Chapter 11

Immunology
By the end of this chapter you should be able to:
a describe the mode of action of phagocytes; b describe the roles of mast cells and histamine production, complement, and phagocytes as antigen-presenting cells; c define the term immune response; d compare the origin and maturation of B- and T-lymphocytes, including the types of T-cells and their functions, and B-cells and their functions; e distinguish between the humoral and the cellmediated immune responses; f explain the role of T- and B-memory cells in long-term immunity; g relate the molecular structure of a typical antibody molecule to its function, including specificity; h distinguish between active and passive immunity, natural and artificial immunity; i j explain the role of vaccination in providing immunity; state what is meant by a monoclonal antibody;

k describe the use of monoclonal antibodies in diagnosis and treatment, including pregnancy testing, and the anticancer drug MabThera.

Parasites and pathogens

Infectious diseases are ones that we can catch from someone else, such as a cold, TB, malaria and HIV/AIDS. These diseases are caused by pathogens. A pathogen can be defined as a microorganism that causes disease. Pathogens are a kind of parasite. A parasite is an organism that lives in a very close relationship with another organism, called its host, and does it harm. The parasite gains from the relationship. So all pathogens are parasites, but not all parasites are pathogens. For example, you might have lice living in your hair, but they are not causing a disease so they are not pathogens. A well-adapted parasite or pathogen does not kill its host. The parasite or pathogen is most likely to survive, and produce offspring that can move to a new host, if its host survives long enough for this to happen. Most of the infectious diseases that have been around for a long time, such as colds, measles and TB, either do not kill us or do not kill us quickly.

Pathogens belong to one of four different groups of microorganisms viruses, bacteria, fungi and protozoa. (Some may argue that viruses are not organisms at all.) Table 11.1 lists some examples of diseases caused by each of these groups.

The immune response

We have numerous defences against invasion of our bodies by pathogens. The first line of defence is to stop them getting in at all. If they do gain access, then the immune system comes into action. The way in which white blood cells respond when pathogens enter the body is called the immune response. Several types of white blood cells (leucocytes) are able to recognise foreign cells or molecules that enter the body. In other words, they can distinguish self from non-self. The immune response is the way in which the immune system responds to the presence of non-self cells or molecules in the body. 221

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Pathogen human immunodeficiency virus (HIV) adenovirus Mycobacterium Tinea pedis Plasmodium

Type of microorganism virus virus bacterium fungus protozoan

Disease caused acquired immune deficiency syndrome (AIDS) colds tuberculosis (TB) athletes foot malaria

Table 11.1 Causes of some infectious diseases.

Primary lines of defence

The best line of defence against pathogens is to prevent them from getting established in the body. Skin is impermeable to most pathogens, although there are a few viruses, such as the ones that cause warts, that can penetrate unbroken skin. We have our own flora of harmless bacteria that live on healthy skin, but most pathogenic bacteria cannot survive there, partly because lactic acid and fatty acids secreted from sweat glands and sebaceous glands provide a pH that is too low for them. However, the common bacterium Staphylococcus aureus can thrive even on undamaged skin, and it often infects hair follicles and sebaceous glands. The normal bacterial flora living on our body surfaces can help to prevent infection by other microorganisms. For example, the bacteria that normally live in the vagina keep the pH low by secreting lactic acid. If a person takes antibiotics, these bacteria may be killed. Then the pH of the vagina rises, and this may allow other microorganisms, such as the fungus that causes thrush, Candida, to multiply to a much greater population density than usual. If skin is damaged for example, by cuts or extensive burns then the way is open for bacteria to get into the underlying tissues. Blood clotting helps to seal wounds rapidly, until a more permanent repair is produced by mitosis of the cells surrounding the wound. A blood clot forms when soluble, globular fibrinogen is converted to the insoluble, fibrous protein fibrin. This forms a mesh of strands across the wound in which platelets stick and red blood cells get trapped, thus preventing further loss of blood or entry of pathogens. Moist body surfaces, such as the surface of the eyes and mouth, are bathed in fluids which have some bactericidal action. An enzyme called lysozyme is present in saliva and tears, and this enzyme can damage and destroy many bacteria. Semen contains a bactericide called spermine; milk contains a bactericidal enzyme called lactoperoxidase. The hydrochloric acid secreted into the stomach is very effective in destroying bacteria and other pathogens ingested in food. Mucus helps to protect the digestive and respiratory tracts from infection. It acts as a barrier so that bacteria cannot make contact with the epithelial cells lining the walls of the tubes. Mucus is produced by goblet cells, which are part of the epithelium. A layer of cells containing goblet cells is sometimes known as a mucous membrane (but dont confuse this membrane with a plasma membrane of a cell). In the trachea and bronchi, the mucus is swept upwards to the back of the throat by cilia and then swallowed. Coughing and sneezing help to expel mucus containing microorganisms from the trachea and bronchi. If the mucus is swallowed, the acid and enzymes in the stomach destroy any bacteria trapped in it.

