VACCINE DEVELOPMENT, TESTING,

AND REGULATION

INTRODUCTION
The purpose of a vaccine is to provide protection against a pathogen (in our case a viral
pathogen). Early attempts at “vaccine” development were based on the observation that persons
surviving a disease, for example, smallpox, were protected against further occurrences of that
particular disease. Today we know that immune protection against a specific viral infection is
provided by “specialized” (also called “adaptive”) responses that inhibit virus replication or kill
infected cells. The goal of vaccination is to stimulate the body to develop a specialized,
protective, immune response in the absence of disease. Immune responses target one or more
viral proteins and there are a variety of ways that viral proteins can be delivered as vaccines.
These include administration of:
• Weakened (so-called attenuated) viruses that replicate in the host without causing disease.
• Inactivated (killed) virions.
• Purified protein products.
• Nucleic acids (genes) to direct synthesis of desired proteins.

CLASSICAL VERSUS ENGINEERED VACCINES

Traditional or classical methods to produce a viral vaccine required growing large quantities of
virus. Attenuated viruses were usually obtained by repeated passage in animals or cultured cells;
unfortunately the results were impossible to predict and some viruses were never adequately
attenuated even after prolonged passage. Today recombinant DNA technologies, along with our
understanding of the molecular details of virus replication and pathogenesis, provide an array of
opportunities to achieve vaccine goals. Recombinant DNA technologies can be used to make
targeted mutations in a virus or to clone and express specific viral proteins.

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Fig 1- General types of viral vaccines

As shown in fig 2, examples recombinant DNA strategies for vaccine development include:
• Virulent viruses can be weakened by deletion or modifications of specific genes.
• Specific genes from a virulent virus (often genes for capsid or envelope proteins) can be
transferred to a safer virus. The recombinant virus might be administered as a replicating virus or
might be grown and inactivated.
• Specific genes from a virulent virus might be moved into bacteria, yeast, or cultured cells to
produce large quantities of a particular viral protein.
• DNA molecules encoding specific proteins, or parts of proteins might be generated and
administered directly to the patient.
In addition, the use of adjuvants (substances that modify the effects of other agents) provides
even more flexibility, as there are a wide variety of adjuvants that can enhance or modify host
immune responses.

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Fig 2- A variety of methods can be used to produce genetically engineered vaccines.

Vaccine development is a long, complex process, often lasting 10-15 years and involving a
combination of public and private involvement.

The current system for developing, testing, and regulating vaccines developed during the
20th century as the groups involved standardized their procedures and regulations.

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 STAGES OF VACCINE DEVELOPMENT AND TESTING

Delivery of a vaccine in a programme such as Expanded Program on Immunization is the end

In the United States, vaccine development and testing follow a standard set of steps. The first
stages are exploratory in nature. Regulation and oversight increase as the candidate vaccine
makes its way through the process.

Development of vaccines can be simplified into two broad stages:

1. Pre-clinical development is research carried out in lab assays and on animals. It

 Identification (discovery) of relevant antigens (e.g. screening)

 Creation of the vaccine concept

 Evaluation of vaccine efficacy in test tubes and animals

 Manufacture of the vaccine to Good Manufacturing Practice standards

2. Clinical development is when the vaccine is first tested in humans. It covers four stages
over several years, from initial clinical trials in humans (phase I) right through to introduction
and beyond (phase IV). Clinical development is built on rigorous ethical principles of informed
consent from volunteers, with an emphasis on vaccine safety as well as efficacy.

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First Steps: Laboratory and Animal Studies

Exploratory Stage

This stage involves basic laboratory research and often lasts 2-4 years. Federally funded
academic and governmental scientists identify natural or synthetic antigens that might help
prevent or treat a disease. These antigens could include virus-like particles, weakened viruses or
bacteria, weakened bacterial toxins, or other substances derived from pathogens.

Pre-Clinical Stage

Pre-clinical studies use tissue-culture or cell-culture systems and animal testing to assess the
safety of the candidate vaccine and its immunogenicity, or ability to provoke an immune
response. Animal subjects may include mice and monkeys. These studies give researchers an
idea of the cellular responses they might expect in humans. They may also suggest a safe starting
dose for the next phase of research as well as a safe method of administering the vaccine.

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 the animals, meaning that they vaccinate the
animals and then try to infect them with the target pathogen.

Many candidate vaccines never progress beyond this stage because they fail to produce the
desired immune response. The pre-clinical stages often lasts 1-2 years and usually involves
researchers in private industry.

