Basic Immunology and Vaccines

Chandani Udawatte
• Immunology is the study of the structure and
function of the immune system.

• Vaccinology is the science of vaccine development

and how the immune system responds to vaccines,
but also includes ongoing evaluation of
immunization programs and vaccine safety and
effectiveness, as well as surveillance of the
epidemiology of vaccine-preventable diseases.
 Human Immune System
Components of the immune system
An antigen is a substance that the body recognizes as foreign and that triggers
immune responses.
The terms immunogen and antigen are often used interchangeably.

Antibodies are proteins that are produced in response to antigens introduced

into the body.

Antibodies protect the body from disease by,

• binding to the surface of the antigen to block its biological activity
(neutralization)
• binding or coating (opsonisation) of the antigen to make it more susceptible
to destruction and clearance by phagocytes (phagocytosis)
• opsonisation of special receptors on various cells, allowing them to
recognise and respond to the antigen
• activation of the complement system to cause disintegration (lysis) of the
pathogen and to enhance phagocytosis.
 Immune responses
• Immunity is the ability of the human body to
protect itself from infectious diseases. The
human immune system is able to react to an
enormous number and variety of foreign
antigens and provides immunity through two
complementary types of responses, innate
immunity and adaptive immunity.
 Innate immunity
• Innate immunity is the body’s initial defence mechanism that is
not specific to particular antigens and comes into play
immediately or within hours of a pathogen’s entry into the body.

• Innate immunity is made up of physical barriers (skin and mucous

membranes), physiologic defences (temperature, low pH and
chemical mediators), and the human microbiome (aggregate of
microorganisms that reside on and in the human body; these
microorganisms may protect from potential pathogens by blocking
binding sites and competing for nutrients), as well as phagocytic
and humoral inflammatory responses.

• Innate immunity,
– does not depend upon previous exposure to the pathogen
– does not produce immunologic memory
– does not improve with repeated exposure to the pathogen.
 Adaptive immunity
• Adaptive immunity is the body’s second level of defence,
which develops as a result of infection with a pathogen or
following immunization. It defends against a specific
pathogen and takes several days to become protective.

• Adaptive immunity,
– has the capacity for immunologic memory
– provides long-term immunity which may persist for a lifetime
but may wane over time
– increases in strength and effectiveness each time it encounters
a specific pathogen or antigen.

• The cells of the adaptive immune system include specialized

white blood cells:
T cells (T lymphocytes) - cell-mediated immunity and
B cells (B lymphocytes) - antibody-mediated immunity
• Cell-mediated immunity provides protection through
activation of specialized T cells to destroy pathogens or to
induce the death of cells displaying the foreign antigen on
their surface, and stimulation of further immune response.

• Antibody-mediated (humoral) immunity is mediated by B

cells. The terms antibody and immunoglobulin (Ig) are often
used interchangeably. There are five types of antibodies: IgA,
IgD, IgE, IgG and IgM.

• Immunologic memory is the immune system’s ability to

remember its experience with an infectious agent, leading to
effective and rapid immune response upon subsequent
exposure to the same or similar infectious agents.
Development of immunologic memory requires participation
of both B and T cells; memory B cell development is
dependent on the presentation of antigens by T cells.
 Immunization & Immunizing
Agents
• Immunization refers to the process by which a person becomes protected
against a disease with immunizing agents.

• Immunizing agents are classified as active or passive, depending on the

process by which they confer immunity; prevention of disease through the
use of immunizing agents is called immunoprophylaxis.

• Active immunization is the production of antibodies against a specific agent

after exposure to the antigen through vaccination. Active immunizing
agents are typically referred to as vaccines.

