dna vaccine
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Preface
DNA
VACCINE
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Vaccine saves millions of people life. DNA vaccine is advanced Work in vaccine development and research. This new generation vaccine provides promising hope for our future generation. In this work, mainly discussed fundamental ideas about DNA vaccine and also results of my lab work against bacterial pathogens. For this Preparation, I took help from several articles of relevant topics most of which are mentioned in bibliography. I am greatly indebted to all the authors .Many students help me during my lab work time. I am very much thanking to all of them. I hope this book will useful to students, researchers and also all type of readers.
M.Muruganandam,
Zambia.
CONTENTS
Discovery
Function of vaccines
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DNA vaccines
Mode of Administration
Merits of DNA vaccines
Lab Works
Discussion
Future vaccines
Bibliography.
Discovery
Long ago, people tried to develop different prevention
strategies against infectious diseases, even though they didn’t
know anything about causative agents. These are preliminary
4
attempts for vaccinations. Here these are discussed in detail¹¸².
Chinese try to prevent the smallpox a deadly disease
characterized by pus – filled blisters – by exposing uninfected
individuals to matter from smallpox lesions. This process
known as “variolation”, took a variety of forms. One form
consisted of removing pus and fluid from a smallpox lesion
and using a needle to place it under the skin of the person to
be protected. Another method involved peeling scabs from
lesions, drying and grinding them to powder, and letting an
uninfected person inhale this powder.
The third method involved picking up a small amount
of the scab powder with a needle and then using the needle to
place the powder directly into the individual’s veins. Lady
Mary Wortley Montagu, wife of the British Ambassador to
Turkey, observed this third method in the early 1700s and
brought it back to England. Although the effects of variolation
varied, ranging from causing a mild illness in most
individuals to causing death in a few, the mortality and
morbidity rates due to smallpox were certainly lower in
populations that used variolation than in those that did not.
One person who experienced variolation as child in
the late 1700s was Edward Jenner, a young boy who
survived the process and grew up to become a country doctor
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in England. As a country doctor, Jenner noticed a relationship
between the equine disease known as “grease and a bovine
disease known as “Cow pox.” He saw that farmers who
treated horses with grease lesions often saw the development
of cow pox in their cows, complete with blisters similar to
those seen in smallpox infection. Unlike lethal smallpox,
however, the cowpox blisters eventually disappeared leaving
only a small scar at the site of each blister.
At the same time, Jenner was interested when a
milkmaid told him that she could not many people like the
milkmaid – people who milked cows and who did not get
smallpox even when exposed repeatedly. With this in mind,
Jenner undertook a baring experiment in 1796; he infected a
young boy with cowpox in hopes of preventing subsequent
smallpox infection. After allowing the boy to recover fully
from cowpox, Jenner – in an experiment that would be
considered unethical by today’s scientific community –
intentionally infected the boy with smallpox by injecting pus
from a smallpox lesion directly under his skin. As Jenner had
predicted, the boy did not contract smallpox.
Although Jenner wanted to report his first case study in the
transactions of Royal Society of London, his study was
rejected. Despite this Jenner went on to collect 23 case
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histories over next months and published his own book
detailing his observations. The book was called “An inquiry
into the cases and effects of the variolae vaccinae”, a
disease discovered in some of the western counties of
England, particularly Gloucestershire, and known by the name
of The Cow Pox.” It soon became clear that Jenner’s
experiments had paid off, and that intentional infection with
cowpox protected people from much serious infection with
smallpox. As a result, within a few years thousands of people
protected themselves from the deadly smallpox disease by
intentionally infecting themselves with cowpox.
1.Edward jenner’Book(courtesy-commons.wikipedia.org).
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2..Edward jenner tests his hypothesis for develop a Small pox vaccine.(courtesy-personal.psu.edu)
Jenner’s process came to be called “vaccination,”
after “vacca,” the Latin word for cow, and the substance used
to vaccinate was called a “vaccine.” Now, some 200 years
later, we have progressed from a time when vaccination was a
rare event, and Jenner’s theories about vaccination were not
widely accepted, to the late 1900s when vaccines area so
common place that most children receive multiple
vaccinations before they reach their first birthdays.
The result of such widespread vaccination has been a
marked decrease in diseases which once ravaged the world’s
population. An example of this smallpox, once a major cause
of death world-wide, the smallpox virus is now found only in
freezers in high-containment laboratories at the Centers for
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Disease Control and Prevention (CDC) in Atlanta and the
institute for Viral Preparations in Moscow.1, 2.
Dr. Eugine Tragus chair in molecular cardiology was
first to demonstrate genetic immunization in 1992 with the
development of a gene gun, “which provided DNA coated
micro projectiles shot directly into the cells of animals could
provoke an immune response4.
