MID SEM 2003/2004

PART 1
1. Secretory IgA: its production, structures and functions.
Production:
 Tears , saliva, colostrums, mucus
Structure:
 Serum IgA is a monomer but IgA found in secretions is a dimer as presented in Figure below.
 - When IgA exits as a dimer, a J chain is associated with it.
 - When IgA is found in secretions is also has another protein associated with it called the secretory piece or
T piece.
 - sIgA is sometimes referred to as 11S immunoglobulin.
 - Unlike the remainder of the IgA which is made in the plasma cell, the secretory piece is made in epithelial
cells and is added to the IgA as it passes into the secretions (Figure below).
 - The secretory piece helps IgA to be transported across mucosa and also protects it from degradation in
the secretions.
Functions:
 Important in local( mucosal) immunity
2. Explain the principle of the complement fixation test.
Cowey we dnt noe…
3. Answer all:
a. Define the term: Hapten

 Hapten are small molecule which could never induce an immune response and
administered by themselves but which can do when coupled to a carrier molecule.
b. Many phagocytes are cells of the monocyte-macrophage lineage. Mention at least five of them.
 Blood: Monocyte (M. PRECURSOR)
 Tissue : Macrophage (Histocyte)
 Skin: Langerhan cell
 Liver: Kupffer cell
 Bone: Osteoclast
 Lung: Alveolar Macrophages
PART 2
1. What are antigen-presenting cells? Give examples. Discuss the role of the human leucocyte antigens in
antigen presentation.

 APC ARE A HETEROGENOUS POPULATION OF LEUCOCYTES WITH VERY EFFICIENT

IMUNOSTIMULATORY CAPACITY.
 EXAMPLE:
 - B cell
 -Langerhan’s Cell ( skin and other squamous epithelial) interact wth T cell and are termed
interdigitating cells (IDC)
 The follicular Dendritic CELL (FDC) are found in the primary and secondary follicle of the B-CELLS
AREAS OF THE LYMPH NODES AND SPLEEN
 Geminal Centre Dendritic CELL (GCDC) found in germinal centre (GC) of secondary B-cell folicles
 APCs present in thymus are also called IDCs
 Macrophage

2. What are the biological consequences of the activation of the complement system.
Dnt noe… cowey ea..

MID SEM 2004/2005

PART 1
1. Immune response.
There are two types of immune response namely innate immunity and adaptive immunity.
Innate immunity consist of (1) physical and chemicals barrier such as epithelia and antimicrobial
substances produced at epithelial surfaces (2) phagocytic cells (neutrophils, macrophages) and natural killer
(NK) cells (3) blood proteins, including members of the complement system and other mediator of inflammation and
(4) cytokines that regulate the activity of cells of innate immunity.
 There are two types of adaptive immune response i.e. humoral immunity and cell mediated
immunity. Humoral immunity is mediated by antibody. Cell mediated immunity is mediated by T
lymphocytes.
2. Secretory IgA.
1. Structure
- Serum IgA is a monomer but IgA found in secretions is a dimer. When IgA exits as a dimer, a J chain is
associated with it.
- When IgA is found in secretions is also has another protein associated with it called the secretory piece or
T piece.
- sIgA is sometimes referred to as 11S immunoglobulin.
- Unlike the remainder of the IgA which is made in the plasma cell, the secretory piece is made in epithelial
cells and is added to the IgA as it passes into the secretions (Figure below).
- The secretory piece helps IgA to be transported across mucosa and also protects it from degradation in
the secretions.
2. Properties
a) IgA is the 2nd most common serum Ig.
b) IgA is the major class of Ig in secretions - tears, saliva, colostrum, mucus.
- Since it is found in secretions secretory IgA is important in local (mucosal) immunity.
c) Normally IgA does not fix complement, unless aggregated.
d) IgA can binding to some cells - PMN's and some lymphocytes

Ig A: Clinical Implications
Increases in Decreases in

a) Wiskott-Aldrich syndrome a) Hereditary ataxia telangiectasia

b) Cirrhosis of the liver (most cases) b) Immunologic deficiency states (e.g., dysgammaglobulinemia,
congenital and acquired agammaglobulinemia, and
hypogammaglobulinemia)

c) Certain stages of collagen and other autoimmune c) Malabsorption syndromes

disorders such as rheumatoid arthritis and lupus
erythematosus

d) Chronic infections not based on immunologic d) Lymphoid aplasia

deficiencies

e) IgA myeloma e) IgG myeloma

f) Acute lymphoblastic leukemia

g) Chronic lymphoblastic leukemia

 List and briefly describe the steps of phagocytosis mechanism.