• Passive immunization involves the transfer of pre-formed antibodies,

generally from one person to another or from an animal product, to provide
temporary protection, since transferred antibody degrades over time. It can
occur by transplacental transfer of maternal antibodies to the developing
foetus, or it can be provided by administration of a passive immunizing
agent prepared from the serum of immune individuals or animals.
 Active vaccines
• Vaccines are complex biologic products designed to induce a protective immune response
effectively and safely.
• An ideal vaccine is safe with minimal adverse effects, and effective in providing lifelong
protection against disease after a single dose that can be administered at birth. Also ideally,
it would be inexpensive, stable during shipment and storage, and easy to administer. Some
vaccines come closer to fulfilling these criteria than others. Although each vaccine has its
own benefits and risks, and indications and contraindications, all vaccines offer protection
against the disease for which they were created.

• In addition to the active component, the antigen, which induces the immune response,
vaccines may contain additional ingredients such as preservatives, additives, adjuvants and
traces of other substances necessary in the production of the vaccine.

• Vaccine antigens include inactivated (killed) or attenuated (weakened) live organisms,

products secreted by organisms that are modified to remove their pathogenic effects (e.g.,
tetanus toxoid), and components of the organism - some of which are made in the
laboratory through recombinant technology.
 Classification of vaccines
• Vaccines are classified according to the type of antigen they contain.

• Most often they are categorized in two groups - live attenuated vaccines
and inactivated vaccines

• Inactivated vaccines contain whole or part of (fractional) killed bacteria or

viruses and cannot cause the disease it is designed to prevent, even in an
immunocompromised person. Fractional inactivated vaccines can be
protein or polysaccharide-based. Antigens of protein-based vaccines
include toxoids (inactivated bacterial toxin), subunit and split-virus products.
– Subunit vaccines are highly purified products containing surface
antigen only, with most (if not all), of the internal viral or bacterial
components removed. Some subunit vaccines are synthesized using
recombinant technology (e.g. hepatitis B vaccine). Subunit vaccines
have excellent safety profiles and are used in a variety of
combination products.
– Split-virus vaccines are treated to disrupt the integrity of the virus
but contain essentially all viral structural proteins and components.
 Live attenuated vaccines
• Live attenuated vaccines contain whole, weakened bacteria or viruses.

• Since the agent replicates within the vaccine recipient, the stimulus to the
immune system more closely resembles that associated with natural
infection, resulting in longer lasting and broader immunity than can be
achieved with other vaccine types.

• Because of the strong immunogenic response, live attenuated vaccines,

except those administered orally, typically produce immunity in most
recipients with one dose; however, a second dose helps to make sure that
almost everyone is protected, because some individuals may not respond to
the first dose.

• Live vaccines require careful storage and handling to avoid inadvertent

inactivation and are, in general, contraindicated for pregnant women,
because of the risk of effects on the foetus, and for people who are
immunocompromised, due to the risk of disease being caused by the live
vaccine strains.
 Inactivated vaccines
• Polysaccharide vaccines are composed of bacterial capsule and are
less immunogenic compared to other vaccines, particularly in
children younger than two years of age. However, polysaccharides
may be chemically joined or linked to a carrier protein (ie. a
protein that is easily recognized by the immune system, ie.
diphtheria or tetanus, to produce conjugate polysaccharide
vaccines that have improved functional activity and are highly
immunogenic in young children.

• Because the immune response to inactivated vaccines may be less

than that induced by live organisms, inactivated vaccines often
require multiple doses: the first dose primes the immune system
and a protective immune response develops after the second or
third dose. These initial doses are called primary vaccination orthe
primary series. Because protection following primary vaccination
may diminish over time, periodic supplemental doses, or booster
doses, may be required to increase or boost antibody levels.
 Live attenuated vaccines and inactivated
vaccines
 Live attenuated vaccines
Live attenuated bacterial vaccines
BCG vaccine
Typhoid vaccine (oral formulation)

Live attenuated viral vaccines

Live attenuated influenza vaccine (intranasal formulation)
Measles-mumps-rubella containing vaccines
Rotavirus vaccines
Varicella-containing vaccines, including herpes zoster vaccine
Yellow fever vaccine
 Inactivated vaccines
Whole inactivated vaccines
Viruses (e.g., polio, hepatitis A, rabies vaccines)
Bacteria (e.g., cholera vaccine)