In 1995, the first clinical trials using injections of
DNA to stimulate on immune response against for HIV. Four
other clinical trails using DNA vaccines against influenza,
herpes simplex virus, T cell lymphoma and an additional trial
for HIV were started in 1996. 3
Functions of Vaccines
The immune system functions to identify and attack
foreign substances; specifically mutant cells, foreign bacteria,
fungi and viruses. When attacked by an antigen (a substance
that causes the production of antibodies when introduced
directly into the body) the immune system goes into
immediate action. It first identifies the specific antigen which
has begun an “invasion” and then begins to use various
abilities. The immune system has the ability to distinguish
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between different antigens; the ability to remember how to
identify foreign antigens; the ability to locate and move
defenses against the foreign antigen and the ability replicate
components in the immune system to increase the immune
response to those foreign antigens.
3. T-lymphocyte.(courtesy-daviddarling.info)
There are four cell type components that make up the immune
system. Most commonly known are the “white blood cells”
(T-cells), phagocytes, B-cells and NK-cells. These four
components together (or separately) fight foreign substances
in the body. T-cells are the “master switch” of the immune
response and are responsible for turning on and off the entire
system. Phagocytes are produced in bone marrow and
function by surrounding and ingesting foreign material. B-
cells are responsible for producing antibodies. There are two
types of B-cells: plasma which is short-lived but produces
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antibodies specific for that particular antigen; and memory
cells which are ready to respond to a second or subsequent
infection by the same antigen. This phase of the immune
cascade is where immunity to various diseases is developed.
This phenomenon is the basis for giving a primary vaccine
followed by a booster shot at a later date. NK-cells’ primary
function is to attack cancer cells.
4.Macrophage(courtesy-en.wikipedia.org)
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5. Natural killer cell attack cancer cell...( courtesy- science photo.com)
When the immune system encounters a virulent
bacteria or virus for the first it synthesizes specific agents to
deal with the infracting organism. Once the immune system
has learned to produce a specific antibody it then produces the
proper amount of antibody as needed. If these learning and
production processes occur quickly enough, then the disease
can be overcome. The next time the immune system
encounters an organism which it has previously met, it
“remembers” what to do to combat the infection. The process
becomes much more rapid thus either reducing the severity of
the disease or preventing the disease altogether. The “body” is
said to have “acquired immunity” vaccines induce immunity
by use of the same principle. Vaccines bear some similarities
to the disease agents. A vaccine introduces the body to altered
antigens of a disease to provoke a defensive response. The
body acquires immunity without suffering the disease. The
ideal vaccine should be able to induce production of both
humeral and cell mediated immure response5.
DNA vaccines
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Vaccines have traditionally been used as weapons
against infections diseases. DNA vaccine is one of the novel
and most powerful vaccines in infections diseases. DNA
vaccines are made of a modified form of infections
organism’s DNA. Recently, encouraging results were reported
for DNA vaccines where by DNA coding for the foreign
antigen is directly injected into the animal so that the foreign
antigen is directly produced by the host animal.
The technique that is being ester in humans
involves the direct injection of plasmid-loops of DNA that
contain gems for proteins produced by the organisms being
targeted for immunity. Once injected into host’s muscle
tissue, the DNA is taken up by host cells, which then start
expressing the foreign protein. The protein serves as an
antigen that stimulate an immune responses and protective
immunological memory3.
Some researchers stated that their view about DNA
vaccines first the DNA is isolated from an infections organism
such as a virus or bacterium. This DNA, in highly purified
form, is altered by special techniques to provide eukaryotic
promoters in front of the gene or genes on this DNA. The
promoter region is located at the beginning of a gene. A
promoter is a special DNA nucleotide sequence that forms a
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recognition site for the enzyme required for gene expression-
the synthesis of messenger RNA using the DNA sequence of
the gene as template. This recognition site allows the enzyme
to associate with DNA and to be oriented properly along the
DNA to synthesize messenger RNA.
The messenger RNA (mRNA) will ultimately be
interpreted by protein-synthesizing machinery inside the cell
known as the ribosome. This interpretation of the message i.e.
The interpretation of the nucleotide sequence of mRNA, will
lead to the synthesis of a protein by the ribosome.
According to John.C.Brown6 Since each species of
all organisms have their own kinds or promoter sequences, the
recognition sites for gene expression ie. , the promoters, for
each species of organism are different. Therefore, bacterial
gene promoters are not recognized by the mRNA synthesizing
enzyme in human cells. Consequently, if bacterial DNA were
to be injected into one of our cells, no gene expression from
the injected DNA would occur. Therefore, it is necessary to
provide a eukaryotic promoter at the beginning of this
bacterial gene- of a kind that human genes have in order for
mRNA to be synthesized6.
