Developments in the Implementation of Vaccine Crops

Introduction
Vaccines are an effective but delicate method of disease prevention. Vaccine
production requires temperature controlled environments and highly regulated
processes and results in a product that possesses a limited effective lifespan.
This is compounded by the fact that very few sources of vaccine exist, with only
25 manufacturers being recognised by the WHO and the costs involved in
manufacture being prohibitive for small-scale production. Not only does this
present a challenge for distribution, but also for cost effectiveness and the
quantity and quality of the available supply (WHO, 2014).
As a result, many scientists are looking to find novel methods of synthesising the
antigens required for a vaccine. One method being examined is to create
transgenic crops that synthesise vaccines themselves, which could reduce the
cost of production and improve the ability to distribute and supply them (de
Vries, 2000).
Vaccine fundamentals
A vaccine works by introducing material that produces an immune response,
known as an antigen, into the body. These are given in sufficient quantity to be
detected by the immune system, without being so prevalent as to actually cause
illness. The cells that detect the antigens are lymphocytes classified as IgE B cell
precursors. Once activated, the cell differentiates into either an IgE antibodysecreting cell, which produce specific antibodies that bind to and help identify
the current antigen, or into a memory B cell, which are stored and can
differentiate into antibody-secreting cells should reinfection by the same antigen
occur again (Katz, 1980). In this way, the body is able to retain the ability to
respond to a library of known antigens, increasing the response time of the
immune system and increasing the likelihood of the infection being contained
easily.
The proposed idea for a vaccine crop would insert a gene into a plant genome
that would synthesise an antigen associated with a known pathogen. These
would be ingested like normal, non-vaccine strains of the plant and the antigen
would be absorbed through the digestive system. This method increases the
availability of the vaccine; the cost of a single dose of hepatitis vaccineexpressing banana has been estimated to be US$0.02; a vast reduction compared
to the equivalent vaccine injection at US$125(de Vries, 2000). In addition, it
allows production to be centred in the areas that require the vaccine (provided a
crop suitable for the climate is chosen), which should reduce the cost of
transportation and distribution.
So far, plants have successfully been modified to produce a variety of antigens,
targeting diverse diseases such as lymphoma (McCormick et al. 1999), Anthrax
infection (Watson et al. 2004) and Entamoeba histolytica infection (Chebolu &
Daniell, 2007). However, few of these cases are specific to food crops, with most
being experiments with model organisms, such as tobacco (Maliga, 2004).

Further study would be necessary to insert the antigen-producing genes into

edible crops, in order to make the vaccine easiest to administer.
Methods for gene integration
A matter of note is that many of these studies use chloroplasts as a gene vector to
carry the antigen-producing gene. Chloroplasts have several advantages in this
regard; as centres of metabolic activity, chloroplasts have high concentrations of
the amino acids required to construct proteins, allowing for efficient expression
of the antigen-producing gene. Furthermore, it prevents the gene from being
transmitted by pollen, preventing the gene from crossbreeding into regular
crops or wild strains and causing potential ecological damage in an uncontrolled
environment (Wang et al. 2009).
However, chloroplasts also carry a number of complications. Due to their
ubiquitous distribution throughout most plants, transgenic proteins expressed
by the chloroplasts would be produced in all tissues, rather than the specific
tissues required for their intended purpose. It has been suggested that this can
lead to excessive metabolic stress on the plant (due to the synthesis of
unnecessary protein) and ultimately reduce the viability of the plant (Magee et
al. 2004).
Other methods of inserting antigen-expressing genes include directly integrating
the gene into the plants genome through the use of Agrobacterium T-DNA
vectors or by directly bombarding the genome with microprojectiles (Mason and
Arntzen, 1995). As with the use of chloroplast vectors, these methods result in
permanent integration with the host, allowing for successive generations of
vaccine-producing crops to be produced, as well as granting the opportunity to
insert multiple genes to create more comprehensive vaccines. It also overcomes
the main drawback in the use of chloroplasts, as there is a potential to induce
tissue specific expression, provided the site of insertion is chosen carefully.
However, due to being integrated with the nuclear genome, these plants have the
potential to produce pollen bearing the antigen-expressing gene resulting in the
risk of contaminating wild populations as discussed earlier.
An alternative approach to vaccine production within crops is to avoid
permanent integration with the host genome, and instead use viral vectors to
introduce the antigen-producing gene into individual plants (Mason and Arntzen,
1995). Whilst this is far more labour-intensive approach (each individual plant
must be inoculated with the viral vector), there is a potential for greater yields of
antigen to be produced.
Choice of Crop
The plant chosen to carry the vaccine is of vital importance to its successful
distribution. In order to succeed, the crop must be capable of producing a enough
antigens to stimulate an immune response, without being toxic when ingested by
humans. Ideally, the crop should also be able to grow in the climatic conditions

present at their point of consumption, in order to keep distribution costs at a

minimum.
Tobacco has shown to be especially efficient at synthesising antigens, and is
capable of growing in a wide variety of climates (Watson et al. 2004), but is
obviously a non-food crop and carries all the risks associated with tobacco
consumption (increased risk of heart disease, exposure to carcinogens, etc) (NHS
smokefree, 2014).
A more likely candidate exists in the form of the potato plant (Mason and
Arntzen, 1995). Tubers from treated plants were shown to produce an immune
response when consumed by test subjects. It is of note that the effect was
produced even after the tubers were cooked for consumption. This would
increase the palatability of this method of treatment to potential patients,
improving popular approval of the treatment and increasing the likelihood of a
vaccination programme succeeding.
Conclusion
Crops cultivated to be used as edible vaccines show promise as a means of cost
effective vaccine distribution. Efforts to integrate individual antigen-producing
genes appear to have been successful, although it remains to be seen if more
complex combinations of antigens as used in current injection vaccines are
possible. Overall success hinges on being able to balance antigen yield per plant
with the ability to mass-produce transgenic plants without risking damage to
local ecosystems; a set of criteria that none of the current vector solutions
entirely fulfil. However, a carefully chosen crop specific to the local climate will
drastically improve the viability of any application of this technology, and will
allow for far cheaper access to vaccination in previously deprived areas.
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