Chapter 7

Chapter 7
Vaccines
UNIVERSITY OF BALAMAND
FACULTY OF SCIENCES
DEPARTMENT OF BIOLOGY
BIOL 287
BIOTECHNOLOGY AND RECOMBINANT DNA
Takla El Khoury, Ph.D.

Molecular and Cell Biology
Biology Department
Faculty of Sciences
Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

of Recombinant DNA, Fifth Edition American Society for Microbiology
Bernard R. Glick, Cheryl L. Patten 1752 N St. NW, Washington, DC 20036-2904
Chapter 7
Vaccines -Vaccination
➢200 years ago, in 1796, Edward Jenner English doctor had discovered the principle of
vaccination.
➢Today, millions of people are alive and well due to the more than 70 vaccines that are
now licensed for use against a wide range of pathogenic agents.
➢The causative agents of diphtheria, polio, rubella and others gave rise to more than 39
million infections in the 20th century in Canada and the United States.
➢Vaccines have since reduced considerably the frequencies of these diseases (typically
by 1000-fold or more).
Table 7.1

Chapter 7
Vaccines -Vaccination
➢The Global Alliance for Vaccines and Immunization estimates that more than 1.5
million children die every year from vaccine-preventable diseases.
➢Effective vaccines are also not yet available for many leading diseases, including human
immunodeficiency virus (HIV)/AIDS, tuberculosis, and malaria, which produce more
than 5 million deaths globally each year
Table 7.2

Chapter 7
Vaccines -Current and Future Vaccines
Figure 7.1
➢Current vaccines typically consist of either a

killed (inactivated) (Fig A) or a live/nonvirulent
(attenuated) (Fig B) form of an infectious agent
➢ Traditionally, the infectious agent is grown in

culture, purified, and either inactivated or
attenuated without losing the ability to evoke an
immune response that is effective against the
virulent form of the pathogen.
➢Notwithstanding the considerable success that

has been achieved in creating effective vaccines
against diseases such as German measles,
diphtheria, whooping cough, tetanus, smallpox,
and poliomyelitis

Chapter 7
Vaccines -Current and Future Vaccines
➢There are a number of limitations to the current mode of vaccine production
•Not all infectious agents can be grown in culture, so no vaccines have been developed
for a number of diseases.
•Production of animal and human viruses requires animal cell culture, which is
expensive.
•Both the yield and rate of production of animal and human viruses in culture are often
quite low, making vaccine production costly.
•Extensive safety precautions are necessary to ensure that laboratory and production
personnel are not exposed to a pathogenic agent.
•Batches of vaccine may not be killed or may be insufficiently attenuated during the
production process, thereby introducing virulent organisms into the vaccine and by
mistake spreading the disease.
•Attenuated strains may revert, a possibility that requires continual testing to ensure that
the reacquisition of virulence has not occurred.
•Not all diseases are preventable through the use of traditional vaccines.
•Most current vaccines have a limited shelf life and often require refrigeration to
maintain potency. This requirement creates storage problems in countries with large
unelectrified rural areas.
Chapter 7
Vaccines -Subunit Vaccines
➢Traditional vaccines generally consist of either killed or attenuated forms of the

pathogenic agent. The antibodies elicited by these vaccines initiate an immune response
to inactivate (neutralize) pathogenic organisms by binding to proteins on the outer
surface of the agent. So, do vaccines need to contain the whole organism, or will
specific portions of pathogenic organisms be sufficent?
➢For disease-causing viruses, purified outer surface viral proteins, either capsid or
envelope proteins, are often sufficient for eliciting neutralizing antibodies in the host
organism. Vaccines that use components of a pathogenic organism rather than the
whole organism are called "subunit" vaccines
Figure 7.2