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However prof. Stanley. A. plotkin reported that there
is possible to develop DNA vaccine is injecting humans with
DNA segments from pathogenic microorganisms that produce
protective proteins after injection; it leads to production of
immune responses
Genomic DNA vaccine
Genetic immunization is going to be a revolution in
vaccines and it is called genomic up the genome of
mycoplasma into small bits and shooting all the genomic bits
into the skin cells of the mice. Each bit of mycoplasma DNA
made a mycoplasma protein, which produced an immune
response in each mouse. This genomic vaccine fools the
immune system into thinking it has been infected by the real
pathogen4.
rDNA vaccine
Another approach is the DNA fragments carried by
using plasmid vector on this basis; a library of gene fragments
was prepared from mycoplasma pulmonis by cloning the
genomic DNA into a plasmid expression vector. Since this
organism has a relatively small genome (about 106 base pairs),
enough of the total DNA protein-coding sequences might be
expressed to induce immunity to the pathogen. Since only a
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small part of the genetic complement of the organism is
expressed and expression is mostly from only a fragment of
genes and not entire functional proteins, pathogenic effects
would be avoided while all the advantages of broad-based
immunity produced by a DNA vaccine would be present
protection against M. pulmonis has been achieved after
immunization with different expression libraries7.The latest
development in DNA vaccine research shows great promises
against parasitic helminthes9.
Plasmid DNA vaccine
The plasmid DNA vaccines are usually circular
plasmids that include a gene encoding the largest antigen (or
antigens) under the transcriptional control of a promoter
region active in human cells. The coding region of the
inserted gene is followed by transcription termination and
polyaderylation sequences.
To permit selection of plasmid-containing Bacteria
during the production process, the plasmid also contains an
antibiotic resistance gene with bacterial origin of replication.
DNA is generally less costly to produce than peptide or
protein vaccines and is chemically stable under a variety of
conditions DNA vaccines are generally administered
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intramuscularly, using either a needle and syringe or needle
free injector.
The immune response to DNA vaccines results from
uptake of plasmids into cells (including dendrites and muscle
cells) where expression of the target antigen gene (or genes)
occurs. The resulting proteins undergo processing as intra
cytoplasmic antigens, producing peptides that bind to class I
MHC molecules. The presentation of these MHC-bound
peptides on the cell surface stimulated CD8T-lymphocyte
responses. Antibody responses to plasmid encoded proteins
are also observed, suggesting that plasmid-encoded protein
antigens reach and stimulate13. Lymphocytes. DNA vaccines
thus mimic viral infection by inducing both cellular and
humoral immune responses8.
The magnitude of these responses is generally modest
when DNA is used alone. Primate studies 12,13,14,15,16,17,18. And
preliminary results of human trial10, 19. That more potent
specific immune responses could be induce by combining
DNA with adjuvant, by boosting with a recombinant viral
rector or protein or by both adjuvant and boosting. Donnelly
et al 9 have shown that a mixture of plasmids can be used to
induce antibody to the influenza hemoglutirn surface protein
and cyto toxic immunity to the viral nucleoprotein and matrix
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protein. The efficacy of DNA immunization has also been
demonstrated for a variety of viral, bacterial, parasitic and
cancer models. These studies together indicate the potential of
this technology for the development of both animal and
human vaccines. DNA immunization holds substantial
promise as an outstanding contribution of Bio-technology to
modern medicine25.
Mode of Administration
There are various methods for deliver DNA
vaccine the important methods are discussed below.
Intradermal particle Bombardment
A more radical method of introducing DNA involves
the bombardment of DNA-coated gold particles when applied
to the skin, these particles produce good immune responses
with much less DNA than required by other routes such as
intramuscular or intradermal needle injection. This technique
works very well.
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Intranasal drops and intramuscular injections
The application of pure DNA solution (as nose drops)
to the nasal membranes, gives better responses. Reported
that the Intramuscular delivery of plasmid DNA in saline
solution also gives good results22, the method of DNA
vaccination, the highest efficacy was achieved after in vivo is
electroporation and gene gun delivery23.
6. Tattoo DNA vaccination. (Courtesy- wired.com)
Tattooing Method
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DNA vaccines delivered by tattooing have been
shown to induce higher specific humoral and cellular immune
responses than intramuscularly injected DNA. The tattooing is
invasive procedure involving a solid skin, wounding both the
epidermis and the upper dermis in the process and causing
cutaneous inflammation followed by healing now 21.
Modified tattooing devices have been used in medical
research for the delivery of various materials to the skin for
different purpose²¹.
Merits of DNA vaccines
• DNA vaccination provides long-lived immune responses,
unlike many component vaccines that require multiple
inoculations to maintain immunity.
• Vaccines for multiple diseases can all be given in a single
inoculation.
• All DNA vaccines can be produced using similar
techniques the ability to use generic production methods
greatly simplifies the vaccine development and
production process24.
