Recombinant Vaccines

Revolutionizing Immunization
 Recombinant Vaccines
Recombinant DNA technology in recent years has
become a boon to produce new generation
vaccines. By this approach, some of the limitations
of traditional vaccine production could be
overcome. In addition, several new strategies,
involving gene manipulation are being tried to
create innovative recombinant vaccines.
Types of Recombinant Vaccines
Subunit recombinant vaccines:
These are the components of the pathogenic organisms. Subunit
vaccines include proteins, peptides and DNA.
Attenuated recombinant vaccines:
These are the genetically modified pathogenic organisms (bacteria or
viruses) that are made non-pathogenic and used as vaccines.
Vector recombinant vaccines:
These are the genetically modified viral vectors that can be used as
vaccines against certain pathogens. Some of the developments made in
the production of recombinant vaccines against certain diseases are
briefly described.
Subunit Vaccines:
As already stated, subunit recombinant vaccines
are the components (proteins, peptides, DNAs) of
the pathogenic organisms. The advantages of
these vaccines include their purity in preparation,
stability and safe use. The disadvantages are —
high cost factor and possible alteration in native
conformation. Scientists carefully evaluate the pros
and cons of subunit vaccines for each disease, and
proceed on the considered merits.
 DNA Vaccines
• Genetic immunization using DNA vaccines is a novel approach that came into being in 1990.
The immune response of the body is stimulated by a DNA molecule. A DNA vaccine consists
of a gene encoding an antigenic protein, inserted onto a plasmid, and then incorporated into
the cells in a target animal.
• The plasmid carrying DNA vaccine normally contains a promoter site, cloning site for the
DNA vaccine gene, origin of replication, a selectable marker sequence (e.g. a gene for
ampicillin resistance) and a terminator sequence (a poly—A tail).
• DNA vaccine—plasmids can be administered to the animals by one of the following delivery
methods.
• i. Nasal spray
• ii. Intramuscular injection
• iii. Intravenous injection
• iv. Intradermal injection
• v. Gene gun or biolistic delivery (involves pressure delivery of DNA-coated
gold beads).
Humoral immunity:
✓As the antigen molecules bind to B- lymphocytes, they trigger the
production of antibodies which can destroy the pathogens. Some of
the B-lymphocytes become memory cells that can protect the host
against future infections.
Cellular immunity:
✓The protein fragments of the antigen bound to MHC molecules can
activate the cytotoxic T-lymphocytes. They are capable of destroying
the infected pathogenic cells. Some of the activated T-lymphocytes
become memory cells which can kill the future infecting pathogens.
Complementary DNA vaccines:
✓For genetic immunization, complementary DNA (cDNA) vaccines can
also be used. Some workers have successfully used cDNA as
vaccines e.g. immunization of mice against influenza.
 Advantages of DNA vaccines
There are several advantages of using DNA vaccines in immunization:
✓1. The tedious and costly procedures of purifying antigens or
creating recombinant vaccines are not necessary.
✓2. DNA vaccines are very specific in producing the target proteins
(antigens or antibodies). Thus, they trigger immune response only
against the specific pathogen.
✓3. In general, DNA vaccines elicit much higher immune response
compared to other kinds of vaccines.
✓4. DNA vaccines are more stable for temperature variations (low or
high) than the conventional vaccines. Thus, the storage and transport
problems associated with vaccines are minimal.
✓5. The delivery methods to the host are simpler for DNA vaccines.
 Disadvantages of DNA vaccines:
✓1. The fate of the DNA vaccine in the host cells is
not yet clear. There is a possibility of this DNA
getting integrated into the host genome and this
may interrupt the normal functions.
✓2. There also exists a danger of cancer due to
DNA vaccines.
✓3. The post-translational modification of the gene
(DNA vaccine) product in host cells may not be the
same as that found in the native antigen.