Chapter 7
Vaccines - advantages and disadvantages to the use of subunit Vaccines
➢On the positive side, using a purified protein(s) as an immunogen ensures that the
preparation is stable and safe, is precisely defined chemically, and is free of extraneous
proteins and nucleic acids that can initiate undesirable side effects in the host organism.
➢On the negative side, purification of a specific protein can be costly, and in certain
instances, an isolated protein may not have the same conformation as it does in situ
(within the viral capsid or envelope),with the result that its antigenicity is decreased.
➢Obviously, the decision to produce a subunit vaccine depends on an assessment of

both biological and economic factors.
Figure 7.2

Chapter 7
Vaccines - Cholera
➢Cholera can lead to death within 24 hours if left untreated. Without treatment, severe
infection by Vibrio cholera, the causative agent of cholera, has a mortality rate of 30%
to 50%.
➢The bacterium V.cholerae colonizes the small intestine and secretes large amounts of
the pathogenic hexameric enterotoxin.
➢This protein consists of one A subunit, that has ADP (adenosine diphosphate)
ribosylation activity and stimulates host adenylate cyclase, and five identical B
subunits that bind specifically to an intestinal mucosal cell receptor.
➢The A subunit has two functional domains: the A1 peptide, which contains the toxic
activity, and the A2 peptide, which (noncovalently) joins the A subunit to the B
subunits.
Figure 7.5

Chapter 7
Vaccines - Cholera
➢Until few years ago, a traditional cholera vaccine consisting of phenol-killed

V.cholerae was in common use. This vaccine generated only moderate protection,
typically lasting from about 3 to 6 months
➢More recently, a vaccine (Dukoral) consisting of heat-inactivated V.cholerae Inaba

classic strain, heat-inactivated Ogawa classic strain, formalin-inactivated Inaba El Tor
strain, formalin-inactivated Ogawa classic strain, and a recombinant cholera toxin B
subunit, has come into use.
➢This newer vaccine is taken orally (two doses 1week apart), and an additional booster
immunization is not required for about 2 to 3 years.
Figure 7.5

Chapter 7
Vaccines - SARS
Figure 7.6
➢The first new infectious disease of the 21st century was identified as Severe Acute
Respiratory Syndrome, or SARS
➢The SARS coronavirus (SARS-CoV) is an enveloped virus that contains a single

stranded plus-sense RNA genome of ~30 kb. The viral spike protein, which is inserted
into the viral membrane, binds to a receptor protein that is present on the surfaces of
mammalian host cells.
➢The spikes of SARS-CoV consist of trimers of an S protein that is composed of two

subunits, S1 containing the receptor binding domain and the S2 subunits that mediate
fusion between the viral and host cell membranes.
Chapter 7
Vaccines - SARS
Figure 7.6
➢After the virus enters the body, it binds via the S1 subunit to the receptor, the
angiotensinconverting enzyme 2 (ACE2), including pneumocytes in the respiratory
system.
➢Following the binding of the virus, the viral and cell membranes fuse, facilitating the
entry of virus into the cell.
➢Thus, the extracellular domain of the S1 subunit is an attractive candidate for the
development of a subunit vaccine to elicit antibodies that block the interaction between
virus and host cell.
Chapter 7
Vaccines - Human Papillomavirus
➢Human papillomavirus (HPV) is a non-enveloped virus that is sexually transmitted.
While most infections are benign and often asymptomatic, persistent infection with some
strains of human papillomavirus is associated with the development of cervical and
related cancers, as well as genital warts.
➢Over 100 different types of HPV exist, and 15 types have carcinogenic potential. Most
genital warts are caused by HPV-6 and HPV-11,while HPV-16 and HPV-18 account for
about 70% of invasive cervical cancers.
➢Thus, a vaccine that prevents HPV-16 and HPV-18 infection could significantly reduce
the incidence of cervical cancer.
➢It previously shown that the viral major capsid protein L1 can selfassembled into
virus-like particles that resemble papillomavirus virions, and these particles are highly
immunogenic, inducing neutralizing antibodies directed against the whole live virus.
➢The gene for the L1 protein from each of four virus types was cloned and expressed
in a recombinant Saccharomyces cerevisiae (yeast) strain. Following separate
fermentations of each of the four yeast strains, the viral capsid proteins assembled into
virus-like particles (the viral capsid without any other viral proteins or the viral nucleic
acid).
Chapter 7
Vaccines - Human Papillomavirus
Figure 7.9
➢ These virus-like particles were then purified and combined to form the quadrivalent
vaccine
➢In June 2006,the US. FDA approved the vaccine, called Gardasil; is quadrivalent,
containing virus-like particles assembled from the L1 proteins of the above-mentioned four
types of human papillomavirus (Fig).In the combined data from the phase II and III studies,
efficacy against external genital lesions from all 4 HPV types was 95% to 99%.
➢In addition to Gardasil, a second HPV vaccine, Cervarix, was developed, and then
approved by the U.S.FDA in 2009. This vaccine is a combination of HPV-16, -18, and -20
L1 proteins that are produced in insect cells in culture.
➢Like Gardasil, this vaccine also requires three injections.