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Phagocytes
If pathogens do get through the bodys outer defences, they may be destroyed by patrolling phagocytic white blood cells. The types of white blood cells known as neutrophils and macrophages are phagocytes. They engulf and digest foreign particles of almost any type or size (Figure 11.1). They crawl around within almost every part of the body for example, over the surfaces of the alveoli in the lungs. Neutrophils are found in the blood, where they make up about 60% of the white blood cells. They do not live very long, often dying after they have taken in and destroyed bacteria, and so new neutrophils are constantly being made in the bone marrow. They move around actively, and frequently leave the blood and patrol parts of the body where invaders may be found. Macrophages also leave the blood. (Indeed, when they are actually in the blood they are given a different name monocytes.) They are present
1 Phagocytic white blood cell moves towards a pathogen.

in especially large numbers in the liver, where they are known as Kupffer cells. They also line the passages through which lymph flows inside lymph nodes and are found on the inside of the alveolar walls. Unlike neutrophils, they are quite long-lived, tending to survive after taking in foreign particles. They break the particles up into their component molecules and place some of these molecules in their plasma membranes. Cells that do this are called antigen-presenting cells. By doing this, they display the molecules to other cells of the immune system, helping these cells to identify the invaders and be able to destroy them. This role is described more fully on pages 226228. For phagocytosis to take place, the microorganisms must first adhere to the plasma membrane of the phagocyte. This process is helped by a group of proteins called complement (page 228) which are always present in the blood plasma, and also by chemicals called cytokines, which are produced by other white blood cells in response

2 Phagocytosis takes place.

3 Lysosomes join with the vacuole (phagosome) and the pathogen is killed and digested. lysosome phagosome

4 Any chemicals that are not absorbed into the cell are egested.

Figure 11.1

Phagocytosis. 223

Chapter 11: Immunology to the presence of particular antigens. Cytokines make phagocytes more efficient at killing any microorganisms that they have engulfed. The way in which phagocytes deal with invading cells or other foreign material is a non-specific response. Each phagocyte can attack and destroy any type of non-self material.

One type of antibody is held in the plasma membrane acting as a receptor for a specific antigen. The same antibody can be secreted from the cell in quantity.

Lymphocytes
Lymphocytes are relatively small white blood cells. They are of two types, B-lymphocytes and T-lymphocytes. These two types look identical, and differ only in their functions. B-lymphocytes are so-called because they develop in the bone marrow, while T-lymphocytes need to spend time in the thymus gland during a persons childhood to become properly developed. This gland is found in the neck. It disappears by the time a person becomes a teenager. Lymphocytes are stimulated into action when they come into contact with molecules called antigens. Invading bacteria and viruses are recognised as foreign because they carry or produce antigens that are different from any of our own molecules. Antigens may be free or they may be part of a bigger structure, such as the cell wall of a bacterium. We have a huge number of different kinds of lymphocytes in our blood. Each one is capable of recognising and responding to one particular antigen. The response of lymphocytes to non-self molecules is therefore known as a specific response. As they mature, lymphocytes produce small quantities of particular glycoproteins called antibodies (page 233). We have perhaps a million different kinds of lymphocytes, each kind producing an antibody which is slightly different from other antibodies. At this stage, the antibodies are placed into the plasma membranes of the lymphocytes (Figure 11.2). Here, the antibodies act as receptors, able to bind with a particular antigen if this should appear in the body. If bacteria enter the body, there is a good chance that some of the lymphocytes will have receptors that bind with antigens on the surface of the bacteria. If so, then a response is triggered. B-lymphocytes and T-lymphocytes respond differently. 224

Figure 11.2 A lymphocyte can produce one specific type of antibody. How B-lymphocytes respond to antigens Most B-lymphocytes will spend all their lives without anything happening to them at all, because they never meet their particular antigen. But if a B-lymphocyte does encounter an antigen which binds to the receptors in its plasma membrane, it is triggered into action. It could simply meet this antigen in the blood, or it could meet it as it is being displayed in the plasma membrane of an antigen-presenting cell (APC) such as a macrophage (Figure 11.3). You can imagine the macrophages sitting in the lymph channels inside a lymph node, holding out the antigens they have discovered so that the lymphocytes will see them as they pass by. The B-lymphocyte responds by dividing repeatedly by mitosis. A large number of genetically identical cells is formed a clone of the stimulated lymphocyte. The process of the B-lymphocyte binding with its specific antigen is sometimes called clonal selection, and its division to form a clone of genetically identical cells is called clonal proliferation or clonal expansion. Some of these cells differentiate into plasma cells. These cells develop extra protein-making machinery more endoplasmic reticulum, more ribosomes and more Golgi apparatus. They rapidly synthesise more and more molecules of their particular antibody and release them by exocytosis. It has been estimated that a plasma cell can produce and release more than 2000 antibody

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bacterium phagocytosis by an antigen-presenting cell (APC), e.g. macrophage antigen processed and displayed

The B-lymphocyte displays an antibody specific to the antigen on the bacterium.

The B-lymphocyte meets its specific antigen either on a macrophage or on the bacterium.

The B-lymphocyte divides to produce many plasma cells, which all secrete antibodies.

Figure 11.3 B-lymphocyte response to antigen. molecules per second. Perhaps as a direct result of this tremendous rate of activity, plasma cells do not live long, mostly disappearing after only a few weeks. The antibodies are secreted into the blood and so are carried to all parts of the body. They bind with the antigens on the invading bacteria, which results in the destruction of the bacteria as we shall see on pages 232233. Other cells in the clone produced by the original B-lymphocytes division do not secrete antibodies. Instead, they remain as memory cells. These cells live for a long time, and remain circulating in the blood long after the invading bacteria have all been destroyed. They are capable of responding very quickly if the same type of bacterium enters the body again. How T-lymphocytes respond to antigens T-lymphocytes, like B-lymphocytes, are activated if and when their particular antigen binds with the specific glycoproteins that are held in their plasma membranes. T-lymphocytes, however, normally only respond to their antigen if they find it in the plasma membrane of another cell. This could be a macrophage that is displaying some of the molecules from a pathogen that it has taken up. Or it could be molecules on a body cell that has been invaded by a virus, and has placed virus particles in its plasma membrane as a help signal (Figure 11.4). There are several types of T-lymphocytes, including T-helper cells and T-killer cells. A particular T-helper cell with the complementary receptor binds to the antigen that it has found. It then divides to form a clone of itself. The cloned T-helper cells then begin to secrete chemicals called cytokines. These chemicals stimulate other cells to fight against the invaders. For example, they may stimulate macrophages to carry out phagocytosis, or they may stimulate B-lymphocytes specific to this antigen to divide rapidly and become plasma cells. They also help to stimulate appropriate T-killer cells. T-killer cells actually destroy the cell to which they have become bound. A body cell displaying virus particles will be destroyed by T-killer cells. This is the only way of destroying the viruses it cant be done without destroying the cell in which they are multiplying. The T-killer cells destroy 225

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A macrophage (APC) ingests, processes and displays an antigen.

