Molecular Immunology

Objectives
To understand the molecular aspects that
govern body defense mechanisms and the
consequences of their failure
To understand the principles governing
certain immunological techniques used in
clinical practice
Learning outcomes
 By the end of this course, the student is expected to
be able to:
i. Differentiate between innate and acquired
immunity
ii. Explain how the cells of the immune system
develop and are organized
iii. Relate immunoglobulin structure to its functions
iv. Explain how antibody diversity is generated
v. Provide the molecular rationale for
immunoprophylaxis with specific examples
vi. Differentiate between primary and secondary
immunodeficiency
vii. Know the principles governing some
immunological tools
Introduction: Immunology
The branch of biomedicine concerned
with the structure and function of
the immune system, innate and
acquired immunity, the bodily
distinction of self from non-self, and
laboratory techniques involving the
interaction of antigens with specific
antibodies.
Introduction
• The immune system must be able to:
differentiate between material that is a
normal component of the body (“self”) and
material that is not native to the body
“nonself”

• Highly specialized receptors are present for

discriminating between “self” and “nonself”
body components
The Invaders . . .
 Bacteria

 Viruses

 Parasites

such as
fungi,

protista, & worms

Immunity
Innate immunity Adaptive immunity

Humoral Cell-
mediated

Components Components
Macrophages antigen presenting cells
Granulocytes T-cells
Natural killer cells B-cells
Complement Antibodies
Other chemicals: HCL, Complement
lysozyme
Characteristics Characteristics
* Action is immediate * Action requires days to develop
* Response is non-specific * Response is specific
* Response is not enhanced on * Response is enhanced on
repeated exposure to pathogen repeated exposure to pathogen
Overview of the innate immune system
* It is the first line of defense

* It is active at the time of infection

• It consists of:

a- protective cells (WBCs and derivatives)

b- chemical components
The response of the innate immune system
It is divided into two stages:

1- non-inflammatory reactions (body’s static

defenses)
skin, gastric pH, lysozyme in tears, saliva,
mucous

2- local inflammation promotes migration of

phagocytes
and plasma protein into infected tissues
 The phagocytes respond to surface structures present
in large groups of microorganisms (e.g. peptidoglycan,
mannose)
Role of external body surfaces
* The skin consists of sheets of dry, cornified
epithelial cells
 Intact skin act as barrier to bacteria and viruses

* Hair follicles and sebaceous glands produce:

antibacterial substances (fatty acids and
enzymes)

* Normal microbial flora compete with:

potential pathogens
Role of internal body surfaces
The normal movement of fluids and mucous act
as
mechanical factors for cleaning internal
surfaces of:

 Respiratory tract

 Gastrointestinal tract

 Genitourinary tract
Acute Inflammation Resulting From Infection

Inflammation is a nonspecific response of

living tissue to localize and eliminate
the injurious agent.

The injury may be:

physical, chemical or biological
Aquired (specific) immunity
* The acquired immune response is more specialized
than innate immune response

* The acquired immune response involves a combination

of two mechanisms :

1) Humoral immune response

2) cell mediated immune response

* They interact with one another to destroy foreign body

(microorganisms, infected cells, tumor cells)
Aquired (specific) immunity
Two mechanisms
1) Humoral immune response:
- Antibodies are produced by B-lymphocytes
- These have the ability to recognize and bind
specifically to antigen that induced their formation

2) The cell mediated immune response (CMI)

- It is mediated by certain types of T-lymphocytes
- T-lymphocytes recognize foreign material by
means of surface receptors
- T-lymphocytes attack and destroy foreign material
directly or through release of soluble mediators
i.e. cytokines
Characters Of Acquired Immune Response
1) Highly specific for the invading organism

2) Discrimination between “self and “non self” molecules

The response only occurs to “non self” molecules

3) Diversity:
- It can respond to millions of different antigens
- Lymphoctes population consists of many different clones (one
cell and its progeny)
- Each clone express an antigen receptor and responds only to
one antigenic epitope
Mechanism Of Acquired Immune Response
Acquired immune response is initiated by:

* Recognition of the antigen by specific lymphocytes

* Activation of these specific lymphocytes

* Proliferation and differentiation into effector cells;

