Chapter 11

Immunity
 Defense against disease

There are two types of defense mechanism

1. External defense system

2. Internal defense system

 External defense system
 The first line of defense includes physical and chemical barriers that are

always ready and prepared to defend the body from infection.

 External defense systems include our skin, tears, mucus, saliva, stomach

acid, cilia (small hairs), and helpful bacteria in our bowel.

 Internal defense system
 Internal defense systems activate when the external defenses fail and

something bad finds its way into our bodies.

 It involves two types of white blood cells.

1. phagocytes and

2. lymphocytes.
 Immune system: the body’s internal defense system.

 Antigen (pathogen): a substance that is foreign to the body and stimulates an

immune response. An antigen can be proteins, glycoproteins, glycolipids and

polysaccharides, and the toxins and waste materials

 Self: refers to substances produced by the body that the immune system does not

recognize as foreign, so they do not stimulate an immune response.

 Non-self: refers to any substance or cell that is recognized by the immune system

as being foreign and will stimulate an immune response.

 Antibody: a glycoprotein (immunoglobulin) made by specialized

lymphocytes in response to the presence of a specific antigen; each type of

antibody molecule has a shape that is complementary to its specific antigen.

 Immune response: the complex series of responses of the body to the

entry of a foreign antigen; it involves the activity of lymphocytes and

phagocytes.

 Immunity: is the protection against disease provided by the body’s

internal defense or immune system.

 Phagocytes

 Phagocytes are cells that protect the body by ingesting harmful foreign particles, bacteria,

and dead or dying cells.

 Phagocytes are produced throughout life in the bone marrow. They are stored there before

being distributed around the body in the blood.

 They are scavengers, removing any dead cells as well as invasive microorganisms.

 They carry out non-specific immune responses.

 There are two types of phagocytes:

1. Neutrophils or

2. Monocytes/macrophages
 Neutrophils
 About 60% of the white cells in the blood are neutrophils.

 They travel throughout the body, often leaving the blood by squeezing through the

walls of capillaries to move through the tissues engulfing any pathogens that they find.

 During an infection, neutrophils are released in large numbers from their stores, but

they are short-lived cells (5.4 days).

 Monocytes/macrophages
 They are made in the bone marrow and circulate in the blood as monocytes.

 When there is any attack from bacteria or viruses, they eventually leave the blood.

 When they leave the blood they become macrophages.

 Monocytes and macrophages can live for several months.

 They play a crucial role in initiating immune responses, since they do not destroy

pathogens completely, but cut them up to display antigens that can be

recognized by lymphocytes.
 Phagocytosis
 A process by which certain living cells called phagocytes ingest or engulf other cells

or particles.

 If pathogens invade the body and cause an infection, some of the cells under attack

respond by releasing chemicals such as histamine.

 These, together with any chemicals released by the pathogens themselves, attract

passing neutrophils to the site. This movement towards a chemical stimulus is called

chemotaxis.
 The neutrophils destroy the pathogens by phagocytosis in two ways.

1. Directly engulf the pathogen or

2. Engulf the pathogen with the help of antibodies.

Directly engulf the pathogen

 When the neutrophil directly attaches to the pathogens, the neutrophil’s cell

surface membrane engulfs the pathogens and traps them within a phagocytic

vacuole in a process called endocytosis.

 After that, lysosome fuse with the phagocytic vacuole(phagosome) which is

known as phagolysosome and release enzymes that breakdown the pathogens.

 Neutrophils have a short life: they die after killing and digesting some pathogens.

 Dead neutrophils often collect at a site of infection to form pus.

Engulf the pathogen with the help of antibodies.
 Pathogens are captured by antibodies.
 Neutrophils have receptor proteins on their surfaces that recognize antibody
molecules and attach to them.
 When the neutrophil attaches to the pathogens, the neutrophil’s cell surface
membrane engulfs the pathogens and traps them within a phagocytic vacuole in a
process called endocytosis.
 Lysosomes fuse with the phagocytic vacuoles releasing enzymes that break down
the pathogens.
 Macrophages
 Macrophages are larger than neutrophils and are long-lived cells.
 Macrophages play a role in initiating a specific immune response as follows:
1. Macrophages carry out phagocytosis similar to neutrophils, but they do not
destroy pathogens completely during the digestion stage of the process.
2. They cut the pathogens up so that they can display the antigens of the
pathogens on their surface.
3. Macrophages that present antigens in this way are known as antigen presenting
cells.
4. These displayed antigens can then be recognised by lymphocytes, another group
of white blood cells.
 Lymphocytes
 Lymphocytes are smaller than phagocytes.
 They have a large nucleus that fills most of the cell.
 There are two types of lymphocytes, which are produced in the bone marrow.
1. B-lymphocytes and
2. T-lymphocytes
 Only mature lymphocytes can carry out immune responses.