Chapter 7
Vaccines - Vaccine antigens delivery
Figure 7.12
➢ Delivery of antigens by lactic acid bacteria often has the added
benefit of inducing both mucosal and systemic immune responses.
➢ Moreover, many lactic acid bacterial strains have the ability to act
as adjuvants that enhance the immune response induced by the
antigen that they have been engineered to encode.
➢ This is a schematic overview of the development of a lactic acid

bacteria subunit vaccine.
➢ In this procedure, it is essential that the gene encoding the target

antigen be inserted into the bacterial chromosomal DNA rather than
being present on a plasmid.
➢ This ensures that no antibiotic resistance genes, often used as

plasmid selectable markers, will be transferred to other bacteria and
subsequently proliferate in the environment

Chapter 7
➢ Although lactic acid bacterial vaccines have been shown to be effective with
mice, none of these vaccines is currently in clinical trials.
Table 7.3

Chapter 7
Figure 7.13
➢ One recent technique that is still being tested is based on a device called a
Nanopatch.
➢ Traditionally, and still in common use today; a needle and syringe is used to inject a
liquid vaccine into muscle (Fig.A).
➢ However, it has been hypothesized that vaccines might be much more effective if
they were delivered into the narrow layer (dermis) that is just beneath the surface of
the skin (Fig.B).This layer contains a high density of antigen-presenting cells that
are essential to generate a protective immune response.
➢ Intradermal injection can achieve comparable immune responses to intramuscular

injection while using only about one-tenth of the vaccine dose.
Chapter 7
Figure 7.13
➢ As an alternative to intradermal injection, some researchers developed microneedles

to more precisely deliver the vaccine to the skin (Fig.C).
➢ More recently, other scientists developed a Nanopatch technique wherein a very

small and densely packed array (1cm2 square of silicon with ≈20,000
microprojections on its surface) is used lo directly target the vaccine to the dermis
(Fig.D).
➢ Preliminary result with the Nanopatch show that it is up to 10 times more effective
than any other vaccine delivery methods, and this increased efficiency comes
without the reliance on an added adjuvant, and with only a single vaccination
Chapter 7
Vaccines - Peptide Vaccines
Figure 7.14
➢ The question arises as to whether a small discrete portion (domain) of a protein can act
as an effective subunit vaccine and induce the production of neutralizing antibodies.
➢ Intuitively, one might expect that only the portions, or domains, of a protein that are
accessible to antibody binding, that is, those on the exterior surface of the virus, would
be immunologically important and that those located in inaccessible regions inside the
virus particle could be ignored if they do not contribute to the conformation of the
immunogenic domain (Fig.7.14).
Chapter 7
Vaccines - Peptide Vaccines
Figure 7.14
➢ If this argument has validity, it is possible that short peptides that mimic epitopes will
be immunogenic and may be used as peptide vaccines.
➢ This approach requires that an epitope consists of a short stretch of amino acids, which
does not always occur naturally, and the peptide must be able to assume the same
conformation as the epitope in the intact pathogen.
➢ A single epitope may not be sufficiently immunogenic.