-The effector cells eliminate the antigen

-Return of homeostasis and development of memory cells

* Memory cells evoke a more rapid and long response on re-

exposure to same antigen
 Types of Acquired Immunity
I. Naturally Acquired Immunity: Obtained in the
course of daily life.
A. Naturally Acquired Active Immunity:
 Antigens or pathogens enter body naturally.
Body generates an immune response to antigens.
Immunity may be lifelong (chickenpox or mumps)
or temporary (influenza or intestinal infections).
B. Naturally Acquired Passive Immunity:
Antibodies pass from mother to fetus via
placenta or breast feeding (colostrum).
No immune response to antigens.
Immunity is usually short-lived (weeks to months).
Protection until child’s immune system develops.
Types of Acquired Immunity
II. Artificially Acquired Immunity: Obtained by receiving a
vaccine or immune serum.

1. Artificially Acquired Active Immunity:

 Antigens are introduced in vaccines (immunization).
 Body generates an immune response to antigens.
 Immunity can be lifelong (oral polio vaccine) or temporary
(tetanus toxoid).

2. Artificially Acquired Passive Immunity:

 Preformed antibodies (antiserum) are introduced into body by
injection.
 Snake antivenom injection from horses or rabbits.
 Immunity is short lived (half life three weeks).
 Host immune system does not respond to antigens.
Other commonly used terms
Serum: Fluid that remains after blood has
clotted and cells have been removed.
Antiserum: Serum containing antibodies to a
specific antigen(s).
 Obtained from injecting an animal (horse, rabbit,
goat) with antigen (snake venom, botulism or
diphtheria toxin).
Serology: The study of reactions between
antibodies and antigens.
Gamma Globulins: Fraction of serum that
contains most of the antibodies.
Duality of Immune System
I. Humoral (Antibody-Mediated) Immunity
 Involves production of antibodies against foreign
antigens.
 Antibodies are produced by a subset of lymphocytes
called B cells.
 B cells that are stimulated will actively secrete antibodies
and are called plasma cells.
 Antibodies are found in extracellular fluids (blood
plasma, lymph, mucus, etc.) and the surface of B cells.
 Provides defense against bacteria, bacterial toxins, and
viruses that circulate freely in body fluids, before they
enter cells.
 Also cause certain reactions against transplanted tissue.
What are antibodies?
They are Proteins that Recognize
Specific Antigens
bind to a particular antigen with very
high specificity
Each antibody has at least two identical
sites that bind antigen: antigen binding
sites.
They belong to a group of serum proteins
called immunoglobulins (Igs).
Antibodies have an antigen-binding site where the epitope of the antigen binds
What is an antigen?
Any organism or molecule, especially proteins or
large polysaccharides that is recognized as foreign
by the body.
Examples:
• Microbes: Capsules, cell walls, toxins, viral capsids,
flagella, etc.
• Nonmicrobes: Pollen, egg white , red blood cell surface
molecules, serum proteins, and surface molecules from
transplanted tissue.
NB: Lipids and nucleic acids are only antigenic
when combined with proteins or polysaccharides.
Antigen…
Generally has molecular weight of 10,000 or
higher.

Hapten
 Is a small foreign molecule that is not antigenic
on its own. It must be coupled to a carrier
molecule to be antigenic.
Once antibodies are formed they will recognize
hapten.
Antigens…
Epitope:
Is a small part of an antigen that
interacts with an antibody.
Any given antigen may have several
epitopes.
Each epitope is recognized by a
different antibody.
Epitopes: Antigen Regions that Interact with
Antibodies
How does the immunoglobulin
structure relate to its functions?
Antibody Structure
 Each antibody (a monomer) has a
flexible Y-shaped molecule with four
polypeptide chains:
 2 identical light chains
 2 identical heavy chains
 Variable Regions:
 Two sections at the end of Y’s arms.

