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RECOMBINANT

VACCINE
INTRODUCTION

Definition
 Vaccine may lived or weakened strains of
viruses or bacteria which intentionally give
rise to in apparent to trivial infections
 Vaccines may also be killed or inactivated
organisms or purified product derived from
them
INTRODUCTION
 Vaccines involves deliberate exposure to
antigen under condition where disease
should not result.

 It is used to induced protective immunity

 It
should induce antibody formation in the
body
TYPES OF VACCINES
 A. Traditional vaccines
(i) inactive (killed) vaccine
(ii) attenuated vaccine
(iii) Subunit vaccine

 B. New generation vaccines


(i) conjugate vaccine
(ii) Recombinant vaccine
(iii) DNA vaccine
INACTIVE VACCINES

 These are the type of vaccines which are


prepared by micro organisms which are
previously virulent that have been killed
(inactivated) with heat or chemicals
(formaldehyde) so that it can not produced
disease in person who receives the vaccines
Eg. Original Salk vaccine
ATTENUATED VACCINES

 These are lives micro-organs that have been


cultivated under condition which disable their
virulent properties
 They typically provoke more durable
immunological response & are preferred type
for healthy adults
 E.g.. Yellow fever ,measles, mumps
SUBUNIT VACCINE

 Rather than introducing a whole inactivated


or attenuated micro-organism to an immune
system, a fragment of it can create an
immune response
 e.g. is the subunit vaccines against HBV that
is composed of only the surface proteins of
the virus
CONJUGATE VACCINE

 Conjugate vaccines have been developed


to pathogens whose polysaccharide capsules
protect them from phagocytosis eg. S.
pneumoniae, and Neisseria meningitidis
 This polysaccharides outer coats is poorly
immunogenic, by linking these outer coats to
proteins ,the immune system can led to
recognize the as if it were a proteins antigen
RECOMBINANT VACCINE

 Recombinant vaccines are those in which


genes for desired antigens are inserted into a
vector, usually a virus, that has a very low
virulence property.

 The vector expressing the antigen may be


used as the vaccine, or the antigen may be
purified and injected as a subunit vaccine
DNA VACCINE
 DNA vaccines are the newest vaccines and are still
experimental ,although to date they seem to be very
effective and their safely record is good.,
 DNA of pathogen is inserted in animal cell or human
cell. some muscles cells express the pathogen DNA
to stimulate the immune system
 Both humeral and cellular immunity have been
induced by DNA vaccines.
WHAT IS RECOMBINANT TEC.

 This technique was develop in 1980


 This is technique by which desired gene can
be cloned, DNA can be re-sequenced, &
recombinant proteins can be produce
 variety of important molecules for life can be
prepared by introducing desire gene in DNA.
 This technology use for production of
hormones
HOW DOSE IT WORK?

 Each cell of living organism contains DNA,


which contains genetic information
 This DNA(genes) is necessary for the
product of a verity of essential molecules in
the body e.g.. Proteins , fats,
 It was discovered that gene could be
transferred from one cell to another
 Thismean that cell could be reprogrammed to
secrete a particular protein for eg.growth
hormone by introducing gene coding for growth
hormone

 These modified cell – now call as recombinant


cell- transmit new characteristics in daughter cell
Transfer and Cloning of the Insulin Gene

:
Recombinant vaccine

 A recombinant vaccine contains either a


protein or a gene encoding a protein of a
pathogen origin that is immunogenic and
critical to the pathogen function;
 the vaccine is produced using recombinant
DNA technology.
 The vaccines based on recombinant proteins
are also called subunit vaccines.
 Proteins are generally immunogenic, and many
of them are critical for the pathogenic organism.

 The genes encoding such proteins can be


identified and isolated from a pathogen and
expressed in E. coli or some other suitable host
for a mass production of the proteins
Transferring a Gene from one cell to
another
TYPES OF RECOMBINANT VACCINE

 Subunitvaccine
 Gene deleted vaccine
 Vectored virus vaccine
 Other
Subunit vaccine
 A subunit vaccine contains only the proteins which
stimulate the immune system to attack.
 By manipulating DNA that codes for these most
stimulatory proteins, we can mass produce a purified
solution and immunize with only the antigen we want
and no extraneous antigens.
 Eg. The Genitival brand of Feline Leukemia Vaccine
(not currently on the market) is an example of a
subunit vaccine
Gene deleted vaccine

 Gene which is responsible for production of


antigenic response is deleted or removed from
pathogen ,

 Pathogen is then inserted in to the body

 Due to removal of antigenic gene could not able to


produce infection & physical illness
Vectored virus vaccine
 The DNA for the stimulatory proteins are inserted into harmless
viruses. The live harmless virus is able to provide a very natural
immune stimulation and will express the stimulatory proteins native
to a harmful virus. In this method, one gets the benefits of the
modified live vaccine with the benefits of the subunit vaccine. Eg.
Rabies vaccine