 Present status of DNA vaccines
Since 1990, several groups of workers world-over
have been trying to develop DNA vaccines against
various diseases in experimental animals. Genetic
immunization has been done against a number of
pathogenic organisms. These include influenza A
virus, rabies virus, hepatitis B virus, bovine herpes
virus, HIV type I, and Plasmodium species (malarial
parasite). It must be noted that DNA vaccines have
not been tried in humans for obvious reasons. The
most important being the unknown risks of these
foreign DNAs in human subjects.
 RNA Vaccines
Several workers are trying to use RNA
molecules as vaccines. These RNAs can readily
synthesize the antigenic proteins and offer
immunity. But unfortunately, RNAs are less
stable than DNAs. This poses a big problem for
RNA vaccine manufacture and distribution.
Therefore, the progress in the development of
RNA vaccines has been rather slow compared
to DNA vaccines.
 Plants as Edible Subunit Vaccines
Plants serve as a cheap and safe
production systems for subunit
vaccines. The edible vaccines can be
easily ingested by eating plants.
This eliminates the processing and
purification procedures that are
otherwise needed. Transgenic plants
(tomato, potato) have been
developed for expressing antigens
derived from animal viruses (rabies
virus, herpes virus). A selected list of
recombinant vaccines against animal
viruses produced in plants.
 Attenuated Recombinant Vaccines
In the early years of vaccine research, attenuated strains of
some pathogenic organisms were prepared by prolonged
cultivation — weeks, months or even years. Although the
reasons are not known, the infectious organism would lose
its ability to cause disease but retains its capability to act as
an immunizing agent. This type of approach is almost
outdated now.
It is now possible to genetically engineer the organisms
(bacteria or viruses) and use them as live vaccines, and such
vaccines are referred to as attenuated recombinant vaccines.
The genetic manipulations for the production of these
vaccines are broadly of two types:
✓Deletion or modification of virulence genes of pathogenic
organisms.
✓Genetic manipulation of non-pathogenic organisms to carry
and express antigen determinants from pathogenic
organisms.
✓The advantage with attenuated vaccines is that the native
conformation of the immunogenic determinants is
preserved; hence the immune response is substantially
high. This is in contrast to purified antigens which often
elicit poor immunological response.
Some of the important attenuated vaccines
developed by genetic manipulations are:
• Cholera:
• Creating a new strain of V. cholera:
• Potato as a vehicle for cholera vaccine:
• A group workers have developed a gene altered potato
containing attenuated cholera vaccine. These potatoes
when fed to mice induced immunity against cholera.
• Salmonella Species
• Leishmania Species
 Vector Recombinant Vaccines
Some of vectors can be genetically modified and employed as
vaccines against pathogens.
• Vaccinia viruses is basically the vaccine that was originally used
by Jenner for the eradication of smallpox. The molecular biology
of this virus has been clearly worked out. Vaccinia virus contains
a double-stranded DNA (187 kb)
Vaccinia viruses is basically the vaccine that was originally used
by Jenner for the eradication of smallpox. The molecular biology
of this virus has been clearly worked out. Vaccinia virus contains
a double-stranded DNA (187 kb) that encodes about 200
different proteins. The genome of this virus can accommodate
stretches of foreign DNA which can be expressed along with the
viral genes
The vaccinia virus can replicate in the host cell
cytoplasm (of the infected cells) rather than the
nucleus. This is possible since the vaccinia
virus possesses the machinery for DNA
replication, transcription-DNA polymerase, RNA
polymerase etc. The foreign genes inserted into
the vaccinia virus can also be expressed along
with the viral genome. Thus, the foreign DNA is
under the control of the virus and is expressed
independently from the host cell genome.
Production of recombinant vaccinia viruses:
The development of recombinant vaccinia virus is carried out by a
two-step procedure:
Assembly of plasmid insertion vector:
Fresh vaccinia (cow pox) viruses are processed to release their DNAs.