The T-lymphocyte meets the specific antibody on a macrophage or another APC.

T-helper cells The T-helper or T-killer lymphocyte displays on its plasma membrane an antibody specific to the antigen. The T-lymphocyte divides. T-killer cells T-helper cells secrete cytokines to stimulate phagocytic cells and B-lymphocytes. A cell infected by virus displays the antigen on its surface.

T-killer cells kill the cells they attach to.

Figure 11.4 T-lymphocyte response to antigen. the infected cell by secreting chemicals such as hydrogen peroxide. The T-killer cells are our main defence against viral diseases. We have seen that T-lymphocytes, like B-lymphocytes, divide to form clones when they meet their own particular antigen (Figure 11.5). While most of these cells act as helper cells or killer cells, some of them remain in the blood as memory cells. These, like the memory cells formed from B-lymphocytes, help the body to respond more quickly and effectively if this same antigen ever invades again. SAQ 1 With reference to the way in which they respond to antigens, suggest why T-lymphocytes are more effective than B-lymphocytes in dealing with infection by a virus. SAQ 2 Match each of these words with its definition below. antibody, antigen, pathogen, parasite, B-lymphocyte, neutrophil, macrophage a a type of white blood cell that divides to produce plasma cells, which secrete antibodies b a phagocytic white blood cell with a multilobed nucleus and granular cytoplasm c a molecule that is recognised by lymphocytes as being foreign to the body d an organism that lives in close association with a host, and does it harm. e a microorganism that causes disease f a glycoprotein secreted by some white blood cells, which binds to specific antigens g a phagocytic white blood cell that is relatively large, and which tends to be found in tissues such as the lungs, rather than in the blood

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B-cells Specific binding B-lymphocyte with antibody in its plasma membrane binds to complementary antigen. antigen

T-cells Antigen presentation An APC (macrophage) ingests, processes and presents antigen.

Clonal selection and proliferation Stimulated B-lymphocyte divides many times.

Specific binding T-helper lymphocyte or T-killer lymphocyte binds to complementary antigen on an APC.

Memory cells These survive for a long time.

Plasma cells These secrete large amounts of antibody.

Clonal selection and proliferation One clone is stimulated and this T-lymphocyte divides many times.

either

If antigen appears later, the memory cells are stimulated, divide and produce many plasma cells very quickly.

T-killer lymphocytes These bind to cells presenting the complementary antigen.

T-helper lymphocytes These secrete cytokines which stimulate phagocytic cells and other lymphocytes. memory cells

memory cells T-killer cells bind to cells presenting the complementary antigen and kill them.

cell killed

Figure 11.5 Summary of B-lymphocyte and T-lymphocyte actions. 227

Chapter 11: Immunology SAQ 3 To answer this question, you will need to think back to your work on cells. An experiment was carried out to follow what happens inside plasma cells as they make and secrete antibodies. Some cells were cultured in a solution containing amino acids which had been labelled with a radioactive marker. The radioactivity in the Golgi body, endoplasmic reticulum and ribosomes was then measured over the next 40 minutes. The results are shown in the graph.
40 Radioactivity / arbitrary units 30 20 10 0 ribosomes cisternae of endoplasmic reticulum Golgi body

10

20 30 Time / minutes

40

a In which order did the amino acids move through the three organelles? Use the results shown in the graph to justify your answer. b Using your own knowledge, describe what happened to the amino acids in each organelle. c Suggest why the peak values for the radioactivity in the ribosomes and the endoplasmic reticulum are the same, whereas the peak value for the Golgi body is lower. (There may be more than one possibility.) d Suggest how the amino acids would have been taken up into the cell at the beginning of the experiment. e Describe how the antibody molecules would be secreted from the cell.

Complement
Complement is a collection of small proteins (more than 25 different ones) that are always present in the blood plasma. It was first discovered in 1895, and was given this name because it helps, 228

or complements, the activity of antibodies and phagocytes. Complement is very important in fighting bacterial infections. Many of the proteins that make up complement are precursors of enzymes. When a piece of their molecule is removed, they become active. Once one of them has been activated in this way, it acts as a catalyst for the activation of another complement protein. This becomes a cascade process, in which one small action (whatever activates the first protein) ends up having a very large effect on a large number of protein molecules (Figure 11.6). There is more than one way in which the cascade can be initiated. Firstly, when an antibody binds to an antigen, one of the complement proteins can bind to the antibody. This changes the shape of the complement protein, activating it and setting off the cascade. Alternatively, a different complement protein can bind directly with a pathogen (or any other non-self surface). Once again, this changes its shape and starts off the cascade. The result of either of these events is the production of various proteins that can help to destroy invading microorganisms. There are three ways in which they do this. Opsonisation Some of the proteins produced when the complement cascade is activated bind to the surface of bacteria, coating them with a layer of protein called opsonin. Phagocytic cells have receptors which bind to opsonin, and this stimulates them to engulf and destroy the coated bacterium. Attracting macrophages and other cells to the site of infection Some of the newly produced complement proteins drift away from the place where they were formed, into the tissue fluid and blood. Their presence attracts phagocytes and other white blood cells, which move towards the site. (Cell movement in the direction of a chemical stimulus is an example of chemotaxis.) This is important in the inflammatory response. Destroying foreign cells A third kind of complement protein directly destroys the cells that stimulated its production, by making holes in their plasma membranes.

