antigen-binding site that matches one antigen or toxin produced by a
pathogen. The antigen-binding site (and therefore the antibody)
is specific to one antigen.

11.2.2 Making Monoclonal Antibodies

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)
 o Monoclonal antibodies bind antigens, in the same way naturally
produced antibodies do
 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
 The hybridoma method involves
o Injecting mice with an antigen that stimulates the production of
antibody-producing plasma cells
o Isolated plasma cells from the mice are fused with
immortal tumour cells, which result in hybridoma cells
 The fusion of plasma and tumour cells can be assisted with
the use of fusogens such as polyethylene glycol or an
electric current
o These hybrid cells are grown in a selective growth medium and
screened for the production of the desired antibody
o 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
Exam Tip
Remember monoclonal antibodies are produced from a hybridoma cell - a cell
formed by the fusion of plasma cells and tumour (cancer) cells, which divide
continuously therefore producing large quantities of a wanted antibody.

11.2.3 Uses of Monoclonal Antibodies

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
Exam Tip
Know specific examples of how monoclonal antibodies can be used as a
diagnostic tool and for treatment. You can use a well-annotated diagram to
explain how monoclonal antibodies can be used as a diagnostic tool.

11.2.4 Types of Immunity

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 (artificial), 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

Exam Tip
Active immunity is when the body produces the antibodies whereas in passive
immunity the body is given the antibodies.
11.2.5 How Vaccines work

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
Exam Tip
Remember vaccines trigger the primary immune response (T helper cells
trigger B plasma cells to secrete specific antibodies) which leads to the
production of memory cells which will give a faster and greater (higher
concentration of antibodies) during the secondary response.

11.2.6 Vaccination to Control Disease

Vaccination to Control Disease

 With the exception of the great success story surrounding the
eradication of Smallpox following a ten year global initiative in 1980 no
other pathogen has been eradicated globally since
 Smallpox was able to be eradicated because a ‘live attenuated’ vaccine
was used against the only strain of the virus. There was also a
programme of surveillance, contact tracing and ‘ring’ vaccinations
 There are many safe and effective vaccines that do exist against many
pathogens and these have managed to push a number of childhood
diseases to the verge of extinction
  Vaccines against such diseases as mumps, chicken pox and whooping
cough are administered to children as part of an immunisation schedule
and they successfully confer immunity
 As a result many childhood diseases are kept at low levels within
populations due to herd immunity
 Herd immunity arises when a sufficiently large proportion of the
population has been vaccinated (and are therefore immune) which
makes it difficult for a pathogen to spread within that population, as
those not immunised are protected and unlikely to contract it as the
levels of the disease are so low
 Although most vaccinations are given to children there are some
vaccines that are provided at later stages in life, for example vaccinations
for tuberculosis (TB) and Hepatitis B are offered to frontline medical
workers who have a higher risk of coming into contact with such diseases
in the hospital setting;
Travellers may be advised to take particular vaccines if travelling to areas where
certain diseases are endemic such as Yellow Fever in parts of Africa