Dire dawa University
College of Medicine and health sciences
Department of Medical Laboratory Sciences
Chapter -10
Vaccinology
Outline
Definition
Stages of vaccine development
Classification of vaccines
Vaccine Adjutants
Definition of Vaccinology
Vaccinology is the science of developing vaccines to
prevent or treat diseases.
A vaccine is a biological preparation that improves
immunity to a particular disease.
A vaccine typically contains an agent that resembles
a disease-causing microorganism and is often made
from weakened or killed forms of the microbe, its
toxins or one of its surface proteins.
The agent stimulates the body's immune system to
recognize the agent as foreign, destroy it, and keep a
record of it, so that the immune system can more easily
recognize and destroy any of these microorganisms that
it later encounters.
Vaccinology in the last two decades
The application of molecular genetics and its
increased insights into Immunology, Microbiology
and Genomics applied to vaccinology.
Molecular genetics sets the scene for a bright future
for vaccinology including:
The development of new vaccine delivery systems
(e.g. DNA vaccines, viral vectors)
New adjuvants,
The development of more effective vaccines,
New vaccines against existing and emerging disease
Therapeutic vaccines may also soon be available for
cancer, allergies, autoimmune diseases and
addictions.
Why vaccine and vaccination?
Even with the significant advancements in modern
medicine
vaccination continues to be the most cost-effective
method to reduce economic loss and suffering
from infectious diseases
Vaccines can be:
Prophylactic - prevent the effects of a future
infection by any natural pathogen.
Therapeutic –to treat existing disease.
Active and Passive Immunization
Active Immunization
Many diseases stimulate an immune response in host, those who
survive the disease are protected from second infection – natural
acquired active immunization
- the risk is many die before becoming immune
Vaccinations uses
Artificially acquired active immunity - is stimulated by initial
exposure to specific foreign macromolecules through the use of
vaccines, to artificially establish state of immunity.
Individuals, who have not had the disease, can be protected even
when exposed at later date
Active and Passive Immunization
Limitations of Active immunity
developing an immune response does not = achieving
state of protective immunity
vaccine can induce primary response but fail to induce
memory cells = host unprotected
Active and Passive Immunization
Ideal Vaccination Response
Passive Immunization
Artificially acquired passive immunity; the direct transfer of
antibodies to a non-immune person to provide temporary
protection. Like
molecules (antibodies)-Tetanus Anti-toxoid or
cell (lymphocytes) produced in another individual
Naturally acquired passive immunity (maternal Abs) - refers to
antibodies transferred from mother to fetus across the placenta
and to the newborn in colostrum and breast milk during the first
few months of life.
Passive immunization does not activate the immune system, it
generates no memory response and the protection provided is
transient.
Stages of Vaccine Development and Testing
Vaccine development and testing follow a standard set of
steps.
On average, it takes 10-15 years to bring a vaccine to
market.
Vaccine development is highly pyramidal in shape; for
each success there are many failures.
Most failures occur in pre-clinical stage and in early phase
1 clinical trial.
Stages…
Implementation
Licensure
Phase 3
Phase 2
Phase 1
Pre-Clinical
Pyramidal nature of vaccine development
First Steps: Laboratory and Animal Studies
Exploratory Stage
This stage involves basic laboratory research and often
lasts 2-4 years.
Researchers identify natural or synthetic antigens that
might help prevent or treat a disease.
First Steps: Laboratory and Animal Studies
Pre-Clinical Stage
Pre-clinical studies use cell culture systems and animal
testing to assess the safety and immunogenicity
These studies give researchers an idea of the cellular
responses they might expect in humans.
Researchers may adapt the candidate vaccine during
the pre-clinical state to try to make it more effective.
They may also do challenge studies with animals
Challenge studies are never conducted in humans.
Second Steps: Clinical Studies with Human Subjects
Phase I Vaccine Trials
This first attempt to assess the candidate vaccine in
humans involves a small group of adults, usually
between 20-80 subjects.
If the vaccine is intended for children, researchers
will first test adults, and then gradually step down the
age
The goals of Phase 1 testing are to:
assess the safety of the candidate vaccine
determine the type and extent of immune response
that the vaccine provokes.
A promising Phase 1 trial will progress to the next
phase.
Phase II Vaccine Trials
A larger group of several hundred individuals
participates in Phase II testing.