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Complement is a group of short-lived soluble proteins always present in blood plasma. Complement is activated by contact with antibody bound to antigen, or with a foreign surface. Activation of complement triggers a cascade which results in the production of active proteins. activation inactive enzyme active enzyme active enzyme active complement protein

The active proteins help destroy foreign cells.

Opsonins coat a foreign cell which encourages phagocytosis.

inactive enzyme a complement protein

inactive complement protein

Phagocytes are attracted to the area by active complement protein.

Foreign cells are destroyed by active complement protein.

Figure 11.6 Complement.

The inflammatory response

If a pathogen gets into a particular area of your body and begins to multiply, it is no use having your phagocytes and lymphocytes spread all over the body you need them to be concentrated in the danger area. The process that brings this about is called the inflammatory response, and it results in inflammation (Figure 11.7). Imagine, for example, that a thorn has penetrated deep under your skin. Bacteria on the thorn begin to multiply. The presence of antigens on the bacteria, and your own damaged tissues, activate the complement system. Chemicals are released that increase the blood supply to the area and make the capillaries more permeable. This brings more phagocytes and lymphocytes to the infected tissues. Phagocytes are attracted to the area by the chemicals, and they crawl out of the blood into the infected tissues. The extra blood supply makes the area look red, and the leakage of fluid from the blood makes it swollen. If all goes well, your body will win the battle against the pathogens, and the swelling and redness will subside as the infection is brought under control. Sometimes, a thick white mixture of dead bacteria, lymphocytes and phagocytes builds up, known as pus.

Mast cells
Mast cells are cells that are found in all tissues, generally lying close to the walls of blood vessels and nerves. Their cytoplasm is packed full of granules (Figure 11.8), which contain numerous chemicals, especially histamine and heparin. We know a lot about mast cells because they are very much involved in allergies and auto-immune diseases. Both of these result from the immune system behaving inappropriately, causing illness. However, it is also thought that mast cells do have a useful role to play, probably in helping the immune system to fight intestinal worms and other parasites. Mast cells must be activated before they begin to do anything. There are three main ways in which this happens. They may respond directly to injury. This could be physical, or it could be caused by toxic chemicals such as alcohol. The membranes of mast cells have receptors that bind tightly to a type of antibody called IgE, so each mast cell is completely coated with IgE molecules. If the protein that fits into the IgE molecule binds with them, the IgE molecules become linked together, and this activates the mast cell. Unfortunately, this

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In an inflammation, blood supply to an injured or infected area is increased. Capillaries in the area become more permeable allowing more chemicals to leave the blood plasma and enter the area. White blood cells crawl out of the capillaries by chemotaxis. tissue damaged by a cut

inflammation

Tissue damage and the presence of antigens from bacteria, for example, act as a stimulus. capillary

There is greater leakage of plasma containing soluble substances e.g. complement, clotting factors, antibodies.

Activities taking place at the site of injury may include: blood clotting, antibody-antigen interaction, phagocytosis and killing of pathogens. White blood cells are attracted to the area and leave through holes in the capillary wall.

Arterioles supplying capillaries dilate, increasing blood flow into capillaries. This causes the capillaries to dilate.

Clotting and immune responses are activated.

Figure 11.7 Inflammation. often happens not in response to a potentially dangerous pathogen, but to a harmless antigen such as a protein in the surface of a pollen grain, or on a cell in a peanut. These substances that should be harmless, but that act as antigens and bring about a strong and inappropriate immune response, are known as allergens. Activated complement proteins can also activate mast cells. An activated mast cell releases the contents of its granules. These include histamine and several cytokines. These cause an acute inflammatory reaction, in which blood vessels dilate, smooth muscle in airways contracts, rashes appear on the skin and tissues swell as fluid accumulates in them. In a severe allergic reaction, mast cells all over the body release their contents at the same time, causing a massive inflammatory response that can be life-threatening. Several diseases are the result of a misdirected attack of the immune system on a persons own tissues, and these are known as auto-immune diseases. Mast cells are known to play a major part

nucleus

vesicles (granules) that contain histamine

mitochondria

Figure 11.8 A coloured electronmicrograph of a mast cell (12000). 230

Chapter 11: Immunology in many of these, including rheumatoid arthritis (in which the joints become inflamed) and multiple sclerosis (in which the myelin sheaths of neurones are destroyed). Despite much research, there is still no clear picture of what causes these diseases to develop. There does seem to be some genetic component, because auto-immune diseases may have a tendency to run in families. However, there is also an environmental component, because the development of an auto-immune disease often seems to follow infection by a virus.

epitope variable region hinge region light chain disulphide links polypeptides polysaccharide chains heavy chain constant region

Humoral and cell-mediated responses

Early studies of the immune system suggested that the body had two different ways of attacking pathogens. One involved cells the phagocytes and T-lymphocytes and was called the cell-mediated response. The other involved chemicals, especially antibodies produced by B-lymphocytes, and was called the humoral response. It is now known that, in reality, there are constant and complex interactions between cells and chemicals, as you will have appreciated from what you have read earlier in this chapter.

