1

Preface

DNA

VACCINE

2

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

3

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

5

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

6

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).

7

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

8

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

9

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

10

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)

11

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

12

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

13

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.

14

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

15

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

16

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

17

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.

18

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

19

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.

21

• 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

22

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

23

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

24

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

26

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

27

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.

28

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)

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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

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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.