Chapter 7
Vaccines - Malaria
Figure 7.15
➢ Malaria is a parasitic disease transmitted to humans and

other vertebrates by mosquitoes and is potentially fatal.
➢ The genus Plasmodium consists of approximately 125

known species of parasitic protozoa, 5 of which
(Plasmodium falciparum, P.vivax, P. ovale, P.malariae,
and P. knowlesi) infect humans and cause malaria.
➢ Because P. falciparum elicits the most severe malaria

disease and the most deaths, it is the target of most
vaccine development efforts
➢ The Plasmodium life cycle is very complex. Sporozoites

from the saliva of a biting female mosquito are
transmitted to either the blood or the lymphatic system
and then migrate to the liver and invade liver cells
(hepatocytes) (Fig).

Chapter 7
Vaccines - Malaria
Figure 7.15 ➢ In hepatocytes, the parasite grows, divides, and
propagates as haploid merozoites before budding off in
merosomes containing hundreds or thousands of
merozoites.
➢ These merosomes lodge in pulmonary capillaries and

invade the red blood cells, where the parasite
proliferates producing new merozoites, which then
leave the red blood cells and travel within the
bloodstream to invade new blood cells.
➢ The parasite eventually forms gametocytes, that develop

gametes. Fusion of the gametes leads to the formation
of new sporozoites in the mosquito that can infect new
individuals, spreading the disease.
➢ In the life cycle of the malaria parasite, it is the asexual

bloodstage multiplication that is responsible for most of
the acute symptoms of the disease.
Chapter 7
Vaccines - Malaria
➢ In areas where malaria is endemic, some individuals show considerable resistance to

the disease, they made antibodies against a parasite protein that prevented the growth
of the parasite.
➢ Following a detailed study, it was determined that the protective antibodies targeted
merozoite surface protein 3. When this protein was examined in different strains of
Plasmodium, it was observed that while the N-terminal part of the protein varied
considerably from one strain to another, the C-terminal end of the protein was
highly conserved among the various isolates of the parasite.
➢ It was therefore decided to chemically synthesize peptides that corresponded to small

portions of the C terminus of merozoite surface protein 3 (Fig).
Figure 7.16

Chapter 7
Vaccines - Malaria
➢ The peptides were tested against serum from individuals who were resistant to the
parasite and the antibodies were affinity purified based upon their interaction with
one or more of the peptides.
➢ The antibodies that bound to the peptides were then tested. Antibodies directed
against peptides B, C, and D had a major inhibitory effect on parasite growth
➢ Therefore, a peptide representing amino acid residues 181 to 276 of merozoite

surface protein 3 was chemically synthesized. This peptide is currently being tested
in clinical trials as a novel malaria vaccine
Figure 7.16

Chapter 7
Vaccines - Malaria
➢ Although a vaccine containing a merozoite antigen would protect against the

erythrocyte stage of infection, a pre-erythrocyte-stage vaccine would block infection
and prevent the parasites from reaching the blood and causing disease.
➢ In this regard, in November 2012, a malaria vaccine that targeted the sporozoite stage
was tested in phase III of clinical trials.
Figure 7.17

Chapter 7
Vaccines - Malaria
➢ The peptide vaccine, RTS,S/AS01,is comprised of the antigen RTS,S and a liposomal
adjuvant, AS01, to boost the immune response.
➢ The antigen is a hybrid of peptides derived from the repeat (R) and T-cell epitope (T)
regions of the immunodominant circumsporozoite protein, which covers the exterior
of the sporozoite and the hepatitis B virus surface antigen (S).
➢ The results of a small phase III clinical trial with children ages 5 to 17 months was
promising, finding 55.8% protection against malaria.
Figure 7.17

Chapter 7
Vaccines - Malaria
Figure 7.18
➢ Some researchers consider using nanoparticles is as both a

delivery system and adjunct to increase the body’s immune
response to a vaccine.
➢ Therefore, researchers used a complex nanoparticle composed

of multilamellar lipid vesicles to deliver a recombinant
Plasmodium vivax circumsporozoite antigen, VMP001,
entrapped in an aqueous core and anchored to lipid bilayer
surfaces
➢ P. vivax is the second most serious malaria-causing parasite

(after P. falciparum) infecting 130 to 390 million people every
year.