Some of the individuals may belong to groups at risk
of acquiring the disease.
These trials are randomized and well controlled, and
include a placebo group.
The goals of Phase II testing are to:
Study the candidate vaccine’s safety,
Immunogenicity,
Proposed doses,
Schedule of immunizations, and
Method of delivery.
Phase III Vaccine Trials
Successful Phase II candidate vaccines move on to
larger trials,
Involving thousands to tens of thousands of people.
These Phase III tests are randomized and double
blind.
The goal of Phase III testing is to assess vaccine
safety in a large group of people.
Certain rare side effects might not surface in the
smaller groups of subjects tested in earlier phases.
Third Steps: Approval and Licensure
After a successful Phase III trial, the vaccine
developer will submit a Biologics License
Application to the regulatory authority.
Then the regulatory authority will inspect the factory
approve the labeling of the vaccine.
After licensure, the regulatory body will continue to
monitor the production of the vaccine,
The regulatory body has the right to conduct its own
testing of manufacturers’ vaccines.
Post-Licensure Monitoring of Vaccines
A variety of systems monitor vaccines after they have
been approved. These include:
Phase IV trials,
The Vaccine Adverse Event Reporting System
(VAERS),
etc.
Phase IV Trials
Phase IV trials are optional studies that the companies
may conduct after a vaccine is released.
The manufacturer may continue to test the vaccine for
safety, efficacy, and other potential uses.
Some parameters of an ideal vaccine
The primary concern in vaccination is
Efficacy and safety.
Thus, vaccination itself must not cause any adverse
reactions and should result in greater than 90% efficacy
after a single administration regardless of species or type
of vaccine.
Long-lived immunity (hopefully life-long)
Effective when given orally (no need for injections)
Induces a wide range of appropriate responses (mucosal,
humoral, cellular)
Low cost
Compatible with local management practices
Compatible with co-administration of other vaccines
Stable (genetically/thermally)
Individual Vs Community immunity
Individual immunity-Vaccination’s immediate benefit
It provides long-term, sometimes lifelong protection
against a disease.
Community immunity- refers to the protection offered to
everyone in a community by high vaccination rates=Herd
immunity
With enough people immunized against a given disease,
it’s difficult for the disease to gain a foothold in the
community.
Antigen selection for vaccine development
Antigen selection is a critical step in the development effective
vaccine
Key considerations for selecting antigens for vaccine prep:
1. Conservation 5. Stability
2.Immunogenicity
6. Accessibility
3. Functionality
7. Cross-reactivity
4. Safety
6/10/2024 Seminar on Antigen for Dengue vaccine 24
Classification of vaccines
Genetic Vaccine
Whole organism vaccine
Subunit vaccine
Live attenuated DNA, RNA
viral
bacterial
Inactivated Vaccines
viruses protein-based polysaccharide-based
bacteria toxoid pure
Recombinant prot conjugate 25
Whole organism Vaccine
Attenuated virus or bacteria are used as a vaccine.
Microbes lose their pathogenicity but retain their capacity for
transient growth within an inoculated host.
The immune response to a live attenuated vaccine is virtually
identical to that produced by a natural infection.
The immune system does not differentiate between an infection
with a weakened vaccine virus and an infection with a wild virus.
Live attenuated vaccines produce immunity in most recipients with
one dose, except those administered orally.
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Attenuation
Growing a pathogen under abnormal culture conditions
E.g. BCG was render avirulent by culturing M.bovis on
bile salt saturated medium for 13 years
Prolonged virus culture with the cells to which it is not
adapted.
Passaging human pathogen through different hosts for
prolonged time. This method was used for virus.
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Advantages: attenuated vaccines
High immunogenicity- because live vaccines infect hosts and
replicate transiently- low dose is required
May not need adjuvants and can be applied by natural route of
infection.
Prolonged immune response
Single immunization may suffice for protection
Capable of inducing cellular & humoral immune response
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Disadvantages of live vaccine
Possibility to revert into a virulent form. Pathogens may
mutate and regain their virulence. E.g. Sabin polio
vaccine =1 per 2.4 million
Reversion implies the possibility of pathogenic forms
being passed into the natural environment.
May be associated with complications; E.g. measles
vaccine develop post-vaccine encephalitis or other
complications.