20
• Candidate vaccines can be recovered from diseased tissue.
Microbial DNA can be isolated from the tissue of an
infected animal, purified amplified and screened for
vaccine candidates the DNA vaccine provided
significantly better protection against several strains of
influenza compared with current killed-virus vaccines.
• Another potential advantage of the DNA vaccine is that it
may induce presentation of epitopes that more closely
resemble those found in native flu virus compared with
viral proteins produces in E.Coli, yeast or insect cells.
• The DNA vaccines also do not require the use of formalin
(used to inactivate whole virus) which may help preserve
amino acid side chains in key epitopes. DNA vaccines
have induce robust primary and secondary immune
responses in a variety of animal systems.
• Researchers have yet to prove the safety and efficacy of
DNA vaccines in humans several issues remain to be
addressed. These include concerns over the potential for
induction of tolerance; and clinically significant immune
responses against the injected DNA.
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• Another concern is that the injected DNA may randomly
integrate into the genome25.
• DNA vaccines can be manufactured for more easily than
vaccines compared of an inactivated pathogen, sub
cellular fraction or recombinant protein. Since almost all
plasmids can be manufactured in essentially the same
way, substantial economies of scale can be achieved.
• DNA is very stable and resists temperature extremes
consequently; the storage, transport, and distribution of
DNA-based vaccines are more practical and less
expensive.
• DNA vaccine preventive and therapeutic capabilities26.
• In addition to the commercial, there are vaccines research
and development considerations. It is now possible to
change the sequence of an antigenic protein, or to add
heterologous epitopes, by simply introducing mutations to
the plasmid DNA.
• The immunogenicity of the modified protein can be
directly assured after injecting the plasmid DNA. This
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simple method could increase considerably our
understanding of the immune response to antigens20.
Lab work
In our lab experiments, first we have used
various bacterial preparations as vaccines, such as killed ,live
attunated,toxid,whole protein extraction, genomic DNA and
plasmid DNA ,among them plasmid DNA gives better results
.Then plasmid DNA was mutated by using U.V radiation and it
was used as vaccine. In the next experiments plasmid DNA was
digested by using various restriction enzymes either single and
double digestion then these were used as vaccine. All these
experimental results were presented at various conference and
these abstracts were given below in detail.
Aeromoras hydrophila vaccine27.
Aeromonas hydrophila mainly present in drinking
and ground water of various countries. It mainly gives problem
to immuno suppressive patients and normal host Aeromonas
hydrophila, causing bacteraemia, cellulites to human has not got
a main stay treatment till date. Now is to develop a novel
vaccine. In this study, two experiments are carried out. In this
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first experiment, live attenuated vaccine with various adjutants
such as vitamin A, C and E were tested. In this second
experiment, six various types of vaccines such as killed
vaccines, live attenuated vaccine, toxin vaccine, DNA vaccine,
plasmid DNA vaccine and protein vaccine were studied. Albino
rats were used as test animal and the results shows the
maximum humoral response observed in protein and plasmid
DNA vaccine. In this attempt vitamin A act as best adjuvant.
Staphylococcus aureus vaccine 28.
Staphylococcus aureus cause a verity of diseases in
human through toxin production or invasion. The most
common cause of food poisoning is staphilotoxin, this
bacterium grows in improperly stored food, the cooking
process kill them, but the toxins are heat resistant and it also
infect wounds. In this experiment, staphylococcus aureus
mutant strain was produced by using U.V treatment and
plasmid DNA were isolated from all the mutant strains these
DNA was used as vaccine. The same level of vitamin C was
provided to all the treatments for as adjuvant. After fifteen
days inactivated pathogens was injected to all treatments
including control treatment. After that blood samples of the
entire albino rat were taken then antibody levels and
hematological analysis were taken. The maximum immune
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response was observed in 6th minute treatment strain. So, it is
concluded that this 6 minute treatment strain’s plasmid DNA
is suitable for development of S. aureus vaccine.
.
Genetically Engineered DNA vaccine for Typhoid29.
Typhoid fever is one of important public health
problem. It is mainly spread through contaminated drinking
water and food. In this study, Genomic DNA and plasmid
DNA were isolated from Salmonella typhi and it was injected
to different groups of albino rats. The both DNA were
digested separately by enzymes such as Bam H-I and Pst-I
and also individually double digested. The entire digested
DNA was individually injected to various groups of animals.
One control treatment and one killed vaccine treatment also
maintained during this experiment. After one week, again
gave same booster dose then one week later, blood sample
were collected and analyzed. The maximum immune response
was raised in double digested plasmid DNA treatment and
Pst-I digested plasmid DNA treatment. So it is concluded that
these DNA are highly suitable for DNA vaccine preparation
for typhoid.