STEPS:
1. Margination And Adherence
2. Emigration (Diapedisis)
3. Chemotaxis
4. Engulfment Of The Target
5. Phagosomes Formation
6. Phago-Lysosome Formation
7. Degranulation

1. Margination And Adherence

A. Selectin-mediated phase:
• E-selectin and P-selectin of endocytes bind mucin of leukocytes in less than millisecond .
• This binding is enhanced by microvilli.
B. Activation
• Triggered by leukocyte chemtactic factors:
 Chemokines (produces by endocytes and many other cell).
 Fibrin Collagen.
 Mast cells products (Histamine).
 Complement cascades.
 N-formyl-methionine (Bacteria not in Human proteins).
 Leukocytes chemotactic factors are recognized by specific receptors on leukocytes surface receptors
(Transmembrane).
C. Integrin-mediated phase
• Integrin (Mac-1, LAF-1) bind to CAM-1 on activated endothelium.
2. Emigration (diapedisis)
• This is the migration of leukocytes across vascular walls into adjacent tissue.
3. Chemotaxis
• Phagocytes are motile by amoeboid action.
- This is the movement of cells up a gradient of chemotactic factors.
• It may be directly induced by a substance such as C5a, produced as a result of complement activation.
• It can also be indirectly induced as a consequence of release of preformed mediators within mast cells by
the action of C3a or C5a e.g. eosinophil chemotactic factor, or neutrophil chemotactic factor, Leukotrienes,
produced by the metabolism of mast cell arachidonic acid.
4. Engulfment of the target
• The engulfment of particles (Microorganism, cellular debris, foreign bodies.) is initiated by adherence of a
particle to the surface of the plasma membrane of a phagocyte.
• This step usually involves several types of surface receptors on the phagocyte membrane target recognized
by non-specific surface properties (Hydrophobicity) (un-encapsulated bacteria, Carbon particles) Target
recognized by mannose receptors on the leukocytes (Oligosaccharides).
• Receptor —mediated endocytosis internalizes individual macromolecules on sub-microscopic particles (ex.
Viruses).
• Particles do not interact with any cellular receptors on the cell surface cannot phagocytised directly
(encapsulated bacteria).
• To be phagocytised, encapsulated bacteria need to be coated with certain host derived proteins.
(OPSINISATION)
 important opsinins
o Complement (C3b) mediated through complement receptors on phagocyte surface.
o Immunoglobulin (Ig G) mediated through Fc receptors on phagocyte surface.
5. Phagosomes formation
• The particle is drawn into the cell as the plasma membrane fuses behind the particle, resulting in the
formation, inside the cell, of a membrane-bounded vacuole called a phagosome.
6. Phago-lysosome formation
• The phagosome fuses with a lysosome to form a phagolysosome.
7. Degranulation
• The granules empty their enzymes and other contents into phagolysosome to kill or degrade bacteria or
dissolve other ingested materials.
• The most abundant are the α-defensins (30-50% of total granule proteins.
• Others lysosome contents (antimicrobial) are protease, lactoferrin, transferrin
• Fusion of phagosome with primary granules exposes its content to myeloperoxidase which catalyzes
production of toxic oxidants, halogenation of bacterial proteins and microbial death.
• The contents of phagosome powerful) oxidizing agents generated by (NADPH)-dependent oxidase:
 Singlet oxygen.
 hydrogen peroxide.
 Hypochlorous.
 Organic Chloramine.
3. The principle of the complement fixation test.