 Lymphocytes, unlike phagocytes, act against specific pathogens.

 Each lymphocyte contains a set of genes that codes for the production of a

particular type of receptor.

 Immune responses depend on B and T cells interacting with each other to give

an effective defense.
 B-lymphocytes (B cells)
 B cells remain in the bone marrow until they are mature and then spread
throughout the body, concentrating in lymph nodes and the spleen. Although
some continue to circulate in the blood.

 Lymph nodes are small, bean-shaped

organs that support the immune system by
trapping foreign agents and killing them.
 Spleen filters blood and presents foreign
particles (antigens) to the lymphocytes.
 Many different types of B cells develop in each of us, perhaps as many as 10 million.
 While B cells are maturing, the genes that code for antibodies are changed in a
variety of ways to code for different antibodies.
 Each cell then divides to give a small number of cells that are able to make the
same type of antibody. Each small group of identical cells is called a clone.
 Once mature, each different B-lymphocyte cell has a different antibody molecule on
its surface.
 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 receptor that can combine with
complementary antigens from pathogens; these receptors are known as antibody
receptors or B cell receptors (BCR), as they are made of antibodies.
Clonal selection: is a process by which the body

produces B and T cells to respond to infections.

These cells each have unique receptors that

allow them to identify specific pathogens.

Clonal expansion/ proliferation: the increase

in number of specific clones of lymphocytes by

mitosis during an immune response.

B cells can be divided into two types.

1. Plasma cells give immediate protection by the secretion of specific antibody

molecules very quickly – up to several thousand a second. These plasma cells do

not live long: after several weeks their numbers decrease.

2. Memory B cells give rapid and enhanced responses to secondary exposure to

the same pathogen. These cells last for many years, often a lifetime.
 B lymphocytes in the specific immune response
 When non-self antigens enter the body, the B lymphocytes with antibody receptors that are
complementary to the new antigen, come into contact with the new antigen.
 The B lymphocytes bind to their complementary antigens and are stimulated to divide by mitosis.
 This process of finding and stimulating the correct lymphocytes is known as clonal selection.
 The B lymphocytes divide repeatedly by mitosis, producing many cloned B lymphocytes with
complementary antibody receptors.
 The cloning of B lymphocytes at this stage is known as clonal expansion.
 The cloned B lymphocytes develop into:
1. Plasma cells secrete antibodies that are complementary to the antigens that initiate the immune
response.
2. Memory cells remain in the blood, providing immunity during secondary infection; these are
known as B memory cells.
Action of B-lymphocytes
Different ways of action by
antibodies against antigen
 T-lymphocytes
 T-lymphocytes (T cells) leave the bone marrow and mature in the thymus.

 The thymus is a gland that lies in the chest just beneath the sternum.

 Mature T cells have specific cell surface receptors called T-cell receptors (TCR)

similar to that of antibodies, and they are each specific to one antigen.
 T lymphocytes in the specific immune response
 T cells are activated when they recognize the antigen on another cell of the host.
 Sometimes this cell is a macrophage that has engulfed a pathogen and cut it up to expose the
pathogen’s surface molecules, or it may be a body cell that has been invaded by a pathogen for
recognition by T-lymphocytes.
 The display of antigens on the surface of cells in this way is known as antigen presentation.
 The T cells that have receptors complementary to the antigen respond by dividing by mitosis to
increase the number of cells. T cells go through the same stages of clonal selection and clonal
expansion as clones of B cells.
There are two main types of T cells:

1. T-helper cells and

2. T-killer cells (also known as T-cytotoxic cells).

Action of T-helper cells
 When T-helper cells bind to antigens, they release cytokines (cell-signaling
molecules) that stimulate appropriate B cells to divide, develop into plasma
cells, and secrete antibodies. Also, they produce memory T cells.
 Some T-helper cells secrete cytokines that stimulate macrophages to carry out
phagocytosis more vigorously.
 Some other T-helper cells secrete cytokines that stimulate T-killer cells to
divide by mitosis and to differentiate by producing vacuoles full of toxins.
Action of T-killer cells
 T-killer cells search the body for cells that have become invaded by pathogens.
 When T-killer cells recognize the antigens, attach themselves to the surface of
infected cells, and secrete toxic substances such as hydrogen peroxide, killing
the body cells and the pathogens inside. Also, they produce memory T cells.
 Memory T-helper cells and memory T-killer cells are produced, which remain in
the body and become active very quickly during the secondary response to
antigens.
 Primary immune response: the first immune

response to a specific antigen. This response is

slow because, at this stage, there are very few B

cells that are specific to the antigen.

 Secondary immune response: the second and

any subsequent immune responses to a specific

antigen. This response is faster because there

are now many memory cells, which quickly

divide and differentiate into plasma cells.