Figure 11.9 The structure of an antibody molecule. There are several different kinds of them, given names such as IgG and IgA. Each antibody contains a variable region that can bind specifically with a particular antigen. We have millions of different antibodies with different variable regions. The particular part of the antigen that is recognised by the immune system, and to which the antibody attaches, is called an epitope. When an antibody molecule meets its specific antigen, it binds with it. The effect that this has depends on what the antigen is, and on what type of immunoglobulin has bound to it. Some antibodies directly neutralise the antigen for example, by binding with a toxin produced by a bacterium. Others may encourage phagocytes to destroy the pathogen, sometimes by making the pathogens clump together. Yet others may stop pathogens getting a foothold on body surfaces, by preventing them from attaching to cells or tissues (Figure 11.10 and Figure 11.11).

HIV/AIDS and the immune system

The human immunodeficiency virus infects a particular group of T-helper cells called CD4+ cells, and also some types of macrophages. In the disease AIDS, the presence of the virus causes a reduction in the numbers of CD4+ cells. This can be because the virus itself destroys the cell as it reproduces inside it and bursts out from it; or because other T-lymphocytes recognise that the CD4+ cell is infected, and attack and destroy it. The reduction in numbers of the CD4+ cells greatly weakens the ability of the immune system to respond to infection, and it is this that causes the symptoms of AIDS.

Antibodies
Antibodies are glycoproteins. Their molecules contain chains of amino acids, and also sugar units. Figure 11.9 shows the structure of an antibody molecule. Antibodies are also known as immunoglobulins.

How immunity develops

When a pathogen first enters the body, there will be only a few lymphocytes with receptors that fit into its antigens. It takes time for these lymphocytes to encounter and bind with these pathogens. It takes more time for them to divide to form clones, and for the B-lymphocytes to secrete enough 231

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antibody

toxin produced by bacteria pathogenic bacteria Antigen binding stimulates phagocytic white cells to ingest and kill the bacterium. Toxins released by pathogens can be neutralised by antibodies.

An antibody can neutralise a bacterium if the antibody binds to a chemical necessary for pathogenicity.

Figure 11.10

How antibodies neutralise bacteria and bacterial toxins. If the same pathogen invades again, these memory cells can mount a much faster and more effective response. More antibodies can be produced more quickly, usually destroying the pathogen before it has caused any illness. The response to the first invasion of the pathogen is called the primary response (Figure 11.12). Subsequent invasions generate a secondary response. You can see that the secondary response happens more quickly, and produces many more antibodies. This is why we usually become immune to a disease if we have had it once.

antibody with several sites that can bind to antigen pathogenic bacterium with antigens on its surface

The antibody makes the bacteria clump together (agglutinate). Agglutinated bacteria don't move around as much and are more readily ingested by phagocytic white cells.

Active and passive immunity

The kind of immunity described on the previous pages is a type of active immunity. The immune system has been stimulated to make a particular type of antibody, and can produce this same one more quickly and in larger quantity if it is exposed to the same pathogen again. The immunity has developed naturally, so it is a type of natural immunity. Another way in which active immunity can develop is by vaccination. This involves injecting the antigen into the body (page 224). It may, for example, be in the form of viruses that have been made harmless, or as an inactivated toxin from a bacterium. The body responds in the same way as it would if invaded by the living pathogen, producing memory cells which will make the person immune to the disease if they should ever encounter it. This way of acquiring active

Figure 11.11 bacteria.

How antibodies agglutinate

antibodies to destroy the pathogens, or for enough T-lymphocytes to be produced to be able to destroy all the cells that are infected by them. During this delay, the pathogens have the opportunity to divide repeatedly, forming large populations in the body tissues. The damage that they cause, and toxins that they may release, can make the person ill. It may be several days, or even weeks, before the lymphocytes get on top of the pathogen population and destroy it. However, if the body survives this initial attack by the pathogen, memory cells will remain in the blood long after the pathogen has been destroyed. 232

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Primary response Concentration of antibody in the blood Concentration of antibody in the blood 1 2 Time / weeks weeks or years later

Secondary response

infection

infection

1 2 Time / weeks

Figure 11.12 Primary and secondary responses to antigen. immunity is not natural, so it is a form of artificial immunity (Figure 11.13). A young babys immune system takes time to develop. In the uterus, the foetus obtains antibodies from the mothers blood, across the placenta. After birth, the baby will continue to receive them in the mothers milk, if she decides to breastfeed. These ready-made antibodies help the baby to fight off pathogens. The baby has immunity to the same diseases as the mother. Because the babys body has received ready-made antibodies, rather than making them itself, this is said to be passive immunity. It has happened
Active immunity Immunity developed after contacting pathogens inside the body.

naturally, so it is an example of natural immunity too. Passive immunity can also be provided by injections. This is not a natural way of gaining immunity, so it is another example of artificial immunity. For example, if someone goes to the emergency department of a hospital with a cut that may have dirt in it, they may need to be protected against the bacterium that causes tetanus, Clostridium tetani. It is too late for a vaccination, because by the time the persons immune system responds, the bacterium could have multiplied and caused the fatal illness tetanus. Instead, the
Passive immunity Immunity provided by antibodies or antitoxins provided from outside the body.