 Contain the antigen binding sites

(Fab).
 Identical on the same antibody, but
vary from one antibody to another.
 Constant Regions:
 They form the stem of monomer
and lower parts of Y arms.
 Fc region: Stem of monomer only.
Important because they can bind to
complement or cells.
Immunoglobulin structure & functional
domains
Ig chains
Light Chain - VL (110 amino acids) and CL
(110 amino acids)
2. Heavy Chain - VH (110 amino acids) and
CH (330-440 amino acids)
There are two types of light chains: kappa (κ)
and lambda (λ)
An Ig molecule has either κκ or λλ but never
κλ
Immunoglobulins: classification
Based on differences in the amino acid
sequences in the constant region of the heavy
chains.
All Igs within a given class have very similar
heavy chain constant regions.
They are:
1. IgG - Gamma heavy chains (γ)
2. IgM - Mu heavy chains (μ)
3. IgA - Alpha heavy chains (α)
4. IgD - Delta heavy chains (δ)
5. IgE - Epsilon heavy chains (ε)
Immunoglobulin Classes IgG
There are four subclasses
Structure: Monomer
Percentage serum antibodies: 80%
Location: Blood, lymph, intestine
Half-life in serum: 23 days
Complement activation: Yes
Placental Transfer: Yes. IgG2 doesn’t cross well.
Known Functions: Enhances phagocytosis,
neutralizes toxins and viruses, protects fetus
and newborn.
II. IgM
 Structure: Pentamer
 First to be made by the fetus
 Percentage serum antibodies:
5-10%
 Location: Blood, lymph, B cell
surface (monomer)
 Half-life in serum: 5 days
 Complement Fixation: Yes
 Placental Transfer: No
 Known Functions: First
antibodies produced during an
infection; effective against lysis
of microbes and agglutinating
antigens.
 III. IgA
 Structure: Dimer.
• Monomers are joined with a J chain.
 Secretory IgA has a secretory component obtained from
intestinal cell surface.
 The secretory piece helps IgA to be transported across mucosa and
also protects it from degradation in the secretions.
 Percentage serum antibodies: 10-15%
 Location: Secretions (tears, saliva, intestine, milk), blood and
lymph.
 Half-life in serum: 6 days
 Complement Fixation: No unless aggregated
 Placental Transfer: No
 Known Functions: Localized protection of mucosal surfaces.
Provides immunity to infant digestive tract.
IV. IgD
Structure: Monomer
Percentage serum antibodies: 0.2%
Location: B-cell surface, blood, and lymph
Half-life in serum: 3 days
Complement Fixation: No
Placental Transfer: No
Known Functions: In serum function is
unknown.
 On B cell surface, initiates immune response.
IgE
Structure: Monomer
Percentage serum antibodies: 0.002%
Location: Bound to mast cells and basophils
throughout the body. Also found in Blood.
Half-life in serum: 2 days
Complement activation: No
Placental Transfer: No
Known Functions: Involved
 in allergic reactions.
 Possibly lysis of worms.
 Protection against parasites
How Do B Cells Produce
Antibodies?
B cells develop from stem cells in the bone
marrow of adults (liver of fetuses).
After maturation, B cells migrate to lymphoid
organs (lymph node or spleen).
Production of antibodies is explained by Clonal
Selection theory
Antibodies are Produced by B Lymphocytes
What does the clonal selection
propose?
The particular kind of antibody to be made is
determined before the cell encounters antigen
Specificity of antibody is determined by its
amino acid sequence.
When a B cell encounters an antigen it
recognizes, it is stimulated and divides into
many clones called plasma cells, which actively
secrete antibodies.
B cell development & activation
Clonal selection theory ...
• Each B cell produces antibodies that will
recognize only one antigenic determinant.
A clone remains after the disappearance of
the antigen and provides immunological
memory.
NB: Immunological tolerance results when a
clone of antigen-binding cells is destroyed or
suppressed.
Tolerance is the absence of the immune
system to respond to antigen
Clonal Selection of B Cells is Caused by Antigenic
Stimulation
Overview of T cell development
Apoptosis
(Programmed cell death)
Human body makes 100 million lymphocytes
every day. If an equivalent number doesn’t
die, leukemia develops.
B cells that do not encounter stimulating
antigen will self-destruct and send signals to
phagocytes to dispose of their remains.
Autoimmunity
Refers to failure of an organism to recognize its
own constituent parts as self, causing an
immune response against its own cells and tissues.
- Any disease that results from such an immune
response is termed an autoimmune disease.
• Autoimmunity is caused by a lack of germ
development of a target body
- the immune response acts against its own cells and
tissues.
- Examples: celiac disease, insulin dependent diabetes
mellitus (Type I DM), systemic lupus erythematosus
(SLE), Hashimoto’s thyroiditis.
Immunological Memory
Antibody Titer: The amount of antibody in the
serum.
Pattern of Antibody Levels During Infection
Primary Response:
 After initial exposure to antigen, no antibodies are
found in serum for several days (lag phase).
 A gradual increase in titer, first of IgM and then of
IgG is observed.
 Most B cells become plasma cells, but some B cells
become long living memory cells.
 Gradual decline of antibodies follows.
Immunological Memory ...
Secondary Response:
Subsequent exposure to the same antigen
displays a faster and more intense antibody
response.
The lag phase is shorter (1-2 days)
Increased antibody response is due to the
existence of memory cells, which rapidly
produce plasma cells upon antigen stimulation.
Antibody Response After Exposure to Antigen
How do antibodies induce resistance?
1) Antitoxin neutralize bacterial toxins (diphtheria,
tetanus)