 This technology is currently in use to vaccinate wildlife against


rabies The new PUREVAX rabies vaccine for cats is also a
vectored virus vaccine.
Other vaccine
 the "naked DNA or RNA" vaccine. Here, DNA from
the infectious agent is injected into the host. No
proteins. No putting the DNA inside any viruses. Just
plain raw DNA.
 This DNA is taken up by the local muscle cells and
soon the proteins coded for by this DNA are being
produced. The immune system thus receives its
stimulation without risking exposure to an infectious
agent of any kind.
METHOD of PREPRATION
 The first step is to identify a protein that is both
immunogenic and critical for the pathogen.

 The gene encoding this protein is then identified and


isolated.

 The gene is integrated into a suitable expression


vector and introduced into a suitable host where it
expresses the protein in large quantities.
 The protein is then isolated and purified from the host cells

 It is used for the preparation of vaccine.

 The host organisms used for expression of immunogenic


proteins to be used as vaccines may be anyone of the
following.
1. A genetically engineered microorganism, e.g., yeast for the
expression of hepatitis B surface antigen (HBsAg) used as
vaccine against hepatitis B virus (approved for marketing in India).

2. Cultured animal cells, e.g., HBsAg expressed in CHO (Chinese


hamster ovary) cell line and C-127 cell line.

3. Transgenic plants, e.g., HBsAg, HIV-l (human immunodeficiency


virus-I) epitope (in experimental stages).

4. Insect larvae; the gene is integrated into a bacculovirus genome,


which is used to infect insect larvae. Often a very high quantity of
the recombinant protein is produced.
Recombinant vector vaccine
 Genes encoding for major antigens of especially
virulent pathogens can be introduced into attenuated
viruses or bacteria.
 The attenuated organism serves as a vector, which
replicates within the host and expresses the gene
product of the pathogen.
 Various organisms have been utilized for vector
vaccines and include adenoviruses, attenuated
poliovirus, vaccinia virus, BCG strain of
Mycobacterium bovis and attenuated strains of
Saimonel/a.
 vaccine virus, which has been used to
eradicate smallpox is widely employed as a
vector vaccine. This large and complex virus
containing a genome of about 200 genes,
can be manipulated to carry scores of foreign
genes without any impairment in its capacity
to infect host cells and replicates.
HBV recombinant vaccine

 Description-
Hepatitis B Vaccine (Recombinant) is a non-
infectious subunit viral vaccine derived from hepatitis
B surface antigen (HBsAg) produced in yeast cells. A
portion of the hepatitis B virus gene, coding for
HBsAg, is cloned into yeast, and the vaccine for
hepatitis B is produced from cultures of this
recombinant yeast strain according to methods
developed in the Merck Research Laboratories.
 Vaccine must contain antibody against surface
antigen(HBV Ag)
 It was first made from the serum of people who had
been HBV infected and subsequently cleared the
infection. Because these individuals naturally
produced anti-HBsAg IgG and then cleared the
infection, they were immune to reinfection with HBV.
 The serum from these individuals was taken and
purified antibody was passively administered to
others in order to induce immunity
Recombinant tec.

 In an infected individual HBsAg is present in


two forms:
 as a protein on the surface of the 42 nm
virion (also called the Dane particle)
 and as a secreted 22 nm particle that is a
hollow sphere of surface antigen.
 gene encoding for this proteins is
determine and is inserted in to the vector
 Two types of vector is used

1. Yeast : sacchromyces cervices

3. Chinese hamster overy


Yeast cell

 yeast assembled this 22 nm protein(s gene)


in the same way that surface antigen
assembles and is secreted in humans.
 Therefore, the artificial surface antigen
resembled the naturally occuring particle.
Chinese hamster ovary (CHO)

 the recombinant plasmid was transfected into


Chinese hamster ovary (CHO) cells.
 this plasmid was different than the
recombinant yeast plasmid because it
included an extra piece of the genome While
the yeast plasmid included only the S gene of
HBV, the CHO clones used the S gene plus
the pre-S2 portion of the genome,
 creating a vector that began one start codon
upstream in the genome.

 In this case as well, the 22-nm particle


containing the immunogenic HBsAg was
produced by the artificial system as that of
yeast .
 Data from one of many efficacy trials...
image taken from Da Villa et al. Res Virol. 1998. 9.
Production of vaccinia vector vaccine

 The gene that encodes the desired antigen (organ) is


inserted into a plasmid vector adjacent to a vaccine
promoter

 When this vector is incubated in tissue culture , the


antigen & promoter inserted in to the vaccinia virous
genome