Natural infection

Artificial injection of live or attenuated pathogen

Natural antibodies from a mother in breast milk or across the placenta

Artificial injection of antibodies or antitoxin

Figure 11.13 Active and passive immunity. 233

Chapter 11: Immunology person will be given an injection of antitoxin. The antitoxin will bind to the toxin produced by the bacteria, rendering it harmless. Antitoxins can also be given in this way to destroy toxins introduced into the body through bites from poisonous animals, such as spiders or snakes. Passive immunity does not last as long as active immunity. No lymphocytes have been stimulated to make clones of themselves, so no memory cells have been formed. Passive immunity lasts only as long as the antibodies or antitoxins last. The body actually sees them as being foreign, and they will be removed and destroyed quite quickly by cells in the liver and spleen. attenuated (weakened) form of the bacterium or virus that causes the disease, while others contain a modified toxin produced by them. When the vaccine enters the body, lymphocytes that recognise the antigen respond to it as if they had encountered live bacteria or viruses. They form clones of plasma cells, which secrete antibodies, and also memory cells. In most cases, a second booster dose of the vaccine is given later on. This raises the antibody level much higher than the first dose, and helps to ensure that protection against the antigen lasts for some time (Figure 11.14).

Monoclonal antibodies
We have seen that there is a huge number of different antibodies that can be made by human B-lymphocytes, and that each lymphocyte can make only one kind. In the 1970s, researchers wanted to be able to obtain large amounts of one particular antibody at a time, so that they could study it without interference from all the other antibodies that are usually present in a mammals blood. Their aim was to produce a large clone of a particular type of B plasma cell, all secreting identical antibodies, known as monoclonal antibodies. There is one problem in achieving this B-lymphocytes that divide to form clones of plasma cells do not secrete antibodies, and plasma

Vaccination
Vaccination is an excellent way of preventing a person from acquiring an infectious disease. The larger the proportion of people who are vaccinated in a population, the lower the chance that anyone even those who have not been vaccinated will get that disease. This is called herd immunity. For most diseases, at least 8085% of the population need to be vaccinated to achieve herd immunity. Vaccination involves giving a person a dose of a preparation that will cause the immune system to react as though an antigen from a pathogenic organism has entered the body. Most vaccinations are given by injection, but the polio vaccine is given by mouth. Many vaccines contain an
first vaccination booster vaccination a few weeks later

booster vaccination at some point in the future

Level of antibody

protective level

4 6 Time / months

Time

Figure 11.14 Antibody levels after vaccination. 234

Chapter 11: Immunology cells that secrete antibodies do not divide. In 1975, a technique was developed to get around this problem (Figure 11.15). B-lymphocytes were fused with cancer cells, which unlike other body cells go on dividing indefinitely. The product of this fusion is called a hybridoma cell. The hybridoma divides repeatedly to form a clone of cells that secrete monoclonal antibodies. When this technique was first invented, noone really knew what uses might be made of it. Since then, many applications have been found for monoclonal antibodies, both in research and in various areas such as medical diagnosis and treatment. Their uses derive from the fact that any particular monoclonal antibody binds very specifically to a particular molecule. blood clots, in the patients body. A very different diagnostic application is in testing for pregnancy. Any couple who are trying for a baby will want to know as soon as possible if the woman has become pregnant. There are now many different pregnancy testing kits on the market which can be used at home. Most of them use monoclonal antibodies to test for the presence of a hormone called human chorionic gonadotrophin (HCG) in her urine. This hormone is only secreted during pregnancy. Monoclonal antibodies are made, using mouse lymphocytes, that will bind specifically with HCG. In one type of pregnancy-testing kit, these HCGspecific antibodies are bound to atoms of gold. The antibodygold complexes are then used to coat the end of a dipstick (Figure 11.16). Another type of monoclonal antibody is also made, which
antigen injection

Using monoclonal antibodies for diagnosis

Monoclonal antibodies can be used to help to diagnose a particular condition, or to find out where particular types of cells are present in the body. Monoclonal antibodies can be used to locate places where blood clots have formed in the body of a person suspected of suffering from deep-vein thrombosis (a blood clot in a vein, often in the leg). First, a mouse is injected with human fibrin, a protein found in blood clots. The fibrin acts as an antigen in the mouse. Mouse B-lymphocytes with the antibody for human fibrin proliferate, especially in the spleen. After a month or so, the spleen contains large quantities of these lymphocytes. The mouse spleen cells are then mixed with cancer cells to form hybridomas, which are checked to see which antibody they secrete. Hybridomas secreting the anti-fibrin antibody are selected and cultured in a fermenter, so that large amounts of the antibody are made. The antibody can be labelled by attaching it to a radioactive chemical that produces gamma radiation. The labelled antibodies are then introduced into the patients blood. As they are carried around the body in the blood stream, they bind to fibrin molecules. A gamma camera can be used to detect the position of the antibodies, an therefore of any

spleen cells mixed and treated to cause cell fusion

cancer cells

hybridoma cells Tiny samples are taken so that only one cell is present in a well. The wells are tested to find a hybridoma cell producing the required antibody.

The hybridoma cell which produces the required antibody is allowed to divide and produce a clone. These cells can be cultured in fermenters, where they will secrete monoclonal antibody.

Figure 11.15 Monoclonal antibody production. 235

Chapter 11: Immunology will specifically bind with HCGantibodygold complexes. These antibodies are impregnated into a region further up the dipstick, called the Patient Test Result region. To use the dipstick, it is dipped into a urine sample. Any HCG in the urine will bind to the antibodies at the end of the stick, which will be carried upwards as the urine seeps up the stick. As the HCGantibodygold complexes reach the test result region of the stick, they bind with the antibodies there and are held firmly in position. As more and more gold atoms arrive there, a pink colour (or another colour, dependent on the
The design of the dipstick

brand) builds up. The stick also contains an area called the Procedural Control Region, which contains yet another type of immobilised monoclonal antibody. These are from goats, and they are anti-mouse antibodies. They bind with the antibodygold complexes even if these have not encountered any HCG in the urine sample. This strip therefore goes pink even if the test result is negative. Other uses are for producing reagents used to determine a persons blood group, the identification and location of some types of cancer and following the progression of an HIV infection.