Antitoxin are developed actively as a result of:

a- Previous infection

b- Artificial immunization

c- Transferred passively as antiserum

* Neutralization of toxin with antitoxin prevents a

combination with tissue cells
How do antibodies induce resistance?...
2) Antibodies attach to the surface of bacteria and

a- act as opsonins and enhance phagocytosis

b- prevent the adherence of microorganisms to

their target cells, e.g. IgA in the gut

c- Activate the complement and lead to bacterial lysis

d- Clump bacteria (agglutination) leading to

phagocytosis
Consequences of Antibody Binding
Consequences of Antigen-Antibody
Binding
Antigen-Antibody Complex: Formed when
an antibody binds to an antigen it recognizes.
May result in the following
1. Agglutination: Antibodies cause
antigens (microbes) to clump together.
 IgM is more effective than IgG
 Hemagglutination: Agglutination of red blood cells
 Used to determine ABO blood types and to detect

influenza and measles viruses.

2. Opsonization: Antigen (microbe) is
covered with antibodies that enhance its
ingestion and lysis by phagocytic cells.
Consequences of Antibody Binding
3. Neutralization: IgG inactivates viruses, drugs, bacteria
by binding to their epitopes and neutralize toxins by
blocking their active sites.
 E.g. diphtheria antitoxin neutralizes the biological effects of
diphtheria toxin.
4. Antibody-dependent cell-mediated cytotoxicity
(ADCC) by IgG, IgE and IgA
 An effector cell of the immune system e.g NK, N, Eu actively
lyse a target cell that has been bound by specific antibodies.
 Used to destroy large organisms (e.g. helminths).
 NK cells release chemicals like cytokines that enter the target
cell and cause cell death.
5. Complement Activation: Both IgG and IgM trigger the
complement system which results in cell lysis and
inflammation.
Phases of humoral immune responses
Vaccination
* Prevents and control diseases e.g. cholera, rabies,
poliomyelitis, diphtheria, tetanus, measles, and
typhoid fever

* Vaccines can be:

a) Prophylactic (i.e. to prevent the effects of
a future infection by any natural or "wild“
pathogen
b) Therapeutic (e.g. vaccines against
cancer
are being investigated)
Immunoprophylaxis: Prevention of infection
through active or passive immunization
 VACCINATIONS
• BIRTH • 9 MONTHS
BCG (BACILLUS CALMETTE-GUERIN) • MEASLES
• ORAL POLIO
• HEPATITIS • 15-18 MONTHS
• MMR (MEASLES, MUMPS, RUBELLA)
• 6 WEEKS
• DPT booster dose
• DPT (DIPHTHERIA, TETANUS,
PERTUSSIS • ORAL POLIO 6TH
• ORAL POLIO 2ND DOSE
• HEPATITIS 2ND • 5 YEARS
• DPT 2ND booster
• 10 WEEKS • ORAL POLIO 7TH
• DPT (DIPHTHERIA, TETANUS,
PERTUSSIS)
• 10 YEARS
• ORAL POLIO 3RD
• TT (TETANUS) 3RD booster
• 14 WEEKS • HEPATITIS B booster
• DPT 3RD
• ORAL POLIO 4TH • 15-16 YEARS
• TETANUS booster
• 6-9 MONTHS
• ORAL POLIO 5TH
• HEPATITIS B
Vaccination
Vaccination:
 Produces immunity against pathogens by the
introduction of:
 live attenuated microorganisms e.g. oral
poliomyelitis (sabine) vaccine, measles, mumps
rubella vaccines, BCG
 killed or inactivated microorganisms e.g.
pertussis and influenza vaccines, or microbial
fractions (e.g. pneumococcal, meningococcal
polysaccharide vaccines)
 altered antigens that stimulate the body to
produce antibodies against more dangerous forms
 Toxoids (e.g. diptheria and tetanus toxoid)
Vaccines work with the immune system's ability
to recognize and destroy antigens.
Passive: Acquisition of preformed antibodies
contained in serum “gamma globulin” or
immunoglobulin fraction(s)
Natural: transplacental passage of martenal IgG
antibodies
Artificial: Administration of antibodies produced
in another host
 (e.g. diphtheria antitoxin; tetanus immune globulin
is Ig fraction of humans hyperimmunized with
tetanus toxoid)
II. Cell Mediated Immunity