Fractional inactivated vaccines

•Protein-based
Toxoid (e.g., diphtheria toxoid, tetanus toxoid)
Subunit (e.g., hepatitis B, acellular pertussis, some influenza vaccines)
Split-virus (e.g., some influenza vaccines)
•Polysaccharide-based
Pure polysaccharide (e.g., pneumococcal polysaccharide, parenteral
typhoid vaccine)
Conjugate polysaccharide (e.g., pneumococcal conjugate, meningococcal
conjugate, Haemophilus influenzae type b [Hib] vaccines)
•Virus-like particle (e.g., human papillomavirus [HPV])
 Passive immunizing agents
• Passive immunization with immune globulins provides protection when
vaccines for active immunization are unavailable or contraindicated, or in
certain instances when unimmunized individuals have been exposed to the
infectious agent and rapid protection is required (post-exposure
immunoprophylaxis).

• Passive immunization also has a role in the management of

immunocompromised people who may not be able to respond fully to
vaccines or for whom live vaccines may be contraindicated. The duration of
the beneficial effects provided by passive immunizing agents is relatively
short and protection may be incomplete.

• Passive immunizing agents are preparations containing pre-formed

antibodies derived from humans or animals, or produced by recombinant
DNA technology. Administration of passive immunizing agents can prevent
certain infections or reduce the severity of illness caused by the infectious
agent.
 Antibody preparations
There are two types of antibody preparations available:

• Standard immune globulin (Ig) of human origin

Sometimes referred to as “immune serum globulin”, “serum
immune globulin” or “gamma globulin”

• Specific immune globulins of human or animal origin, or produced

by recombinant DNA technology
Contains high titres of specific antibodies against a particular
microorganism or its toxin.
Products of human origin are preferred over those of animal origin
because of the high incidence of adverse reactions to animal sera
and the longer lasting protection conferred by human immune
globulins.
 Standard Immune globulin (human)
• Standard human immune globulin (GamaSTAN® S/D) is a sterile,
concentrated solution for intramuscular (IM) injection containing
15% to 18% immune globulin.
It is obtained from pooled human plasma from screened donors
and contains mainly IgG with small amounts of IgA and IgM.
The potency of each lot of immune globulin product is tested
against international standards or reference preparations for at
least two different antibodies, one viral and one bacterial.

• Subcutaneous (Sc) and intravenous (IV) immune globulin (IVIg) are

used for continuous passive immunization for persons with selected
congenital or acquired immunoglobulin deficiency states and as an
immunomodulator in certain diseases.
 Specific immune globulins
• Specific immune globulins are derived from
the pooled sera of people with antibodies to specific infectious agents;
antisera from animals (usually horses that are hyper-immunized against a
specific organism) when human products are not available;
or recombinant DNA technology

• Immune globulins from human or animal sources are made by more than
one B cell clone (polyclonal) and can bind to heterogeneous antigens.

Antibodies produced through recombinant DNA technology originate from

a single clone of B cells (monoclonal) and are specific to only one antigen;
e.g. a monoclonal antibody product is available for the prevention of
respiratory syncytial virus (RSV) infection.

Because of the relatively high risk of a specific type of immunological

reaction (known as serum sickness) following the use of animal products,
human Ig should be used whenever possible.
 Immunoglobulin (Ig)

• Immunoglobulins are glycoprotein molecules that are

produced by plasma cells in response to an
immunogen and which function as antibodies. The
immunoglobulins derive their name from the finding
that they migrate with globular proteins when
antibody-containing serum is placed in an electrical
field
 GENERAL FUNCTIONS OF
IMMUNOGLOBULINS
• Antigen binding

Immunoglobulins bind specifically to one or a few closely

related antigens. Each immunoglobulin actually binds to a
specific antigenic determinant. Antigen binding by antibodies
is the primary function of antibodies and can result in
protection of the host. The valency of antibody refers to the
number of antigenic determinants that an individual antibody
molecule can bind. The valency of all antibodies is at least two
and in some instances more.
• Effector Functions
Frequently the binding of an antibody to an antigen has no direct
biological effect. Rather, the significant biological effects are a
consequence of secondary "effector functions" of antibodies. The
immunoglobulins mediate a variety of these effector functions.
Usually the ability to carry out a particular effector function
requires that the antibody bind to its antigen.