How the pregnancy dipstick works

1 The stick is dipped into urine to the line. Procedural Control Region (antibodies specific to the antibodies at the end of the stick)

2 HCG-specific antibodies bound to gold are carried up. If there is any HCG present, it binds to the antibodies and is also carried up.

Patient Test Region (immobilised antibody specific to HCG)

antibodies specific to HCG bound to gold

3 If the stick is working, a pink line always appears in the Procedural Control Region HCG-specific antibody bound to gold is carried upwards and captured by antibody specific to it, which was immobilised here.

4 If the urine contains HCG, it binds to the HCG-specific antibody and gold at the end of the dipstick and is carried upwards. When this meets immobilised HCG-specific antibody, it is bound and a pink line appears. This shows the person is pregnant.

Figure 11.16 236

How one type of pregnancy-testing kit works.

Chapter 11: Immunology SAQ 4 Suggest the purpose of the Procedural Control Region on the pregnancy test dip stick. Rituximab is used to treat a cancer called non-Hodgkin lymphoma, in which it is the B-lymphocytes that are the cancerous cells, dividing out of control and producing very large numbers in the body. They form tumours in the lymph nodes. The drug kills both the abnormal (cancerous) B-lymphocytes and also any normal ones, because all of them have CD20 on their surfaces. However, the body continues to produce new B-lymphocytes which are usually normal, rather than cancerous, ones. Rituximab is also used to treat some auto-immune diseases in which overactive B-lymphocytes are implicated, such as rheumatoid arthritis.

Using monoclonal antibodies for treatment

The anticancer drug MabThera is a monoclonal antibody. MabThera is a trade name for the drug rituximab. Rituximab is a monoclonal antibody that binds to a protein called CD20. This protein is found only on the surface of B-lymphocytes. When rituximab binds to these lymphocytes, it destroys them, although the exact mechanism by which it does this is not yet understood.

Summary
bodys immune system responds to the presence of non-self cells or molecules by attacking the The foreign material. This is done by various white blood cells (leucocytes), including phagocytes and lymphocytes. This response is called the immune response. A molecule that initiates an immune response is called an antigen. are mobile cells that are found in almost all parts of the body. They engulf and digest Phagocytes foreign cells or other materials. They include neutrophils and macrophages. Phagocytes often act as antigen-presenting cells, placing antigens from the foreign cells they have engulfed in their plasma membranes, where other cells of the immune system may come into contact with them. are cells that exist in many different varieties. Unlike phagocytes, each individual Lymphocytes lymphocyte is able to respond only to one particular antigen. a B-lymphocyte meets its specific antigen, it responds by dividing to form a clone of When genetically identical plasma cells. These all secrete antibodies that bind to the antigen. a T-lymphocyte meets its specific antigen on the surface of an antigen-presenting cell, it When responds by dividing to form a clone of T-helper cells or T-killer cells. T-helper cells bind to the antigen and secrete cytokines, which stimulate other cells to attack the antigen. T-killer cells also bind to the antigen, and then destroy the cell on which the antigen is present. B-lymphocytes and T-lymphocytes also form clones of memory cells, which remain in the body Both and are able to mount a rapid attack if the same antigen invades the body again. blood plasma contains numerous small protein molecules which together form complement. The When antigens are present in the body, the complement system is activated. The shape of one of the proteins is altered, causing it to become active as an enzyme and remove part of the molecule of another of the complement proteins. This activates the second protein, and so on down the chain, eventually producing large quantities of proteins that help to destroy invading pathogens.
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cells contain granules of substances such as heparin and histamine. When activated, they Mast release their granules and this causes inflammation to occur. The inflammatory response involves the dilation of blood vessels, bringing more lymphocytes and phagocytes to the area. Although mast cells may have a useful role to play, they are involved in inappropriate immune responses to harmless substances, called allergens. They are also involved in some auto-immune diseases. HIV invades a particular type of T-lymphocyte called CD4 cells. This eventually destroys these cells, weakening the immune system and allowing other pathogens to proliferate in the body. are glycoproteins. They are also known as immunoglobulins. Most of them are Y-shaped Antibodies molecules, with binding sites for specific antigens at the tips of the Y. immunity develops when a persons body has responded to the presence of an antigen, and Active has produced a clone of memory cells that can react promptly if the same antigen invades again.
+

This can be achieved through natural exposure to the antigen (natural active immunity) or through vaccination (artificial active immunity). immunity develops when antibodies from elsewhere are introduced into the body. Babies Passive acquire antibodies from their mother through the placenta and in breast milk (natural passive immunity). Antibodies may also be injected into the body (artificial passive immunity). Passive immunity does not last as long as active immunity, because there are no memory cells involved. antibodies are identical antibodies produced from a clone of hybridoma cells. These are Monoclonal produced by fusing a lymphocyte with a cancer cell. Monoclonal antibodies can be used in diagnosis (e.g. in pregnancy tests) or in the treatment of diseases (e.g. MabThera for the treatment of nonHodgkin lymphoma and rheumatoid arthritis).

