Types of Vaccines

Nonliving Vaccines
Vaccines may contain either living or killed organisms or purified antigens from these
organisms. Vaccines containing living organisms tend to trigger the best protective
responses. Killed organisms or purified antigens may be less immunogenic than living
ones because they are unable to grow and spread in the host. Thus, they are less likely to
stimulate the immune system in an optimal fashion. On the other hand, they are often less
expensive and may be safer. Living viruses from vaccines, for example, infect host cells
and grow briefly. The infected cells then process the viral antigens, triggering a response
dominated by cytotoxic T cells, a type 1 response. Killed organisms and purified antigens,
in contrast, commonly stimulate responses dominated by antibodies, a type 2 response.
This type of response may not generate optimal protection against some organisms. As a
result, vaccines that contain killed organisms or purified antigens usually require the use
of adjuvants to maximize their effectiveness. Adjuvants may, however, cause local
inflammation, and multiple doses or high doses of antigen increase the risks of producing
hypersensitivity reactions. Killed vaccines should resemble the living organisms as closely
as possible. Chemical inactivation should cause minimal change to their antigens.
Compounds used in this way include formaldehyde, ethylene oxide, ethyleneimine,
acetylethyleneimine, and beta-propiolactone.
Subunit Vaccines
Although vaccines containing whole killed organisms are economical to produce, they
contain many components that do not contribute to protective immunity. They may also
contain toxic components such as endotoxins. Thus, depending upon costs, it may be
advantageous to identify, isolate, and purify the critical protective antigens. These can
then be used in a vaccine by themselves. For example, purified tetanus toxin, inactivated
by treatment with formalin (tetanus toxoid), is used for active immunization against
tetanus. Likewise, the attachment pili of enteropathogenic Escherichia coli can be
purified and incorporated into vaccines. The antipilus antibodies protect animals by
preventing bacterial attachment to the intestinal wall.
Antigens Generated by Gene Cloning
The cost of physically purifying a specific antigen may be prohibitive. In such cases it
may be more appropriate to clone the genes coding for the protective antigens into a
vector such as a bacterium, yeast, baculovirus, or plant. The DNA encoding the desired
antigens may be inserted into its vector, which then expresses the protective antigen. The
recombinant vector is grown, and the antigens encoded by the inserted genes are
harvested, purified, and administered as a vaccine. An example of such a vaccine is one
directed against the cloned subunit of E coli enterotoxin. The cloned subunits are
antigenic and function as effective toxoids.
It is possible to clone viral antigen genes in plants. This has been successfully achieved for
viruses such as transmissible gastroenteritis virus and Newcastle disease virus. The plants
used include tobacco, potato, and corn. These plants contain very high concentrations of
antigen, and protection may be achieved by simply feeding the plants to animals.

DNA Plasmid Vaccines

Animals may also be immunized by injection of DNA encoding viral antigens. This DNA
is inserted into a bacterial plasmid, a piece of circular DNA that acts as a vector. When
the genetically engineered plasmid is injected, it is taken up by host cells. The DNA is
then transcribed, and mRNAs are translated to produce the vaccine protein.

This type of DNA plasmid vaccine is used to protect horses against  West Nile
virus infection. This approach has been applied experimentally to produce vaccines
against:
 avian influenza virus
 rabies virus in dogs and cats
 canine parvovirus
 bovine viral diarrhea virus
 feline immunodeficiency virus

 feline leukemia virus

  foot-and-mouth disease virus
 bovine herpesvirus-1 related disease

 Newcastle disease virus

Some DNA vaccines are able to induce immunity even in the presence of very high levels
of maternal antibody.

Modified Live Vaccines

Attenuated Vaccines
The use of live organisms in vaccines presents many advantages. For example, they are
usually more effective than inactivated vaccines in triggering cell-mediated immune
responses. Their use, however, also presents potential hazards. Thus, the virulence of a
live organism used for vaccination must be attenuated so that it is able to replicate but is
no longer pathogenic. The level of attenuation is critical to vaccine success.
Underattenuation will result in residual virulence and disease (reversion to virulence);
overattenuation will result in an ineffective vaccine. Rigorous reversion to virulence
studies must be performed to demonstrate stability. Attenuated vaccines should not be
used to vaccinate species for which they have not been tested or approved. Pathogens
attenuated for one species may be over- or under-attenuated in others. Thus, they may
either cause disease or fail to provide adequate protection.