• Not every immunoglobulin will mediate all effector functions. Such

effector functions include:
1. Fixation of complement - This results in lysis of cells and release
of biologically active molecules
2. Binding to various cell types - Phagocytic cells, lymphocytes,
platelets, mast cells, and basophils have receptors that bind
immunoglobulins. This binding can activate the cells to perform
some function. Some immunoglobulins also bind to receptors on
placental trophoblasts, which results in transfer of the
immunoglobulin across the placenta. As a result, the transferred
maternal antibodies provide immunity to the fetus and newborn
 BASIC STRUCTURE OF
IMMUNOGLOBULINS
All immunoglobulins have the same basic units, although different
immunoglobulins can differ structurally

• Heavy and Light Chains

All immunoglobulins have a four chain structure as their basic unit.
They are composed of two identical light chains (23kD) and two
identical heavy chains (50-70kD)

• Disulfide bonds
Inter-chain disulfide bonds - The heavy and light chains and the
two heavy chains are held together by inter-chain disulfide bonds
and by non-covalent interactions. The number of inter-chain
disulfide bonds varies among different immunoglobulin molecules.
• Intra-chain disulfide binds - Within each of the polypeptide chains
there are also intra-chain disulfide bonds.
• Variable (V) and Constant (C) Regions
When the amino acid sequences of many different heavy
chains and light chains were compared, it became clear
that both the heavy and light chain could be divided into
two regions based on variability in the amino acid
sequences. These are the:
Light Chain - VL (110 amino acids) and CL (110 amino acids)
Heavy Chain - VH (110 amino acids) and CH (330-440 amino
acids)

• Hinge Region
This is the region at which the arms of the antibody
molecule forms a Y. It is called the hinge region because
there is some flexibility in the molecule at this point.
• Domains
Three dimensional images of the immunoglobulin
molecule show that it is not straight Rather, it is folded
into globular regions each of which contains an intra-
chain disulfide bond. These regions are called
domains.
Light Chain Domains - VL and CL
Heavy Chain Domains - VH, CH1 - CH3 (or CH4)

• Oligosaccharides
Carbohydrates are attached to the CH2 domain in most
immunoglobulins. However, in some cases
carbohydrates may also be attached to other locations.
 IMMUNOGLOBULIN FRAGMENTS: STRUCTURE/FUNCTION
RELATIONSHIPS
Immunoglobulin fragments produced by proteolytic digestion have proven very useful
in elucidating structure/function relationships.

• Fab
Digestion with papain breaks the immunoglobulin molecule in the hinge region
before the H-H inter-chain disulfide bond. This results in the formation of two
identical fragments that contain the light chain and the VH and CH1 domains of the
heavy chain.

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.
• Fc
Digestion with papain also produces a fragment that
contains the remainder of the two heavy chains each
containing a CH2 and CH3 domain. This fragment was called
Fc because it was easily crystallized.

• 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 . This fragment was called
F(ab')2because it is divalent. The Fc region of the molecule
is digested into small peptides by pepsin. The F(ab')2binds
antigen but it does not mediate the effector functions of
antibodies.
 Immunoglobulin Classes
• The immunoglobulins can be divided into five different
classes, based on differences in the amino acid sequences
in the constant region of the heavy chains.

All immunoglobulins within a given class will have very

similar heavy chain constant regions. These differences can
be detected by sequence studies or more commonly by
serological means (i.e. by the use of antibodies directed to
these differences).