IND Application

A sponsor, usually a private company, submits an application for an Investigational New Drug
(IND) to the U.S. Food and Drug Administration. The sponsor describes the manufacturing and
testing processes, summarizes the laboratory reports, and describes the proposed study. An
institutional review board, representing an institution where the clinical trial will be conducted,
must approve the clinical protocol. The FDA has 30 days to approve the application.

Once the IND application has been approved, the vaccine is subject to three phases of testing.

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Next 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 of the test subjects until they reach their target.
Phase I trials may be non-blinded (also known as open-label in that the researchers and perhaps
subjects know whether a vaccine or placebo is used).

The goals of Phase 1 testing are to assess the safety of the candidate vaccine and to determine the
type and extent of immune response that the vaccine provokes. In a small minority of Phase 1
vaccine trials, researchers may use the challenge model, attempting to infect participants with the
pathogen after the experimental group has been vaccinated. The participants in these studies are
carefully monitored and conditions are carefully controlled. In some cases, an attenuated, or
modified, version of the pathogen is used for the challenge.

A promising Phase 1 trial will progress to the next stage.

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 and involve the
experimental vaccine being tested against a placebo (the placebo may be a saline solution, a
vaccine for another disease, or some other substance).

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One Phase III goal 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. For example, suppose
that an adverse event related to a candidate vaccine might occur in 1 of every 10,000 people. To
detect a significant difference for a low-frequency event, the trial would have to include 60,000
subjects, half of them in the control, or no vaccine, group (Plotkin SA et al. Vaccines, 5th ed.
Philadelphia: Saunders, 2008).

Vaccine efficacy is tested as well. These factors might include 1) Does the candidate vaccine
prevent disease? 2) Does it prevent infection with the pathogen? 3) Does it lead to production of
antibodies or other types of immune responses related to the pathogen?

Next Steps: Approval and Licensure

After a successful Phase III trial, the vaccine developer will submit a Biologics License
Application to the FDA. Then the FDA will inspect the factory where the vaccine will be made
and approve the labeling of the vaccine.

After licensure, the FDA will continue to monitor the production of the vaccine, including
inspecting facilities and reviewing the manufacturer’s tests of lots of vaccines for potency, safety
and purity. The FDA 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. They include Phase IV
trials, the Vaccine Adverse Event Reporting System, and the Vaccine Safety Datalink.

Phase IV Trials

Phase IV trial are optional studies that drug companies may conduct after a vaccine is released.
The manufacturer may continue to test the vaccine for safety, efficacy, and other potential uses.

VAERS

The CDC and FDA established The Vaccine Adverse Event Reporting System in 1990. The goal
of VAERS, according to the CDC, is “to detect possible signals of adverse events associated with

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vaccines.” (A signal in this case is evidence of a possible adverse event that emerges in the data
collected.) About 30,000 events are reported each year to VAERS. Between 10% and 15% of
these reports describe serious medical events that result in hospitalization, life-threatening
illness, disability, or death.

VAERS is a voluntary reporting system. Anyone, such as a parent, a health care provider, or
friend of the patient, who suspects an association between a vaccination and an adverse event
may report that event and information about it to VAERS. The CDC then investigates the event
and tries to find out whether the adverse event was in fact caused by the vaccination.

The CDC states that they monitor VAERS data to

 Detect new, unusual, or rare vaccine adverse events

 Monitor increases in known adverse events

 Identify potential patient risk factors for particular types of adverse events

 Identify vaccine lots with increased numbers or types of reported adverse events

 Assess the safety of newly licensed vaccines

Not all adverse events reported to VAERS are in fact caused by a vaccination. The two
occurrences may be related in time only. And, it is probable that not all adverse events resulting
from vaccination are reported to VAERS. The CDC states that many adverse events such as
swelling at the injection site are underreported. Serious adverse events, according to the CDC,
“are probably more likely to be reported than minor ones, especially when they occur soon after
vaccination, even if they may be coincidental and related to other causes.”

VAERS has successfully identified several rare adverse events related to vaccination. Among
them are

 An intestinal problem after the first vaccine for rotavirus was introduced in 1999

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  Neurologic and gastrointestinal diseases related to yellow fever vaccine

Additionally, according to Plotkin et al., VAERS identified a need for further investigation of
MMR association with a blood clotting disorder, encephalopathy after MMR, and syncope after
immunization (Plotkin SA et al. Vaccines, 5th ed. Philadelphia: Saunders, 2008).