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New DNA vaccine for traveler’s Diarrhea
Disease31
Virulent E.coli cause mainly diarrhea and vomiting. It
is also main case for urinary tract infection. Still there is no
good vaccine for human infections. In this attempt DNA
vaccine was developed against E.Coli infection. In this work,
killed vaccine, genomic DNA, plasmid DNA, single digested
plasmid DNA and double digested plasmid DNA were used as
vaccine. One control treatment was always maintained based
on these trails plasmid DNA vaccine, Hind-III digested DNA
and Pst-I + Hind-III double digested DNA shows maximum
cellular immune response. So it is concluded that, above said
DNA preparations are highly suitable for E.Coli DNA vaccine
development.
Engineered plasmid DNA vaccine for staphylococcus
aureus34.
Now a days plasmid DNA has wide variety of
applications in vaccine research. Here it is modified and used
as vaccines. First plasmid DNA was isolated from
Staphylococcus aureus and engineered by various restriction
enzymes. In this work, two experiments were carried out. In
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the first experiment, plasma DNA was isolated and digested
individually by five restriction enzymes such as ECOR-I,
Hind-III, Pst-I, Bam H-I and HaeIII (Medox Company,
Chennai). Then digested plasmid DNA was used as vaccine.
In the second experiment, isolated plasmid DNA was double
digested by these enzymes and used as vaccines. Albino rat
was used as test animal in all the treatment. In the first
experiment, maximum immune response was observed in Pst-
I and Hae-III digested treatment. In the second experiment
maximum immune response were observed in ECOR-
I+HindIII and HindIII+BamHI digested treatments. So it is
concluded that, these double digested treatments are highly
suitable for develop plasmid DNA vaccine for
Staphylococcus aureus.
cocktail plasmid vaccine for common food borne
diseases 35
Our previous studies show that, plasmid DNA
vaccine is one of the best vaccines. So in this attempt, try to
develop a common plasmid DNA vaccine for staphylococcus
aureus, salmonella thyphi and Escherichia coli. In this work,
four treatment works was tested and one control treatment
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was also used. In the first treatment, plasmid DNA was
collected from Salmonella typhi, Escherichia coli and
Staphylococcus aureus. These plasmids DNA was mixed well
and delivers through intramuscular injection. In the second
treatment, all the plasmid DNA were digested by Bam H-I
enzyme and mixed well then these digested plasmid DNA was
used as vaccine. In the third treatment, all the plasmids were
digested by Pst-I enzyme and these also mixed well and used
as vaccine. In the fourth treatment, all plasmids are double
digested by Bam H I and Pst-I enzymes and used as vaccines.
The maximum immure response was observed in double
digested treatment compared to other treatments. So it is
concluded that it is best for development a common vaccine
for these diseases.
DNA vaccine for common food Born diseases30
Nearly 250 food Born diseases have been described.
Bacteria are the causative agents of two thirds of food born
disease out breaks. The important bacterial infections agents
are Salmonella typhi, Staphylococcus aureus, Escherichia coli
etc. In the present attempt, Genomic DNA of above said
organism were isolated and mixed well then used as vaccine.
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The DNA was individually digested by Bam H-I and Pst-I –
Enzymes and then mixed all Bam H-I digested fragments and
mixed well all Pst-I digested fragments of all organisms’
DNA and then used as vaccine. Here controls treatment was
also maintained. Albino rat was used as test animal. After one
week of injection, same dose as booster dose were given to all
animals then one week later blood samples were collected and
analyzed. The maximum immune response was observed in
Pst-I digested DNA mixer. So it is concluding that Pst-I
digested mixer DNA acts as best vaccine for these common
food born infections.
Based on these trails, it is concluded that naked plasmid
DNA act as vaccines and their digested part also act as
vaccines .These findings are summarized and discussed in
next chapter.
Discussion
Vaccines have traditionally been used as weapons
against infectious diseases. DNA vaccines are one of the
novel and most powerful vaccines in infectious diseases.
DNA vaccines are made of a modified form of an infection
organism’s DNA. During DNA vaccine development, most of
29
the researchers construct antigenic code of DNA sequence
with in front of a Eukaryotic promoter sequence for gene
expression. However some researchers made the vaccine by
cutting up the DNA and use as vaccine for e.g. The genome of
mycoplasma was cut into small bits and shooting all the
genomic bits into the skin cells of the mice. Each bit of
mycoplasma DNA made a mycoplasma protein, which
produced an immune response in each mouse4.
Prof. Stanley.A.PlotKin stated that injecting humans
with DNA segments from pathogenic microorganisms that
produce protective proteins after injection. In our lab work
Genomic DNA and Plasmid DNA were isolated from
pathogenic bacteria such as salmonella typhi, Escherichia
coli and staphylococcus aureus and DNA were digested with
various restriction enzymes and deliver by intramuscular
injection then immune response were studied Based on these
trials, the following points are concluded.DNA vaccine is
best compared to other vaccines.