• IgG - Gamma heavy chains

• IgM - Mu heavy chains
• IgA - Alpha heavy chains
• IgD - Delta heavy chains
• IgE - Epsilon heavy chains
 Immunoglobulin Subclasses
• The classes of immunoglobulins can de divided into subclasses based on
small differences in the amino acid sequences in the constant region of the
heavy chains. All immunoglobulins within a subclass will have very similar
heavy chain constant region amino acid sequences. Again these differences
are most commonly detected by serological means.

• 1. IgG Subclasses
a) IgG1 - Gamma 1 heavy chains
b) IgG2 - Gamma 2 heavy chains
c) IgG3 - Gamma 3 heavy chains
d) IgG4 - Gamma 4 heavy chains

• 2. IgA Subclasses
a) IgA1 - Alpha 1 heavy chains
b) IgA2 - Alpha 2 heavy chains
 Immunoglobulin Types –
type of light chain
• Immunoglobulins can also be classified by
the type of light chain that they have. Light
chain types are based on differences in the
amino acid sequence in the constant region of
the light chain. These differences are detected
by serological means.

• 1. Kappa light chains

• 2. Lambda light chains
 Immunoglobulin Subtypes – type of
light chain
•
The light chains can also be divided into subtypes
based on differences in the amino acid sequences
in the constant region of the light chain.

• 1. Lambda subtypes
a) Lambda 1
b) Lambda 2
c) Lambda 3
d) Lambda 4
• Nomenclature
Immunoglobulins are named based on the class, or
subclass of the heavy chain and type or subtype of light
chain. Unless it is stated precisely, you should assume
that all subclass, types and subtypes are present. IgG
means that all subclasses and types are present.

• Heterogeneity
Immunoglobulins considered as a population of
molecules are normally very heterogeneous because
they are composed of different classes and subclasses
each of which has different types and subtypes of light
chains. In addition, different immunoglobulin
molecules can have different antigen binding
properties because of different VH and VL regions.
 Adjuvants
• An adjuvant is a substance that is added to a vaccine to enhance
the immune response and to extend the duration of B and T cell
activation.

• An adjuvant allows the reduction of the amount of antigen per

dose or the total number of doses needed to achieve immunity
and helps to improve the immune response in individuals with
some degree of immune suppression (e.g., the elderly).

• Some of the adjuvants used in vaccines currently marketed are,

aluminum salts (aluminum hydroxide, aluminum phosphate, or
aluminum hydroxyphosphate sulfate)
AS04 (3-O-desacyl-4’-monophosphoryl lipid A adsorbed onto
aluminum [as hydroxide salt])
MF59C.1 (oil-in-water emulsion composed of squalene as the oil
phase, stabilised with the surfactants polysorbate 80 and sorbitan
trioleate, in citrate buffer).
 Preservatives
• Chemicals such as thimerosal, phenol,
2-phenoxyethanol, may be added to vaccines
to prevent serious secondary infections as a
result of bacterial or fungal contamination of
the vaccine.
 Additives
Additives are substances that may be added to vaccines to,
• Support the growth or purification of specific antigens or the
inactivation of toxins, or both
They include antibiotics added to prevent contamination
during viral cell culture
substances needed for the growth of viruses such as egg or
yeast proteins, glycerol, serum, amino acids and enzymes
formaldehyde used to inactivate viruses and protein toxins
Most of these reagents are removed in subsequent
manufacturing steps, but minute amounts may remain in the
final product.
• Support product quality or stability
Compounds may be added to vaccines for a variety of
manufacture-related issues
controlling acidity (pH)
stabilizing antigens through the manufacturing process,
such as during freeze drying (lyophilizing)
preventing antigens from adhering to the sides of glass
vials with a resultant loss in immunogenicity
Examples of such additives include potassium or
sodium salts, lactose, polysorbate 20 or 80, human
serum albumin and animal proteins such as gelatin and
bovine serum albumin
 How Vaccines Work

• Vaccines work at an individual level to protect

the immunized person against the specific
disease, as well as at a population level to
reduce the incidence of the disease in the
population, thereby reducing exposure of
susceptible persons and consequent illness.
 How vaccines work at the individual
level
• The administration of a vaccine antigen triggers an
inflammatory reaction that is initially mediated by the
innate immune system and subsequently expands to
involve the adaptive immune system through the activation
of T and B cells.