Vaccine Safety Datalink

The CDC established this system in 1990. The VSD is a collection of linked databases
containing information from large medical groups. The linked databases allow officials to gather
data about vaccination among the populations served by the medical groups. Researchers can
access the data by proposing studies to the CDC and having them approved.

The VSD has some drawbacks. For example, few completely unvaccinated children are listed in
the database. The medical groups providing information to VSD may have patient populations
that are not representative of large populations in general. Additionally, the data come not from
randomized, controlled, blinded trials but from actual medical practice. Therefore, it may be
difficult to control and evaluate the data.

Rapid Cycle Analysis is a program of the VSD, launched in 2005. It monitors real-time data to
compare rates of adverse events in recently vaccinated people with rates among unvaccinated
people. The system is used mainly to monitor new vaccines. Among the new vaccines being
monitored in Rapid Cycle Analysis are the conjugated meningococcal vaccine, rotavirus
vaccine, MMRV vaccine, Tdap vaccine, and the HPV vaccine. Possible associations between
adverse events and vaccination are then studied further.

REVERSE VACCINOLOGY AND APPROACHES TO VACCINE DEVELOPMENT

There are two different approaches available for vaccine development. In the conventional
approach the microorganism responsible for causing a disease is first cultivated (cultured) in a
laboratory following which the microorganism is analyzed and various antigens of the organism
are selected as targets for development of the vaccine. These antigens are then used for
immunogenicity testing in animal models, and among the various antigens one will be selected
for vaccine development. The identification of protective antigens that could be potential vaccine

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candidates potentially involves the separating of each component one at a time. This approach is
time consuming and allows the identification of only those antigens that can be purified in
quantities suitable for vaccine testing.

The most abundant proteins of a microorganisms are sometimes not suitable vaccine candidates
(Sodium Potassium pump). While the tools required to identify the less abundant components
may be inadequate or not available at all because of which the conventional approach can take
years or decades. Moreover, many pathogens cannot be grown in laboratory conditions because
of which developing a vaccine by the conventional approach becomes impossible.

Hence, the reverse vaccinology approach was introduced in the search for more efficient
vaccines. This approach takes advantage of the genome sequence of the pathogen. By computer
analysis, the genome sequence is studied and predictions are made as to the most likely vaccine
candidates. This approach thus allows the identification of novel antigens that usually cannot be
detected by the conventional system because of their low abundance. Once the vaccine
candidates have been identified they can be used for the preparation of DNA vaccines.

Government Oversight

In the United States

At the end of the 19th century, several vaccines for humans had been developed. They were
smallpox, rabies, plague, cholera, and typhoid vaccines. However, no regulation of vaccine
production existed.

On July 1, 1902, the U.S. Congress passed "An act to regulate the sale of viruses, serums, toxins,
and analogous products," later referred to as the Biologics Control Act (even though "biologics"
appears nowhere in the law). This was the first modern federal legislation to control the quality
of drugs. This act emerged in part as a response to 1901 contamination events in St. Louis and
Camden involving smallpox vaccine and diphtheria antitoxin.

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The Act created the Hygienic Laboratory of the U.S. Public Health Service to oversee
manufacture of biological drugs. The Hygienic Laboratory eventually became the National
Institutes of Health. The Act established the government’s right to control the establishments
where vaccines were made.

The United States Public Service Act of 1944 mandated that the federal government issue
licenses for biological products, including vaccines. After a poliovirus vaccine accident in 1954
(known as the Cutter incident), the Division of Biologics Standards was formed to oversee
vaccine safety and regulation. Later, the DBS was renamed the Bureau of Biologics, and it
became part of the Food and Drug Administration. It is now known as the Center for Biologics
Evaluation and Research.

Elsewhere

In the European Union, the European Medicines Agency supervises regulation of vaccines and
other drugs. A committee of the World Health Organization makes recommendations for
biological products used internationally. Many countries have adopted the WHO standards.

CONCLUSION

Vaccines are developed, tested, and regulated in a very similar manner to other drugs. In general,
vaccines are even more thoroughly tested than non-vaccine drugs because the number of human
subjects in vaccine clinical trials is usually greater. In addition, post-licensure monitoring of
vaccines is closely examined by the Centers for Disease Control and the FDA.

REFERENCES

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https://www.historyofvaccines.org/index.php/content/articles/vaccine-development-testing-and-
regulation

https://www.vaccineseurope.eu/about-vaccines/key-facts-on-vaccines/how-are-vaccines-developed/

http://www.euvaccine.eu/vaccines-diseases/vaccines/stages-development

https://www.sciencedirect.com/science/article/pii/B9780128031094000076?via%3Dihub

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