• Mutant plasmid DNA gives some times better
immune response.
• Compare to genomic DNA, plasmid DNA gives
better immune response in the bacterial pathogens.
30
• Mixer of various plasmid DNA of pathogenic
bacteria also gives better immune responses
(develop a combined vaccines).
• Sometimes the digested plasmid DNA (single
digestion or double digestion) gives better response
compare to normal plasmid DNA vaccine.
If go to large scale production (or fine tune experiment),
first, DNA fragments will be separated by agrose gel
electrophoresis then the individual fragments efficacy will be
tested. Some times more than one DNA fragment i.e.
combined fragments synergistically act as best vaccines. So
the correct fragments must be identified than it will be
sequenced and produced more amounts . After preparation of
vaccine, we must go to selection of suitable adjuvant for boost
up the efficacy of vaccine. In future, I hope, we can develop
DNA vaccines for most of the dangerous diseases.
Future vaccine
31
In future, vaccine development will be flourishing to
many directions. Prof. Stanley. A. Plotkin shows his ten
predictions regarding the future vaccines such as vaccine for
multiple diseases, special vaccines for children and old age
people, etc. will be evolve in future. The professor’s
predictions are as given below in details33.
Combined Vaccines
Containing multiple valance will increase valance is
the number of different antigens in vaccine – a trivalent
vaccine has three antigens for e.g. It need fewer injections.
Immunologic adjuvant will come into use in infancy
Immunological adjuvant substances that enhance
responses to vaccination. Although many vaccines are
administered to infants under the age of one year Protection is
slow to develop because of the immaturity of the immure
system. In fact, immunity may fade later in childhood if no
booster doses are given. So we give importance to
immunologic adjutants.
Improve the efficacy of vaccines in an increasingly aged
population
32
The elderly suffer a natural aging of the immune system,
both with respect to antibody production and cellular responses
to infection or vaccination. Here again, we are beginning to
understand the defects that come with age, and correction of
these defects should improve the efficacy of vaccines in an
increasingly aged population.
DNA vaccine
Two new strategies have been come wide spread for
experimental vaccine development.
• Injecting humans with DNA segments from
pathogenic micro organisms that produce protective
proteins after injection and.
• Inserting genes from pathogens into harmless micro
organisms that act as carriers or vectors, for
production of immune responses. Although each
strategy separately may generate useful vaccines, the
combination of the two in a so-called “prime boost
sequence”. Provides synergy.
Thus, there will be vaccinations consisting of prime –
boost. Regimens, particularly in those cases, where
antibodies are in efficient to give complete protection.
33
Vaccination – Immunization using Patches, micro
needles, needle free, etc.
Intramuscular or subcutaneous injections have served
us well us the means to introduce vaccines into humans.
However, there is limitation to the feasibility of
numerous injections and the practical reasons for preferring
other routes of immunization. Thus, intranasal, aerosol
spraying and oral routes of administration are being
intensively explored for creation vaccines. Moreover
transcutanoeus immunization using patches, micro needles
and other ingenious technologies to pass vaccines through the
skin is promising.
Malaria, HIV and tuberculosis are major targets of
vaccine development.
Short-term protection against malaria has already
been achieved. It will need extension of protection. I
respect that regular booster will be necessary to maintain
protection.
BCG vaccine.
34
The Bacillus calmette – Guerin vaccine development
at the Institute Pasteur in lille, France, in the early 20th century
is effective in children but does not present the infection in
adults.
Insertion of genes that code for additional protective
proteins should improve BCG.
HIV has proven to be a difficult target for vaccination.
But a vaccine that reduces the seriousness of infection
and prolongs life, even will not preventing the disease
completely, is likely to be the protect of current clinical trials.
The development of a vaccine that prevents infection entirely
is less likely in the near future.
Influenza remains a banal with deadly infection.
Although the vaccines we have are very beneficial,
better protection will be derived from the inclusion of more
influenza proteins adjuvant and the combined use of live and
killed vaccine33.
The vaccination process may also change in future
one of the important type of future vaccination is needle free
vaccination. Needle free vaccinations are vaccination which is
35
given without the use of a needle. There are a number of
delivery options for needle free vaccinations, ranging from
nasal sprays to patches worn on the skin. The main reason for
the development of needle free vaccination is less painful.
In the developing world, needle free vaccination
would be a huge boon. Needle – free vaccinations would also
be very easy to deliver, encouraging a wider coverage of the
population. One way to deliver needle – free vaccinations is
through mucosal surfaces like the inside of the nose, mouth
and eyes. Vaccines could be smeared directly onto the surface
for absorption or they could be delivered in the form of an
aerosol spray. Oral vaccines can be delivered in droplet from
directly onto the tongue, as has been done historically with the
oral vaccine for polio. Drug companies have also developed
a instrument so called “jet injectors” which force a liquid
vaccine through the pores of the body which is not requiring a
36
needly32
.