• While the majority of vaccines provide protection through

the induction of humoral immunity, primarily through B
cells, some vaccines such as Bacille Calmette-Guerin (BCG)
and herpes zoster act principally by inducing cell-mediated
immunity primarily though T cells.

• Long-term immunity requires the persistence of antibodies,

and/or the creation and maintenance of antigen-specific
memory cells (priming), that can rapidly reactivate to
produce an effective immune response upon subsequent
exposure to the same or similar antigen.
• Immunogenicity means the vaccine’s ability to
induce an immune response.

• Vaccine-induced seroconversion is the

development of detectable antigen-specific
antibodies in the serum as a result of vaccination

• Seroprotection is a predetermined antibody level

as a result of vaccination, above which the
probability of infection is low. The seroprotective
antibody level differs depending on the vaccine.
 Vaccines at the population level
• Vaccine efficacy refers to the vaccine’s ability to prevent illness in people
vaccinated in controlled studies.
• Vaccine effectiveness refers to the vaccine’s ability to prevent illness in
people vaccinated
• Herd immunity
Herd immunity refers to the immunity of a population against a specific
infectious disease. The resistance of that population to the spread of an
infectious disease is based on the percentage of people who are immune
and the probability that those who are still susceptible will come into
contact with an infected person.

The proportion of the population required to be immune to reach herd

immunity depends on a number of factors, the most important one being
the transmissibility of the infectious agent either from a symptomatically
infected person or from an asymptomatically colonized person.
The reproduction number (also called the basic reproductive
rate) or R0 is defined as the average number of
transmissions expected from a single primary case
introduced into a totally susceptible population. Diseases
that are highly infectious have a high R0 (e.g., measles) and
require higher vaccine coverage to attain herd immunity
than a disease with a lower R0 (e.g., rubella, Hib). To stop
transmission of a given disease, there needs to be at least a
specified percentage (1 minus 1/R0) of the population
immune to the disease.
For example, measles has an estimated R0 of 15; therefore,
at least 94% (1 minus 1/15 = 94%) of the population needs
to be immune to prevent transmission of measles.
 Vaccine Development
The first steps in the development of a vaccine include the identification
of the microorganism or toxin that causes a significant burden of
disease in the population and an understanding of the disease
pathogenesis.

Once the pathogen and pathogenesis are understood, research is

initiated into the possibility of developing a vaccine to reduce the
disease incidence, severity or both.

New vaccines undergo a very rigorous development process, beginning

with pre-clinical laboratory testing to ensure that the candidate vaccine
produces the immune response needed to prevent disease and has no
toxicities that would prevent its use in people.

Clinical trials on humans then proceed through several phases involving

progressively more study subjects.
 Epidemiology and Immunization
• Epidemiology provides data on the distribution and determinants
of diseases. As a vaccine is introduced into the population,
epidemiology monitors the effect of the vaccine in the population
by describing changes in the disease burden and the pathogens
causing that disease. Epidemiology can also provide information
regarding immunization coverage and vaccine safety.

• Surveillance is the process of systematic collection, orderly analysis,

evaluation and reporting of epidemiological data, particularly to
public health officials who are in a position to take action, to inform
disease control measures or policy decisions or both. Surveillance
of vaccine preventable diseases, including immunization coverage
and vaccine safety, is needed to identify and quantify risk factors to
enable appropriate control of communicable diseases, assist in the
management of vaccine preventable disease outbreaks and signal
adverse events following immunization, monitor progress toward
the achievement of set goals and targets in disease control
programmes and provide up-to-date information to develop
evidence-based guidelines.
 Future of vaccinology

Improving the effectiveness of existing vaccines

The development of novel vaccines and vaccine

delivery systems

These ongoing scientific advances in vaccine

development need to be accompanied by
scientific advances in epidemiological methods
which can continue to inform the development
and monitoring of new vaccines.