Low stability (short shelf -life)
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Inactivated whole organism vaccine
Inactivated vaccines are not alive and cannot replicate.
These vaccines cannot cause disease from infection, even in an
immunodeficient person.
Generally not as effective as live vaccines
Generally require 3-5 doses
Immune response mostly humoral
Antibody titer may diminish with time
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Methods of inactivation
Killed organisms for vaccine are expected to remain as antigenic as the
live organism
Inactivation of the pathogen by heat or by chemical means were
common approches of vaccine productions.
Heat inactivation is generally unsatisfactory because it causes extensive
denaturation of proteins
Some of the chemicals used in synthesis of inactivated vaccines are:
Formaldehyde: cross-link proteins and nucleic acids, and confers
structural rigidity. The Salk polio vaccine is produced by formaldehyde
inactivation.
Alkylating agents: such as ethyleneoxide, ethyleneamine, β-
propiolaceton cross-links nucleic acid chains and leave surface protein
unchanged.
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Merits of inactivated vaccines
Stable on storage
Do not replicate; and not likely to spread to other
organisms
Safe in immune-deficient patients
Low production costs
But it has some limitations
Proper booster doses are required and
Higher doses are required to induce long-term immunity.
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Inactivated exotoxin (toxoid)
bacterial pathogens, including those causing diphtheria and
tetanus, produce exotoxins…. Virulent factors.
Diphtheria and tetanus vaccines can be made by:
purifying the bacterial exotoxin,
inactivating the toxin with formaldehyde to form a toxoid.
Vaccination with the toxoid induces anti-toxoid antibodies,
which bind to the toxin and neutralizing its effects.
toxoid production must be closely controlled to achieve
detoxification without excessive modification of the epitope
structure.
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Recombinant protein vaccines
• A gene encoding any immunogenic protein can be cloned
and expressed in bacterial, yeast, or mammalian cells
using recombinant DNA technology.
• the expressed antigens are purified and used for vaccine
development.
• hepatitis B vaccine - vaccine was developed by cloning
the gene for the major surface antigen (HBsAg) and
expressing it in yeast cells.
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Pro and cons of rProt vaccine
Pros:
• Avoid the risks associated with attenuated vacc.
• Fairly stable
• Induce memory response
Cons
• Elicits humoral response predominantly
• Possibility of contamination
• Change on native conformation, so that antibodies
produced against the subunit may not recognize the same
protein on the pathogen surface
• Practical complexity
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Conjugate Vaccines
A conjugate vaccine is created by covalently attaching a poor
(polysaccharide organism) antigen to a carrier protein (preferably from the
same microorganism).
This technique for the creation of an effective immunogen is most often
applied to bacterial polysaccharides.
Polysaccharide coatings disguise a bacterium’s antigens so that the
immature immune systems of infants and younger children can’t recognize
or respond to them.
Conjugate vaccines are a special type of subunit vaccine to get around this
problem.
Haemophilus influenzae type B (Hib) is a conjugate vaccine.
DNA Vaccines
How do gene vaccines work?
Following in vivo transfection of host cells with plasmid DNA:
DNA is taken up by cells and subsequently enters the nucleus, where
mRNA of the respective antigen is transcribed and shuttled back into the
cytoplasm with subsequent translation of the gene product.
A finite amount of these newly synthesized proteins is degraded within
the cytoplasm by the proteasome and the resulting peptides will be
presented to CD8+ T cells in association to MHC I molecules.
On the other hand, exogenous antigens secreted out from transfected
cells or released out upon death of these cells, due to the transfection
event, are taken up by APCs like DCs.
These cells process the internalized antigen and present the resulting
peptides to CD4+ T cells in association with MHC II molecules.
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Advantages of DNA vaccines
free of the problem associated with producing
recombinant protein vaccines such as
improper folding of the gene product,
or complex biochemical procedures for protein
expression and purification.
DNA vaccines deliver genetic information …..
Expression under physiological conditions.
no problem of batch to batch variation in quality of the
resulting protein.
Induce both cellular and humoral response
stable
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Vaccine Adjuvants
Adjuvants
Enhance the immunogenicity of antigens
Modify the nature of the response
Reduce the antigen amount needed
Reduction frequency of booster immunization
Improve immune response in elderly and
immunocompromised vaccinees.
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