7.Needle free injection. (Courtesy the lake wood coop.com)
Bibliography
1. www.st.edu
2. www.macrobiotech.net
3. http://virology-online.com
4. Sean Henahan – Genomic vaccines: A new way to prevent
Diseases – Access. Excellence @ the national health museum,
what’s news.
37
5. Mohanta.K.N and S.K. Majhi (2003), On Fish vaccination
fishing chimes. Vol.23.No.7, PP.39 – 41.
6. John.C. Brown (2002), ‘What the heck is a DNA vaccine’
Jack’s Bugs in the News-Net reference.
7. Barry.M.A., Lai.W.C, Jhonston.S.A (1995) ‘Protection
against mycoplasma infection using expression library
immunization’ Nature; 377: 632 – 5.
8. Laurence Peiperi (2008), Class: DNA plasmid vaccines.
HVTN-vaccines in development-HIV vaccines Trial net works.
PP. 1 – 9.
9. SCHI Stosom Iasis vaccine, - Net reference.
10. Donnelly.J.J, Friedman.A, Martinez D, Montgomery D.L.,
Shiver J.W, Motzed. SL, (1995) preclinical Efficacy of
prototype DNA vaccine enhanced protection against
antigenic drift influenza virus. Nature medicine, 1:583-7.
11. Emini.E (2002) A potential HIV-1-vaccine using a
replication Defective Adenoviral vaccine vector (plenary). 9th
conference on Retroviruses and opportunistic infections
38
seattle, Washington, May-24-28,208, Abstract L5(web cast
presentation available at http:/63/26.3./84/2002/frame.htm)
12. Amara. R.R, Villinger.F, Altman, J.D, Lydy.S.L. O’Neil.SP,
Staprans. S.I, Monte fiori D./C., Xuy, Herndon, JG, Wayatt.LS,
candido. M.A. Kozyrn.L Earl.PL, Smith.JM, Mahl, Grimm.DD,
Hulssy, ML, Miller.J, Mcclure HM, Mcnicholl.JM, Moss. B,
Robinson HL., (2001), control of mucosal challenge and
prevention of AIDS by a multiprotein DNA/MVA vaccine,
science. Apr.6;292 (5514) : 69-74.
13. Borouch DH, Santra S, Schmitz JE, Kuroda MJ., Fu TM,
Wagner. W., Bilska M, Craiu A, Zheng XX, Krivulka GR,
Beaudry.K., Lifton MA, Nickerson CE, Trigora WL, Punt K,
Freed DC, Lewis MG, E Mimi E.A, Shirer JN, Letvin NL (2000)
control of viremia and prevention of clinical AIDS in rhesus
monkeys by cytokine – augmented DNA vaccination science
Oct.20;290 (2491) : 486-92.
14. Shiver JW, Fu T m, Chen.L., Casimiro Dr, Davies ME,
Evants RK, Zhang ZQ, Simon AJ, Trigona WL, Dubey SA, Huang
L, Harris VA, Long RS, Liang X, Handt.L Schleif WA, Zhu L,
Freed DC, Persaud NV , Guan L, Punt KS, Tang A, Chen M,
Willson KA, Collins KB, Heidecker GJ, Ferrandex VR, Perry HC,
39
Joyce JG, Mach. H., Troutman RD, Isopi La, Williams DM,
XU.Z, Bohannon KE, VAlikin DB, Montefirori. DC, Miura A,
Krivulka GR, Lifton MA, Kuroda MJ, Scihmitz JE, Letvin NL,
Caulfield MJ, Bett AJ, Youil.R, Kaslow DC, Emini EA, (2002)
Replication in competent, Adenoviral vaccine vector elicits
effective anti – immunodeficiency-virus immunity. Nature.
Jan.17; 415 (6869): 331 – 5.
15. Kent SJ, Zhao A, Best SJ, Chandler JD. Boyle DB, Ramshow
IA (1998) Enhanced T-Cell immunogenicity and protective
efficacy of a human immune deficiency virus type-1-vaccine
regimen consisting of consecutive priming with DNA and
boosting with recombinant foul pox virus . J. Virol., Dec72
(12): 101: 88-8.
16. Amara RR, Villinger F, Staprans. SI, Altman JD, Monte Fiori
DC, Kozyr NL, Xuy, Wyatt LS, Earl PL, Herndon JG, Mcclure
HM, Moss. B, Robinson HL (2002) Different patterns of
immune responses but similar control of a simian – Human
immune deficiency virus 89.6P Mucosal challenge by
modified vaccine virus Ankara (MVA) and DNA / MVA
vaccines. J. Viral. Aug; 76 (15) : 7625-31.