 Why primary response is slower?

 Since it is the first time the immune system has encountered the antigen, the numbers

of T and B cells with the correct membrane receptors present in the blood will be low.

 It will take time for the correct lymphocytes to be activated and to divide and for

different cell types to develop.

 It can take several days before plasma cells develop and are able to start producing

antibodies against an antigen.

 This is the reason why an infected person will experience symptoms of the disease the

first time they contract it.

 Why secondary response is much larger and faster than the primary response?
 The primary response produces memory cells which remain in the blood after an infection is
over.
 The presence of memory cells means that a person is said to be immune to the pathogen.
Memory cells can remain in the blood for many years, so the immunity gained from a primary
response can be long-term.
 When the immune system encounters the same antigen again in the future, it will launch a
secondary immune response which will be much faster and stronger than the primary response.
 Memory cells are present in larger quantities than the mature lymphocytes at the start of the
primary response, so the correct memory cells can detect an antigen, activate, and divide by
mitosis much more quickly.
 Antibodies are produced more quickly and in larger concentrations in a secondary response.
 This will often eliminate the pathogen before the infected person can show symptoms.
 Antibody structure

 Antibodies are globular glycoproteins called immunoglobulins.

 Antibodies have a Y-shaped quaternary structure with two ‘heavy’ polypeptide chains

bonded by disulfide bonds to two ‘light’ polypeptide chains

 Antibodies have a constant region and a variable region.

 The constant regions do not vary within a class of antibodies but do vary between the

classes. The constant region determines the mechanism used to destroy antigens.

 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 the antigen-binding site. Each antigen-binding site

is generally composed of 110 to 130 amino acids.

 Antigen-binding sites vary greatly, giving the antibody its specificity for binding to

antigens.

 Antibodies bind to a region of the antigen called the epitope.

 Antibodies also have a hinge region which gives flexibility to the antibody molecule.

 This allows the antigen-binding site to be placed at different angles when binding to

antigens.
 Types of Immunity

There are two types of immunity.

1. Active immunity

2. Passive 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 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.
 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, there are no memory cells
that can produce antibodies in a secondary response.
 Artificial passive immunity occurs when people are given an
injection/transfusion of the antibodies. The antibodies were collected from
people whose immune systems had been triggered by a vaccination.
 Natural passive immunity occurs when: Fetuses receive antibodies across the
placenta from their mothers and babies receive the initial breast milk from mothers
which delivers a certain type of antibody.
 Vaccines
 A vaccine is a preparation containing antigens which is used to stimulate an immune
response artificially.
 It may contain:
1. a whole live microorganism (weak form),
2. a dead one,
3. a harmless version (known as an attenuated organism),
4. a harmless form of a toxin (known as a toxoid), or
5. a part of pathogen.

 Vaccines are either given by injection into a vein or muscle, or taken orally (by mouth).
How Vaccines Work?
 Vaccination helps the immune system to mimic the infection procedure.
 Live vaccines are very effective because microorganisms reproduce often
rather slowly, so the immune system can mimic their infection procedure.
 Less effective are those vaccines that do not mimic an infection because they
are made from dead bacteria or from viruses that do not reproduce inside cells
of the body.

 Vaccines that are highly effective, one dose may give a lifetime’s protection.
 Less effective vaccines need booster injections to stimulate secondary
responses that give enhanced protection.
 MMR vaccine for measles, mumps, and rubella at about nine months of age

and a second dose at any time after a minimum interval of four weeks. The

second dose is usually given between 15 months and 4 years of age.

 BCG vaccine to protect against TB (Tuberculosis).

 Problems with vaccines

 People can have a poor response. For example: peoples who are malnourished

and cannot produce the antibodies (which are proteins) or their immune system

may be defective.

 A live pathogen may be transmitted (e.g. through faeces) to others in the

population.
 Vaccination to Control Disease

1. Herd immunity

2. Ring immunity
 Herd immunity
 It occurs when a significant portion of a population becomes immune to

an infectious disease and the risk of spread from person to person

decreases.

 If most of the populations are immune to a specific disease, it is unlikely that

the pathogen will be transmitted to those who do not have immunity.

 Because herd immunity interrupts the transmission cycle in a population.

So that those who are susceptible are very unlikely to be infected by the

pathogens concerned.
 Ring immunity
 Vaccinating all those people in contact with a person infected with a
specific disease to prevent transmission in the immediate area.
 Monoclonal antibodies

 Monoclonal antibodies (also called moAbs or mAbs) are proteins made in

laboratories by hybridoma cells that act like proteins called antibodies in our

bodies.

 The cell produced by this fusion of a plasma cell and a cancer cell (tumor cell/

myeloma cell) is called a hybridoma (cancer of the plasma cells).