PART 2
 Discuss the structure of an immunoglobulin molecule in relation to its functions.
1. Fab - Antigen binding
 These fragments were called the Fab fragments because they contained the antigen binding sites of the
antibody.
 Each Fab fragment is monovalent whereas the original molecule was divalent.
 The combining site of the antibody is created by both VH and VL.
 An antibody is able to bind a particular antigenic determinant because it has a particular combination of VH
and VL.
 Different combinations of a VH and VL result in antibodies that can bind a different antigenic determinants.
2. B. Fc
 This fragment is called Fc (because it is easily crystallized).
 Digestion with papain also produces a fragment that contains the remainder of the two heavy
chains each containing a CH2 and CH3 domain.
 The effector functions of immunoglobulins are mediated by this part of the molecule.
 Different functions are mediated by the different domains in this fragment Normally the ability of
an antibody to carry out an effector function requires the prior binding of an antigen (there are
exceptions to this rule).
3. C. F(ab')2
 Treatment of immunoglobulins with pepsin results in cleavage of the heavy chain after the H-H inter-chain
disulfide bonds resulting in a fragment that contains both antigen binding sites (Figure below).
 This fragment was called F(ab')2 because it was divalent.
 The Fc region of the molecule is digested into small peptides by pepsin.
 The F(ab')2 binds antigen but it does not mediate the effectors functions of antibodies.