40
17. Wee EG, Patel S, Mcmichael AJ, Hanke T. (2002) A DNA /
MVA – based canditate human immune deficiency virus
vaccines for kenya induses multi specific T cell responses in
rhesus macagues J. Gen. Virol. Jan; 83 (Pt-1) : 75-80.
18. Robinson HL, Monte fiori DC, Johnson RP. Manson KH,
Kalish ML, Lifson JD, Rizvita Lu S. Hu SL, Mazzara GP, Panicali
DL, Herndon JG, Glickman R, Candido MA, Lydy SL, Wy, and
MS. Mcclure HM, (1999), Neutralizing antibody –
independent containment of immune deficiency virus
challenges by DNA priming and recombinant pox virus
booster immunizations. Wat. Med. July; 5(5): 526-34.
19. Hanke T, Mc Michael AJ, Mwau M, Wee EG, Ceberj I,
Patel. S, Sulton J, Tomlinson.M, Samuel RV, (2002)
Development of a DNA –MVA/ HIVA vaccine for Kenya :
vaccine . July.6;20 (15) :1995-.8
20. Whalen.R.G (1996), DNA vaccines for emerging infections
diseases : what if?. Emerging infectious Diseases . Vol:2
No=3. P :1-15 Net reference.
21. Gopce. NV, Cwy, Olson.G, Warbrifton. AR, Miller BJ,
Couch LH, Wamer WG, Howard PC: (2006) Response of
41
mouse skin to tattooing: use of SKH 1 mice as a surrogate
modle for human tattooing. Toxi cological. Appl. Pharmac
Cal., 209(2): 145 -158.
22. Wolff. JA, Maione RW, Williams P, Chaong. W, A. CsadiG,
Jani A, Felgner PL : (1990). Direct gene transfer into mouse
muscle in vivo. Science 247 (49 49 Pt-1) :1465-1468.
23. Wells DJ, (2004). Gene therapy progress and prospects:
electroporation and other physical methods. Gene ther .
11(18):1363-1369.
24. Sean Henahan, DNA vaccine outlook, Artica, Access
Excellence @ the national health museum what’s news.
25. Sean henchman – DNA provides better flu vaccine; Artica,
Access Excellence @ the national health museum, what
news.
26. DNA vaccines – Inovio Technology – Inovia Bio Medical
Corporation (Net. Reference).
27. Merina Sara Mathew, Thripty mary Eapen, Dinesh. G and
Muruganandam.M. (2008), Evaluation of various vaccines
efficacy for Aeromonas Hydrophila infection “Medico
42
venturus – 08’-National symposium on Emerging Trends in
medical Bio technology symposium proceedings – 6th and 7th
March. Pp:9.
28. Muruganandam.M and Kochet George Ray (2009), A DNA
vaccine for “Staphylococcous awreus” “VERVE-09”
International conference on impact of microbial
Biotechnology on Indian Industry 3rd and 4th Feb Souvenir
.PP.58.
29. Muruganandam.M, Itoop vargeese, Jinu Joseph,
Johnsolamon, and Mahesh.S Kini (2009) Genetically
Engineered DNA vaccine for Typhoid, Chemzen – 09 – one
day National level Technical symposium 17th Sep Souvenir
PP:23.
30. Muruganandam.M, Johnsolamon, Ayyappan, Kumar, John
and Sathesh (2009), DNA vaccine for common food borne
disease , Chemzen- 09 – one day National level Technical
Symposium on 17th Sep. Souvenir PP:24.
31. Muruganandam.M, John Paul, Satesh, Ayyappan, and
Kumar (2009), New DNA vaccines for Traveler’s Diarrhea
43
Disease. Chemzen – 09 one day National level Technical
Symposium on 17th Sep. Souvenir. PP.27.
32. Smith.SE, wise Geek.com
33. Stan lay A. Plotkin (2007) vaccines in the 21st century
SPAN-Nov-Dec. PP: 488-49.
34. Muruganandam.M. (2010), Engineered plasmid DNA
vaccine for Staphylococcus aureus , National conference on
recent-trends and applications of Nanotechnology in
pharmacy and Biology 21st and 22nd Souvenir Jan.PP:54.
Organized by Dept of Pharmacology, Anna University,Trichy.
35.Rajesh.G and M.Muruganandam(2010) Cocktail plasmid
DNA vaccine for common water borne diseases-National
conference on Innovations in Biotechnology and Clinical
research, organized by Department of
Biotechnology,Adhiyaman college of Engg,Hosur-held on 11
th -12 th March 2010.
44
About the author
Prof. M.Muruganandam is working as COE in DMI
–St.Eugene University,Lusaka,Zambia. He has published
more than 50 papers in National and International
journals.
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