 Hybridoma cells will have the property of plasma cell antibody production and

cancer cells ability to continuously division.

Monoclonal antibodies production procedure
1. A mouse or other organism is injected with an antigen that will stimulate the
production of the desired antibody.
2. Plasma cells that secrete antibodies are then taken from the spleen of the
mouse.
3. These plasma cells are fused with cancer cells to produce hybridoma cells.
4. The hybridoma cells then continuously divide and produce different types of
antibodies.
5. Each antibody is then tested for the desired antibody.
6. A clone of hybridoma cells is produced which only makes the desired antibody.
 Use of Monoclonal Antibodies

Monoclonal antibodies can be used diagnostically for:

 Pregnancy tests

 Diagnosing HIV

 Detecting the presence of pathogens such as Streptococcus bacteria

 Distinguishing between Herpes I and Herpes II virus

 Blood typing before transfusions and tissue typing before transplants

 Detecting the presence of antibiotics in milk

 Detecting cancer cells

 For example, Mabs can be used to locate the position of blood clots in the body of a
person thought to have a deep vein thrombosis (DVT).
Process of Mab for blood clotting:

 Inject a mouse with human fibrin to produce antibodies.

 The mouse makes many plasma cells that secrete the antibody against fibrin.

 The plasma cells are fused with cancer cells to form hybridomas that secrete the

anti-fibrin antibody.

 A radioactive chemical that produces gamma radiation is attached to each

antibody molecule to make a radioactively labelled Mab.

How can we apply Mab?

 The labelled antibodies are introduced into the patient’s blood.

 As the Mabs are carried around the body in the bloodstream, they bind to any fibrin

molecules with which they come into contact.

 The radioactivity emitted by these labelled antibodies is used to detect where they

are in the body.

 A gamma-ray camera is used to detect the exact position of the antibodies in the

person’s body.

 The position of the labelled Mabs indicates the position of any blood clots.
Another example of monoclonal antibodies in the diagnosis

 Monoclonal antibodies can be used in an ELISA (enzyme-linked immunosorbent assay)

test to diagnose infectious disease, by detecting the presence of particular antigens in

the blood.

1. The monoclonal antibody is immobilized (attached) on the surface of a small glass well.

The liquid to be tested (for example, blood plasma) is added to the well. If the antigens

are present, they will bind to the antibodies.

2. Unbound antigens are then washed away. The antigens stay tightly bound to the

antibodies and those aren’t bound will be washed away.

Continue…

3. Now another solution containing the same monoclonal antibodies which

bind with the antigens already attached to the antibodies is added to the well.

These antibodies also have a reporter enzyme combined with them. The well is

again washed out, so the enzymes will all be washed away unless they have bound

with the antigens.

4. The substrate of the enzyme is then added. If the enzyme is present — which

is only the case if the antigen being investigated was present — then the substrate

is changed to a coloured substance by the enzyme.

ELISA plate
Another example of the diagnostic use of monoclonal antibodies - test for HIV
Some Mabs have had significant success in treatments.

These monoclonals are:

1. trastuzumab,

2. ipilimumab,

3. infliximab and

4. rituximab.
 Trastuzumab (HerceptinTM)

 Trastuzumab is used in the treatment of some breast cancers.

 Trastuzumab binds to a receptor protein that is produced in abnormal

quantities in the cell surface membranes of some breast cancers and

marks them out for destruction by the immune system.

 Ipilimumab
 It is a therapy for melanoma, a type of skin cancer.

 It also works by activating the immune system but in a different way than

trastuzumab.

 Ipilimumab binds to a protein produced by T cells, the role of which is to

suppress (reduce) the immune response.

 By blocking the action of the protein, an immune response can be maintained

against the cancer cells.

 Infliximab
 It is used to treat rheumatoid arthritis.

 This condition involves a protein secreted by T-lymphocytes that causes

damage to the cartilage in joints.

 Infliximab binds to the protein and blocks its action.

 Rituximab
 It is used to treat diseases in which there is an overproduction or inappropriate

production of B-lymphocytes.

 Rituximab is used to control B-lymphocytes.

 This Mab binds to a cell surface receptor protein found on the surface of B cells.

 Following binding to B cells, rituximab causes a variety of changes that lead to the

death of the B cells.

 New B cells are made in the bone marrow, and these replacement cells may not be

cancerous.
 Problems and solutions for Mabs
 The antibodies are produced in laboratory animals.
 When introduced into humans, they trigger an immune response because they are
foreign (non-self) and act as antigens.
This problem has now been largely overcome in two ways:
1. altering the genes that code for the heavy and light polypeptide chains of the
antibodies so that they code for human sequences of amino acids rather than mouse
or rabbit sequences.
2. changing the type and position of the sugar groups that are attached to the heavy
chains to the arrangement found in human antibodies.