Involves T cells that recognize foreign

antigens on the surface of cells, organisms,
or tissues:
 Helper T cells
 Cytotoxic T cells
T cells regulate proliferation and activity of
other cells of the immune system: B cells,
macrophages, neutrophils, etc.
Provide defense against:
 Bacteria and viruses that are inside host cells and
are inaccessible to antibodies.
 Fungi, protozoa, and helminths
 Cancer cells
 Transplanted tissue
Cells Of Immune Response
Cells involved in specific immune mechanisms are:

1) Leucocytes
a) Lymphoid
* T-lymphocytes:
- Antigen specific cells carrying CD3 complex, CD4,
CD8
- Dominant blood lymphocytes (70%)
- Produce cytokines
- Th (T helper) cells: activate and regulate T and B cells
- CD8+Cytotoxic T cells: virus-infected and tumor cells.
- Regulatory (suppressor) T cells: Returns the functioning of
the immune system to normal operation after infection;
prevent autoimmunity
Cells Of Immune Response
* B-lymphocytes:
- Antigen specific cells with surface receptors
- Less common lymphocytes (20%)
- Responsible for antibody production

* NK, K cells:
- Not antigen specific
- Carry Fc receptors , NK-target cell receptor
- Target virus-infected and tumor cells.
Cells of Immune Response….
2-Monocytic myeloid
a- macrophages:
. Non specific
. Carry Fc receptors
. Phagocytic
. Antigen processing and presenting cells
. Produce cytokines
b- Neutrophils:
. Non specific
. Carry Fc receptors, complement
molecules . Mostly target bacteria and fungi
 Cells of Immune Response (cont.)
c- Eosinophils:
. Non specific; mostly target large
parasites
. Carry Fc receptor
. Modulate allergic inflammatory
responses

d- Basophils and Mast cells:

. Non specifc
. Carrying Fc receptors
. Release histamine for allergic
inflammatory responses
Cells of Immune Response…
2-Non hematopoietic cells:
- Dendritic cells
- Function : antigen presenting cells (APC)
that activate T lymphocytes
IMMUNODEFICIENCY
 State in which the immune system's ability to fight
infectious disease is compromised or entirely absent.
 A person with an immunodeficiency is said to be
immunocompromised.
 2 types: primary or secondary.
 Most cases are acquired ("secondary").
1. Primary deficiency:
• One is born with defects in immune system
2. Secondary immunodeficiency
• One is born with normal immune responses but
some secondary factor or occurrence causes a
decrease in immune responses.
Primary or congenital immunodeficiency
Autosomal recessive or X-linked disorders.
Result from an absence or lack of maturation of
T and or B cells.
B cell defects
there is low serum Ig and no seroconversion to
vaccines.
T cell defects
reduced response to skin antigens and reduced
response to viral or fungal infections as well as some
bacterial pathogens e.g. Mycobacteria tuberculosis.
Congenital immunodeficiencies result in
increased infections that are usually manifested
in early infancy and childhood.
Acquired
Common immunodeficiency
causes:
 Malnutrition
 Disseminated cancers
 Treatment with particular drugs e.g.
immunosuppressive drugs, chemotherapy
 Infections of immunocompetent cells with the human
immunodeficiency virus (HIV)
 Aging
Specific diseases that directly or indirectly
impair the immune system
 Some types of cancer:
 bone marrow and blood cells (leukaemia, lymphoma,
multiple myeloma)
 Chronic infections.
 Immunodeficiency is the hallmark of acquired
immunodeficiency syndrome (AIDS)