Questions
Multiple choice questions 1 The following are the steps involved in the process of phagocytosis of a bacterium by a macrophage. I recognition and attachment of bacterium to the phagocyte II attraction of the bacterium and movement of the phagocyte by chemotaxis III intracellular killing and digestion of bacterium IV egestion of epitopes and antigen presentation V fusion of lysosome with a vesicle produced by endocytosis (phagosome) VI engulfment of the bacterium by phagocyte Which of the following shows the correct sequence of the process of phagocytosis? A I II III IV V VI B II III I IV V VI C IV II I III VI V D II I VI V III IV

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2 An immune response is best defined as: A a defensive reaction by the immune system. B a bodily defence reaction which recognises an invading substance and produces a range of cellular and chemical agents directed at the substance. C a reaction which recognises an invading substance. D the bodys reaction to infection. 3 The diagram shows the origin and maturation of B and T-lymphocytes.

bone marrow stem cell

lymphoid precursor cell matures in thymus matures in bone marrow I

II

III

IV

Which of the following correctly identifies cells I, II, III and IV? I II B-cells memory cells T-cells plasma cells III plasma cells B-cells plasma cells B-cells IV memory cells T-cells memory cells T-cells

A B C D

T-cells plasma cells B-cells memory cells

4 Which of the following parts of statements are correct about cell-mediated and humoral immunity? Cell-mediated A B C D involves T-killer cells involves B-cells produces antibodies Humoral involves B-cells involves T-killer cells produces antibodies
continued ...

does not involve cell-to-cell interaction involves cell-to-cell interaction 239

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Why is passive immunity effective for only a short time? A Memory cells are produced. B Antibodies are broken down rapidly. C Antigens enter the body. D Plasma cells are stimulated.

6 Students in a class were exposed to the chicken pox virus by an infected student. What type of immunity would the students obtain if they also became infected? A artificial active immunity B artificial passive immunity C natural active immunity D natural passive immunity 7 8 Which of the following is not true about antibodies? They: A neutralise toxins. B bind to specific antigens. C activate the complement system. D are effective only when attached to the T-cells. Immune responses may be specific or non-specific. Which response is a specific immune response? A capillaries becoming leaky B phagocytosis C release of histamines D production of memory cells

9 Any of the highly specific antibodies produced in large quantity by the clones of a single hybrid cell formed in the laboratory by the fusion of a B cell with a tumour cell are known as: A IgG antibodies. B monoclonal antibodies. C IgM antibodies. D IgA antibodies. 10 Monoclonal antibodies are used in all of the following except: A blood typing for transfusions. B the identification of the location of some types of cancer. C the stimulation of the immune system. D following the progression of HIV infection.

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Structured questions 11 a i What do you understand by the term non-specific immunity? ii Give two examples of non-specific immunity. b The diagram below shows the events occurring during a non-specific response.

[1 mark] [2 marks]

bacteria in cut skin

mast cell

cell 3

produces substance capillary

cell 1 cell 2

i Name the substance produced by the mast cell. ii What are the functions of the substance identified in b i? iii Identify cells 1, 2 and 3. iv Explain the role of the complement system in non-specific immunity. v If the infection lasts for a while the specific immune system is stimulated. What do you understand by the term specific immunity? vi Explain the role of cell 3 in stimulating the specific immune system.

[1 mark] [2 marks] [2 marks] [3 marks] [1 mark] [3 marks]

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12 The diagram below shows the structure of an antibody.

a b c d e f

Name the type of cell that produces antibodies. [1 mark] Copy the diagram above and label the following parts of an antibody: binding site, variable region, constant region, disulphide bonds, light chain, heavy chain, hinge region [3 marks] State one function for each of the following parts: hinge region and disulphide bonds. [2 marks] Explain why a variable region is necessary in the structure of the molecule. [1 mark] Vaccines contain the antigens of pathogens. There are two types of polio vaccine. The Salk vaccine contains dead viral particles while the Sabin vaccine is made of a live attenuated polio virus. The Sabin vaccine replaced the Salk vaccine. i What do you understand by the term antigen? [1 mark] ii State one advantage of using living attenuated viruses to make a vaccine. [2 marks] The graph below shows the concentration of antibody in the blood of a baby after the first oral vaccine and booster shot for polio.
secondary response

Concentration of antibody in blood

primary response

0 first oral vaccine

10

20

30

40

delay

10

20

30

40

second oral vaccine Time / days continued ...

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i Why is there a delay between the time of the first oral vaccine and the first appearance of antibodies in the blood? ii State two ways in which the immune systems primary response differs from the secondary response. iii Explain the differences shown between the primary and secondary responses. 13 a b Distinguish between: i natural and artificial immunity ii active and passive immunity. Copy and complete the table below to indicate the type of immunity attained in each case. Example baby feeding on breast milk child exposed to a friend with chicken pox receiving the MMR vaccine as a child receiving the H1N1 vaccine as an adult getting an emergency tetanus injection after stepping on a rusty nail c d 14 a b c d Describe how an effective vaccine can provide long-term immunity. Explain how passive immunity provides protection to a person who has been bitten by a snake. Describe the mode of action of phagocytes. Define the term immune response. i Describe the origin and maturation of T-lymphocytes. ii Describe the changes that occur to T-lymphocytes during an immune response. i Describe how B-lymphocytes are involved in the immune response. ii Describe the importance of B memory cells in immunity. Type of immunity

[2 marks] [2 marks] [2 marks]

[4 marks]

[5 marks] [4 marks] [2 marks] [3 marks] [2 marks] [2 marks] [3 marks] [3 marks] [2 marks] [1 mark] [4 marks] [2 marks] [2 marks] [2 marks] [4 marks] [3 marks] [7 marks] [5 marks]

Essay questions

15 a i Define the term antibody. ii Make an annotated schematic drawing of an antibody molecule. iii Describe how an antibody acts on bacteria. b i What are monoclonal antibodies? ii Monoclonal antibodies are used for diagnosis and treatment. Identify two examples of each use. iii Describe the use of monoclonal antibodies in pregnancy kits. 16 a b c Distinguish between humoral and cell-mediated immunity. Draw a flow diagram to illustrate the stages of the immune response to an invading pathogen. Explain what is meant by clonal selection and clonal expansion.

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