Now genes from hepatitis B virus, herpes simplex virus and influenza
virus are added one after another and inserted into vaccinia virus
genome. These DNA clusters are cloned in E. coli for increasing their
number and to produce plasmid insertion vectors. The plasmid
contains the foreign subunit genes, the natural vaccinia genes,
including the promoter genes. The recombinant plasmids are isolated
and purified and serve as plasmid insertion vectors.
Production of recombinant vaccinia viruses:
✓The animal cells are infected with plasmid insertion vectors and
normal vaccinia viruses. As the viral replication occurs, the
plasmids are taken up to produce recombinant vaccinia viruses.
The plasmid insertion vector incorporates its genes into
vaccinia virus genome at a place that encodes for the enzyme
thymidine kinase (TK).
✓Thus the recombinant viruses have lost their ability to produce
TK. There are two advantages of loss of TK gene. One is that it
is easy to select recombined vaccinia viruses that lack TK gene
and the second is that these viruses are less infectious than the
normal viruses. The recombinant vaccinia viruses, released from
the cultured animal cells, can be successfully used as vaccines.
These live viral vaccines have some advantages over the killed
or subunit vaccines.
 Advantages
✓Authenticated antigens that closely resemble
natural antigens can be produced.
✓The virus can replicate in the host cells. This
enables the amplification of the antigens for
their action on B-lymphocytes and
T-lymphocytes.
✓There is a possibility of vaccinating several
diseases with one recombinant vaccinia virus.
 Disadvantages
✓The most important limitation is the yet
unknown risks of using these vaccines in
humans.
✓There may be serious complications of using
recombinant viral vaccines in
immunosuppressed individuals such as AIDS
patients.
Other viral recombinant vaccines:
Most of the work on the development of live viral
vaccines has been carried out on vaccinia virus.
Other viruses such as adenovirus, poliovirus and
varicella-zoster virus are also being tried as
recombinant vaccines. Scientists are attracted to
develop a recombinant poliovirus as it can be orally
administered. It might take many more years for the
recombinant viral vaccines to become a reality for
human use.
Delivery of Antigens by Bacteria:
It is known that the antigens located on
the surface of a bacterial cell are more
immunogenic than the antigens in the
cytoplasm. Based on this observation,
scientists have developed strategies to
coat the surfaces of non-pathogenic
organisms with antigens of pathogenic
bacteria.
Delivery of Antigens by Bacteria:
Flagellin is a protein present in the fragella (thread like
filaments) of Salmonella. A synthetic oligonucleotide
encoding the epitope of cholera toxin B subunit was
inserted into Salmonella flagellin gene. This epitope
was in fact found on the flagellum surface. These
flagella-engineered bacteria, when administered to
mice, raised antibodies against the cholera toxin B
subunit peptide. It may be possible in future to
incorporate multiple epitopes (2 or 3) into the flagellin
gene to create multivalent bacterial vaccines.
Recombinant vaccines, which are developed using genetic engineering
techniques to introduce genes from a pathogen into a different organism
(often a harmless virus or bacterium), have shown promise in offering
several benefits, including safety and scalability:
✓Complexity of Design: Designing a recombinant vaccine involves understanding the
genetic makeup of the pathogen and identifying specific genes or proteins that can
induce an immune response. This process requires significant research and
development efforts.
✓Production and Manufacturing: Producing recombinant vaccines can be complex and
expensive. Culturing the host organism (like a virus or bacterium) to produce the
desired antigen or protein requires specialized facilities and expertise. Scaling up
production while maintaining consistency and purity poses challenges.
✓Regulatory Approval: Obtaining regulatory approval for recombinant vaccines
involves demonstrating their safety, efficacy, and consistency in large-scale
production. Meeting stringent regulatory requirements can be time-consuming and
costly.
Recombinant vaccines, which are developed using genetic
engineering techniques to introduce genes from a pathogen
into a different organism (often a harmless virus or
bacterium), have shown promise in offering several benefits:
✓Storage and Stability: Some recombinant vaccines may require specific
storage conditions, such as low temperatures or specialized equipment,
which can pose logistical challenges, especially in regions with limited
infrastructure.