For some diseases, related organisms normally adapted to another species may impart
limited immunity. Examples include vaccines against measles virus, which can protect
dogs against distemper, and against bovine viral diarrhea virus, which can protect pigs
against classical swine fever.

In rare circumstances, virulent organisms may be used for vaccination. The only current
example of this is vaccination against contagious ecthyma (Orf, sore mouth) of sheep.
Lambs are vaccinated by rubbing dried, infected scab material into scratches made on
the inner thigh, resulting in local infection and solid immunity. Because vaccinated
animals may spread the disease, however, they must be separated from unvaccinated
stock for a few weeks. Considerable care must also be exercised in the preparation,
storage, and handling of modified live vaccines to avoid temperature extremes that can
reduce the viability of the organisms. Likewise, vaccines such as Brucella strain RB51
and contagious ecthyma are zoonotic and present hazards to the administrator.

Attenuation of viruses by prolonged tissue culture can be considered a primitive form of

genetic engineering. Ideally, this resulted in the development of a strain of virus that was
unable to cause disease.

Another relatively simple method is to adapt the vaccine virus to grow at a temperature
approximately 10 degrees lower than normal body temperature. These cold-attenuated
vaccines can be administered intranasally, where they can grow in the cool upper
respiratory tract but not in the warmer lower respiratory tract or other organs.

Gene-deleted Vaccines
Molecular genetic techniques now make it possible to modify the genes of an organism so
that it becomes irreversibly attenuated. Deliberate deletion of the genes that code for
proteins associated with virulence is an increasingly attractive procedure. For example,
gene-deleted vaccines were first used against the Aujeszky disease herpesvirus. In this
case, the thymidine kinase gene was removed from the virus. Herpesvirus requires
thymidine kinase to return from latency. Viruses from which this gene has been removed
can infect neurons but cannot replicate and cause disease.

Similar genetic manipulation can also be used to restrict the ability of bacteria to grow in
vivo. For example, a modified live vaccine is available that contains streptomycin-
dependent Mannheimia haemolytica and Pasteurella multocida. These mutants depend on
the presence of streptomycin for growth. When used in a vaccine, the absence of
streptomycin will eventually result in the death of the bacteria, but not before they have
stimulated a protective immune response.
Virus-vectored Vaccines
Another way to produce a highly effective living vaccine is to insert the genes that encode
protective antigens into an avirulent “vector” organism. These vaccines are created by
deleting genes from the vector and replacing them with genes coding for antigens from
the pathogen. The recombinant vector is then administered as the vaccine, and the
inserted genes express the antigens when cells are infected by the vector virus. The vector
may be attenuated so that it will not be shed from the vaccinated animal, or it may be
host-restricted so that it will not replicate itself within the tissues of the vaccinate. Virus-
vectored vaccines are well-suited for use against organisms that are difficult or dangerous
to grow in the laboratory.

The most widely used vaccine viral vectors are large DNA viruses such as poxviruses,
vaccinia virus, adenoviruses, and some herpesvirus. These viruses have a large genome
that facilitates insertion of new genes. They also express relatively high levels of the
recombinant antigen. In at least some cases, vectored vaccines appear able to induce
immunity even when high levels of maternal antibody are present. Fowlpox virus and
herpesvirus recombinant vaccines are widely used in the poultry industry. For example,
one vector is fowlpox virus, into which Newcastle disease virus genes are incorporated. It
has the benefit of conferring immunity against fowlpox virus as well.

Vectored vaccines are commercially available for:

 avian influenza virus

 West Nile virus and influenza virus in horses

 feline leukemia virus

 vaccinating wildlife against rabies virus

These vaccines are safe, stable, can work in the absence of an adjuvant, and like the gene-
deleted vaccines, allow for differentiation from natural infections.