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.
THANK YOU