Vaccines

Vaccines….
Vaccine Design
 should be designed to induce the same immune responses that respond
to an actual infection and stimulate immunity to the microbial
antigens.
 An Ideal Vaccine:
 Should promote resistance to the disease

 Should induce long-lived memory.

 Should be safe, without side effects or allergic responses.

 Should be stable during storage and shipping.

 Should be inexpensive to control disease in developing counties.

Vaccines….
Antigens and Epitopes
 Antigens are macromolecules that elicit an immune response in the body.

 Antigens can be proteins, polysaccharides or conjugates of lipids with

proteins (lipoproteins) and polysaccharides (glycolipids).
 Antigens are generally very large and complex and the lymphocytes may
not recognize all the sites of a particular antigen.
 Rather both B and T lymphocytes recognize discreet sites on an antigen
known as epitopes or antigenic determinants.
 The aim in vaccine production is to ensure that epitopes exist on the
vaccine which will elicit humoral or cell-mediated response.
Vaccines….
Types of Vaccines and Their Characteristics
 There are now several basic types of vaccine.

Live attenuated vaccines

 In live attenuated vaccines, the organism has been cultured so as to
reduce its pathogenicity, but still retains some of the antigens of the
virulent form.
 They consist of the living pathogens whose virulence has been
reduced or attenuated.
Vaccines….
Attenuation
 Attenuation is any process that substantially lessens or negates the
virulence of viruses or bacteria.
 Usually achieved by modifying the growth conditions or manipulating
microbial genes in a way that eliminates virulence factors.
 Methods include

1. long-term cultivation,

2. selection of mutant strains that grow at colder temperatures (cold mutants),

3. passage of the microbe through unnatural hosts or tissue culture, and

4. removal of virulence genes.

Vaccines….
Example
 The vaccine for tuberculosis (BCG) was obtained after 13 years of
subculturing the agent of bovine tuberculosis.
 Vaccines for measles, mumps, polio (Sabin), and rubella contain live,
nonvirulent viruses.

Advantages
(1) Viable microorganisms can multiply and produce infection (but not
disease) like the natural organism;

(2) they confer long-lasting protection; and

(3) they usually require fewer doses and boosters than other types of vaccines.
Vaccines….
Disadvantages
1. they require special storage facilities,

2. can be transmitted to other people, and

3. can mutate back to a virulent strain

 Attenuated vaccines are not recommended for people whose immune
systems are compromised.
 If available, inactivated vaccines are substituted.
Vaccines….
Inactivated whole-agent vaccines/Killed vaccines
Consist of suspensions of fully virulent organisms (bacteria or viruses)
killed as mildly as possible in order not to destroy the antigenic
determinants on the organism.
Killing can be achieved by

heat (usually about 60°C for 1 hour)

chemicals (phenol, alcohol, formalin, β-propiolactone) or

ultraviolet irradiation.
Vaccines….
Examples
TAB vaccine against typhoid fever and which consists of heat-killed
phenol-preserved suspension of Salmonella typhii and S. Paratyphii A &
B, whooping cough, cholera, and the Salk IPV.

Disadvantages
Killed vaccines do not provide as prolonged antigenic stimuli as living
vaccines and
two, three or more subcutaneous injections are required to give
adequate protection.
Vaccines….
Vaccines….
 Bacterial toxoid
 In some diseases such as diphtheria and tetanus, the bacterial metabolite (a
protein toxin) is the cause of the disease and not the bacteria themselves.
 Toxoids are inactivated bacterial exotoxins.

 Toxoids induce antibody production when injected into the body, although
they are themselves harmless.
 Exposing the toxin with formaldehyde, denatures the protein.

 However, some epitopes on the protein molecule are retained and they
elicit antibody production.
 They require a series of injections for full immunity, followed by
boosters every 10 years.
 Subunit vaccine
 Subunit vaccines contain antigens or epitopes that induce protection rather
than the whole organism.
 The materials usually come from the surface of the organism and hence
they are also known as surface molecule vaccines.
 Several vaccines employ purified surface molecules.

 One of them, the influenza vaccine contains purified hemagglutinins from

the viruses.
 Another example is vaccine against hepatitis B virus.