1. Bacterial defense against phagocytes.

1. Avoiding Contact with Phagocytes
 Pathogens may remain confined in regions inaccessible to phagocytes.
 Some bacteria or their products inhibit phagocyte chemotaxis (ex. Streptococcal streptolysin, M.
tuberculosis, Clostridium toxin).
 Some pathogens can cover the surface of the bacterial cell with a component which is seen as "self" by the
host phagocytes (ex. Treponema pallidum bind fibronectin to its surface; Streptococci group A synthesize a
capsule composed of hyaluronic acid).
2. Inhibition of Phagocytic Engulfment
 Some bacteria employ strategies to avoid engulfment (ingestion).
 Polysaccharide capsules of S. pneumoniae, Haemophilus influenzae, Treponema pallidum and Klebsiella
pneumoniae.
 M protein and fimbriae of Group A streptococci.
 0 polysaccharide associated with LPS of E. coli.
 K antigen (acidic polysaccharides) of E. col/ or the
o analogous Vi antigen of Salmonella typhi.
 Cell-bound or soluble Protein A produced by
 Staphylococcus aureus.
3. Survival Inside of Phagocytes
 Some bacteria survive inside of phagocytic cells, in either neutrophils or macrophages.
 Bacteria that can resist killing and survive or multiply inside of phagocytes are considered intracellular
parasites.
 Inhibition of fusion of the phagocytic lysosomes (granules) with the phagosome.
 The bacteria survive inside of phagosomes because they prevent the discharge of lysosomal contents into
the phagosome environment..
 Ex. Salmonella, M. tuberculosis, Legionella and the chlamydiae.
4. Products of Bacteria that Kill or Damage Phagocytes
 This is direct attack by the bacteria upon the professional phagocytes.
o A) Killing Phagocytes Before Ingestion
 Many Gram-positive pathogens, particularly the pyogenic cocci, secrete extracellular enzymes that kill
phagocytes.
 Streptococci (streptolysin), Staphylococci (leukocidin), Pseudomonas aeruginosa (Exotoxin A), anthrax
(toxin EF) and pertussis (toxin AC).
 This is a direct attack by the bacteria upon the professional phagocy.
o B) Killing phagocytes after ingestion
 Some bacteria exert their toxic action on the phagocyte after ingestion has taken place.
 o Ex. Mycobacterium, Brucella and Listeria usually destroy macrophages
MID SEM 2007/2008
PART 1
1. Define the following terms:
a. Affinity and avidity
Antibody affinity is the strength of the reaction between a single antigenic determinant and a single
combining site on the antibody. It is the sum of the attractive and repulsive forces operating between the antigenic
determinant and the combining site of the antibody. Affinity is the equilibrium constant that describes the Ag-Ab
reaction.
Avidity is a measure of the overall strength of binding of an antigen with many antigenic determinants and
multivalent antibodies.
b.Specificity and cross reactivity
Specificity refers to the ability of an individual antibody combining site to react with only one antigenic
determinant or the ability of a population of antibody molecules to react with only one antigen. In general, there is a
high degree of specificity in Ag-Ab reactions.
Cross reactivity refers to the ability of an individual antibody combining site to react with more than one
antigenic determinant or the ability of a population of antibody molecules to react with more than one antigen.
c.Immunogen
A substance tt induce a specific immune response.
d.Epitope
-also known as antigenic determinant
- D portion of an Ag tt combine with the products of a specific immune response.
e. Hapten
-A small molecule tt is non-immunogenic
-Hv antigenicity property but not immunogenicity
-Can react with the product specific immune response.
2. Positive and negative selection of developing T cells.
Positive and negative selection of developing T cells occurs in the thymus
1. Positive selection
• T cells recognize antigenic peptides only when presented by self-MHC molecules on APCs.
• T cells show `dual recognition' of both the antigenic peptides and the polymorphic part of the MHC
molecules.
• Positive selection (also called thymic education) ensures that only those TCRs with a moderate affinity for
self MHC are allowed to develop further.
• T cells displaying very high or very low receptor affinities for self MHC undergo apoptosis and die in the
cortex.
2. Negative selection
• Some of the positively selected T cells may have receptors that recognize self components other than self
MHC.
• These cells are deleted by a `negative selection' process.
• Not all self-reactive T cells are eliminated during intrathymic development.
• This probably occurs because not all self antigens are able to be presented in the thymic tissues.
• Thus, negative selection may also occur in the periphery.
3.Immunoglobulin fragments produced by proteolytic digestion have been proven very useful in elucidating structure-
function relationships in immunoglobulins. Discuss the fragments digested with papain.
a) -Digestion with papain breaks an Ig molecule into d hinge region before d H-H interchain disulfide bond.
-Results d formation of 2 identical fragments tt contain d light chain and d VH and CH1 domains of d
heavy chain.
-these fragments are called Fab fragments cz they contain d Ag binding site of d Ab. Each fragment is
monovalent whereas d original is divalent.
b) –Digestion with papain also produced a fragment tt contain d remainder of d 2 heavy chains
each containing a CH2 and CH3 domain.
-D fragments r called Fc (mediate d effectors function)
Key words:
D=the
Tt=that
Hv=have
IMMUNOLOGY 2007/2008
 1) Factors influencing immunogenicity.
2) Bacterial defense against phagocytes.
Answers:
1) Factors influencing immunogenicity:
a) Contribution of the immunogen
i. Foreigness – the immune system usually discriminates between self and non-self such that only foreign
molecules are immunogenic.
ii. Size – there is no absolute size above which a substance will be immunogenic. However, generally, the
larger the molecule the more immunogenic it is likely to be.
iii. Chemical composition – in general, the more complex the substance is chemically, the more immunogenic it
will be. The antigenic determinants are created by the primary sequence of residues in the polymer and/or
by the secondary, tertiary or quaternary structure of the molecule.
iv. Physical form – in general, particulate antigens are more immunogenic than soluble ones and denatured
antigens more immunogenic than the native form.
v. Degradability – antigens that are easily phagocytosed are generally more immunogenic. This is because for
most antigens (T-dependant antigens) the development of an immune response requires that the antigen
be phagocytosed, processed and presented to helper T cells by and antigen presenting cell (APC).
b) Contribution of the biological system
i. Genetic factors – some substances are immunogenic in one species, one individual, but not in another. The
species or individuals may lack or have altered genes that code the receptors for antigen on B cells and T
cells or they may not have the appropriate genes needed for the APC to present antigen to the helper T
cells.
ii. Age – usually the very young and the very old have a diminished ability to mount and immune response in
response to an immunogen.
c) Method of administration of an immunogen
i. Dose – there is a dose of antigen above or below which the immune response will not be optimal.
ii. Route – the route of antigen administration can also alter the nature of the response. Generally, the
subcutaneous route is better that the intravenous or intragastric routes.
iii. Adjuvants – they are the substances that can enhance the immune response to an immunogen. The use of
adjuvants is often accompanied with undesirable side effects such as fever and inflammation.
2) Bacterial defense against phagocytes:
i. Avoiding contact with phagocytes
- Pathogens may remain confined in regions inaccessible to phagocytes. Some bacteria or their products
inhibit phagocyte chemotaxis. Some pathogens can cover the surface of the bacterial cell with a component
which is seen as ‘self’ by the host phagocytes.
ii. Inhibition of phagocytic engulfment
- Some bacteria employ strategies to avoid engulfment:-
 Polysaccharide capsules
 M protein and fimbriae
 O polysaccharide
 K antigen
 Cell-bound or soluble Protein A
iii. Survival inside of phagocytes
- Bacteria that can resist killing and survive or multiply inside of phagocytes are considered intracellular
parasites.
- Inhibition of fusion of the phagocytic lysosomes (granules) with the phagosome. The bacteria survive inside
of phagosomes because they prevent the discharge of lysosomal contents into the phagosome
environment.
iv. Products of bacteria that kill or damage phagocytes
- This is the direct attack by the bacteria upon the professional phagocytes.
 Killing phagocytes before ingestion – many gram positive pathogens secrete extracellular enzymes
that kill phagocytes.
 Killing phagocytes after ingestion – some bacteria exert their toxic action on the phagocyte after
ingestion has taken place.