✓Public Acceptance: Public perception and acceptance of new vaccine
technologies, including recombinant vaccines, can impact their uptake.
Addressing concerns about safety, efficacy, and the use of genetically
modified organisms is crucial for widespread acceptance.
✓Cost and Accessibility: The development and production costs of
recombinant vaccines can be higher than traditional vaccines. Ensuring
affordability and accessibility, especially in low-resource settings, remains a
challenge.
Recombinant vaccines, which are developed using genetic
engineering techniques to introduce genes from a pathogen into
a different organism (often a harmless virus or bacterium), have
shown promise in offering several benefits:
✓Emerging Variants: Like other vaccines, recombinant vaccines might face
challenges in addressing rapidly evolving pathogens or variants.
Modifications or updates to the vaccine might be necessary to maintain
effectiveness against new strains.
Advancements in recombinant vaccine technology continue to evolve,
aiming to enhance efficacy, safety, scalability, and accessibility. Some
potential advancements include:
✓Targeted Antigen Design: Refining the selection and design of
antigens within recombinant vaccines to elicit stronger and more
specific immune responses. This includes identifying key antigenic
components of pathogens to improve vaccine effectiveness.
✓Multivalent Vaccines: Developing vaccines that target multiple
strains or variants of a pathogen in a single formulation. This
approach can provide broader protection against diverse strains,
reducing the need for frequent updates or booster shots.
✓Virus-like Particle Vaccines: Creating virus-like particles (VLPs) that
mimic the structure of a virus but are non-infectious. VLP-based
vaccines can induce robust immune responses without the risk of
causing disease because they lack genetic material.
 Advancements in recombinant vaccine technology continue to
evolve, aiming to enhance efficacy, safety, scalability, and
accessibility
✓mRNA Technology: Expanding the use of mRNA technology in recombinant
vaccines. mRNA vaccines, like some COVID-19 vaccines, provide instructions
to cells to produce specific proteins, triggering an immune response.
Advancements in mRNA technology could lead to the development of more
vaccines against various diseases.
✓Self-amplifying RNA Vaccines: Developing vaccines using self-amplifying RNA,
which can replicate itself within cells, potentially leading to a stronger and
more sustained immune response with smaller vaccine doses.
✓Improved Delivery Systems:** Innovations in delivery mechanisms, such as
using nanoparticles or microneedle patches, can enhance the stability and
efficiency of vaccine administration, improving accessibility and storage
requirements.
Advancements in recombinant vaccine technology continue to
evolve, aiming to enhance efficacy, safety, scalability, and
accessibility
✓Plant-Based Vaccines: Utilizing plants as bioreactors to produce recombinant
proteins for vaccines. This approach can potentially lower production costs,
increase scalability, and simplify vaccine distribution.
✓Universal Vaccines: Pursuing the development of universal vaccines that
target conserved elements shared among different strains or even different
pathogens within a specific family. These vaccines aim for long-term
protection against multiple variants.
✓Adjuvants and Formulation Enhancements: Incorporating novel adjuvants or
improving vaccine formulations to boost immune responses and prolong the
durability of protection.
✓Personalized Vaccines: Exploring personalized vaccine approaches that
consider an individual’s genetic makeup or immune profile to create tailored
immunizations for specific populations or diseases.
 Conclusion
Despite these challenges, recombinant vaccines hold promise for
combating various infectious diseases and are actively
researched due to their potential for improved safety profiles
and targeted immune responses. Overcoming these obstacles
through continued research, technological advancements, and
streamlined production processes can lead to more effective and
accessible recombinant vaccines in the future.
Continued research, technological innovations, and collaborative
efforts within the scientific community drive advancements in
recombinant vaccine technology. These advancements hold the
potential to revolutionize vaccine development, offering more
effective, safer, and accessible solutions against a wide range of
infectious diseases.