 The gene encoding a protein expressed on the surface of the virus, the B
surface antigen or HBsAg, can now be expressed in E. coli cells and
provides the material for an effective vaccine.
 Subunit vaccine
 The potential advantages of using subunits as vaccines are

the increased safety and

less antigenic competition (since only a few components are included
in the vaccine).
 Disadvantages of subunit vaccines are

they generally require strong adjuvants and these adjuvants often
induce tissue reactions.
• (Adjuvants are compounds administered with vaccines so as to increase the
immunogenicity of the vaccines.)


the duration of immunity is generally shorter than with live vaccines.
Sometimes peptide epitopes may be used.
 Conjugate vaccine
 These are similar to subunit vaccines in the sense that only a part of the
organism is used in making the vaccine.
 Some bacteria which are encapsulated cause important childhood diseases
such as septicemia, pneumonia and meningitis.
 The bacteria are Hemophilus influenzae type B (HiB), Neissseria
meningitides and Streptococuus pneumoniae.
 The capsules of these bacteria are made of carbohydrates which the
immune system of adults recognize as foreign, but which that of infants
do not and hence cannot make antibodies against them.
 To solve the problem protein from diphtheria or tetanus toxoids is linked
or conjugated to the carbohydrate to make a vaccine.
 Conjugate vaccine
 This enables a baby’s immune system to respond to the combined
vaccine and produce antibodies.
 The licensed conjugate vaccines against Haemophilus influenzae type
b (Hib), previously the major cause of bacterial meningitis in babies
and young children, have virtually eliminated the disease in the
United States.
 Peptide vaccine
 It is possible to identify the epitopes within a protein that can
induce neutralizing antibodies or epitopes that are important in T-
cell responses to vaccines.
 Chemical synthesis of these epitopes is relatively straightforward.
 With an appropriate adjuvant or conjugation to carrier proteins, they
can induce antibody or T-cell mediated responses to the synthesized
epitope.
 These immune responses are in some cases sufficient to give
protection against the organism the protein epitope was derived from.
 Advantages of this approach include

the product is stable and chemically defined without the presence
of an infectious agent and

can be designed to stimulate specific T- and B-cell responses.

are relatively simple and

require limited work-up and purification.
 peptide vaccine
 The major disadvantage for peptide vaccines is based on the fact that
often only one peptide is used.
 Many pathogens are characterized by the fact that there is
extreme variation in the antigenic proteins of the agent.
 A single epitope or even multiple epitopes are often insufficient for
protection against extreme variation of an agent such as HIV or
hepatitis C virus.
 Mutants can also arise, particularly for RNA viruses, and hence
allow growth of the pathogen because the immunity generated by a
single peptide cannot recognize the mutant.
 Genetic immunization
 A recent development in vaccinology is immunization with
polynucleotides.
 Also known as genetic immunization or DNA immunization.

 Plasmids of "naked" DNA injected into muscle results in the production

of the protein antigen encoded in the DNA.
 The injection can be made by conventional needle or, more efficiently, by
the "gene gun" method.
 DNA vaccines for humans are still in the early phases of testing.

 Two DNA vaccines for animals have been approved; one that
protects horses from West Nile virus, and another that protects
salmon from a serious viral disease.
 Advantages of polynucleotide immunization are that

it is extremely safe,

induces a broad range of immune responses (cell-mediated and
humoral responses),

long-lived immunity, and,

the animal body itself produces the vaccine which makes the
vaccine relatively inexpensive to produce.
 Recombinant vaccine vectors
 Genetically engineered viral or bacterial vectors that express antigens of interest.

 These vectors are often employed when trying to stimulate CD8+CTL responses.
 BCG
 The vaccine used to immunize against tuberculosis was derived
after 13 years’ passage in bile-containing medium by Calmette and
Guerin (hence the name BCG - bacille Calmette-Guerin).
 Recombinant BCG Vaccines. BCG is an avirulent bovine tubercle
bacillus that is the most widely used vaccine in the world.
 Since 1948 over 5 billion vaccinations have been carried out.

 It is possible to introduce foreign DNA into BCG to express

antigens from other organisms, for example, the envelope
glycoprotein of human immunodeficiency virus (HIV).
 BCG
 Advantages include

A single immunization with BCG gives long-lasting cell-
mediated immunity to tuberculosis,

it can be given repeatedly,

is very safe with less than one complication per million
vaccinations, and

is a highly potent adjuvant.
 BCG and oral polio vaccine are the only two vaccines WHO
recommend to be given at birth.