END SEM 2004/2005

PART 1
Answer three (3) out of the following:
(Marks = 5% per question)
1. Briefly discuss the differences between the two types of immunity (specific or adaptice immunity and non-
specific immunity).

Non-specific immunity Specific Immunity

Response is antigen-independent Response is antigen-dependent

There is a lag time between exposure and maximal

There is immediate maximal response
response

Not antigen-specific Antigen-specific

Ability to recognize, mainly, carbohydrates, lipids

Recognizes pathogens mainly by specific proteins
and nucleic acids

Exposure results in non-immunologic memory Exposure results in immunologic memory

2. In brief discuss mechanisms that have been proposed for the escape of malignant cells from host immune-
surveillance.
Several mechanisms have been proposed for the escape of malignant cells from host immuno-
surveillance:
1. tumours may not express neo-antigens that are immunogenic or
2. They may fail to express co-stimulatory molecules for the activation of T-cells.
3. Certain tumours have been known to lack or be poor in expression of MHC antigens.
4. In the early development of a tumour, the amount of antigen may be too small to stimulate the
immune system
5. due to the rapid proliferation of malignant cells, the immune system is quickly overwhelmed.
6. some tumours may avoid the immune system by secreting immunosuppressive molecules and
others by inducing suppressor cells.
7. some tumours may shed their unique antigens which block antibodies and T cells from reacting
with malignant cells.
3. Mention the biological functions of the complement system.
Biologically active products of Complement activation
- Activation of complement results in the production of several biologically active molecules which contribute
to
- resistance
- anaphylaxis
- inflammation
a) Kinin production
- C2b generated during the classical pathway of C activation is a prokinin which becomes biologically active
following enzymatic alteration by plasmin
Excess C2b production is prevented by limiting C2 activation by C1 inhibitor (C1-INH) also known as serpin
which displaces C1rs from the C1qrs complex
- A genetic deficiency of C1-INH results in an overproduction of C2b and is the cause of hereditary
angioneurotic edema.
- This condition can be treated with Danazol which promotes C1-INH production or with e-amino caproic acid
which decreases plasmin activity.
b) Anaphylotoxins:
- C4a, C3a and C5a (in increasing order of activity) are all Anaphylotoxins which cause basophil/mast cell
degranulation and smooth muscle contraction.
- Undesirable effects of these peptides are controlled by carboxypeptidase B (C3a-INA).
 c) Chemotactic Factors:
- C5a and MAC (C5b67) are both chemotactic
- C5a is also a potent activator of neutrophils, basophils and macrophages.
- C5a causes induction of adhesion molecules on vascular endothelial cells.
d) Opsonins:
- C3b and C4b in the surface of microorganisms attach to C-receptor (CR1) on phagocytic cells
and promote phagocytosis.
4. Mention the mechanisms of breakdown of immunological tolerance to self antigens.
i. Sequestered antigen
• Lymphoid cells may not be exposed to some self antigens during their differentiation, because they may be
late-developing antigens or may be confined to specialized organs (e.g., testes, brain, eye, etc.).
A release of antigens from these organs resulting from accidental traumatic injury or surgery can result in
the stimulation of an immune response and initiation of an autoimmune disease.
ii. Escape of auto-reactive clones
• The negative selection in the thymus may not be fully functional to eliminate self reactive cells.
• Not all self antigens may be represented in the thymus or certain antigens may not be properly processed
and presented.
iii. Cross reactive antigens
• Antigens on certain pathogens may have determinants which cross react with self antigens and an immune
response against these determinants may lead to effector cell or antibodies against tissue antigens.
• Post streptococcal nephritis and carditis, anticardiolipin antibodies during syphilis and association between
Klebsiella and ankylosing spondylitis are examples of such cross reactivity.