Chapter 7
Vaccines - Malaria
Figure 7.18
➢ Immunization with these nanoparticle complexes together with

monophosphoryl lipid A (which stimulates the immune
response) yielded high-titer and high-avidity (affinity for the
antigen) antibody responses in mice against the VMP00l
protein.
➢ In addition, the immune response against this protein lasted for

more than one year at 10-fold lower doses than when
conventional adjuvants were used together with purified
VMP00l protein.
➢ While these data are still preliminary they indicate that the use
of nanoparticles is promising as both a vaccine delivery and
adjuvant strategy.

Chapter 7
Vaccines - Genetic Immunization: DNA Vaccines
➢ DNA vaccination is a relatively new technique for protecting

animals and humans against disease by injecting them with
genetically engineered DNA instead of a protein antigen.
➢ The gene in the injected DNA directs cells to produce a

protein antigen, effectively acting as a subunit vaccine.
➢ An advantage of genetic immunization, besides bypassing

the need for purified protein antigens, is that it triggers a
response against only the protein encoded on the plasmid and
not against the plasmid itself

Chapter 7
Vaccines - Genetic Immunization: DNA Vaccines
Figure 7.19
➢ In one early series of experiments, mice were injected in the quadriceps of both legs
with an E.coli plasmid carrying the cDNA for influenza A virus nucleoprotein under, the
transcriptional control of promoter.
➢ Although the expression of the nucleoprotein was too low to visualize on gels,
nucleoprotein-specific antibodies were observed in the blood of the test mice 2 weeks
after the initial injection.
➢ In comparison to control mice, the nucleoprotein-injected mice were significantly

protected against the lethal effects of influenza virus infection. Moreover, nucleoprotein
-injected mice were also protected against a different strain of influenza
Chapter 7
Vaccines - Delivery: DNA Vaccines
➢ One of the problems with the use of DNA vaccines
in large animals and humans compared to mice is
that the transfection efficiency of introduced plasmid
DNA is often insufficient to generate a protective
immune response.
➢ One approach to deliver foreign DNA to animal cells

Figure 7.20 utilizes biodegradable microscopic (0.3- to l.5-µm)
polymeric particles with a cationic surface that binds
the plasmid DNA (Fig).
➢ Plasmid DNA is bound to the surfaces of these

"microparticles“ and is slowly released over a period
of 2 to 3 weeks after inoculation of an animal.
➢ Using microparticles, it was possible to achieve the

same biological effect as with naked DNA with
about 250-fold less DNA, demonstrating the
potential of this approach.
Chapter 7
➢ In addition the level of antibodies induced by the

expression of plasmid-encoded genes bound to
microparticles was significantly enhanced by:
Figure 7.20
i. addition of vaccine adjuvant aluminum phosphate
ii. the use of nanoparticles 0.05µm in diameter that

were coated with poly-L-lysine.
➢ In contrast to naked DNA, DNA bound to

microparticles induced potent cytotoxic T-
lymphocyte responses at a low dose.

Chapter 7
Figure 7.21
➢ Notwithstanding the lack of immunogenicity of the plasmid vector, the presence of one
or more unmethylated CpG rich oligonucleotide fragments on the plasmid can stimulate
the immune system in inoculated mammalian and aquatic animals.
➢ Importantly, in some instances the inclusion of CpG oligonucleotide motifs may replace
the use of adjuvants (that can often irritate the animal or person being vaccinated) in the
delivery of DNA vaccines.
➢ Synthetic unmethylated CpG oligonucleotides differ from ordinary microbial DNA in

that they typically have a partially or completely phosphorothioated backbone in place
of the usual phosphodiester backbone. They also, often have a poly G tail at the 3' end,
or the 5' end, or both.
Chapter 7
Figure 7.21
➢ The phosphorthioated backbone protects the oligonucleotide from being degraded by

nucleases when the plasmid is introduced into the inoculated animal while the poly G
tail enhances the cellular uptake of the plasmid.
➢ In one experiment, a plasmid vector that was used to inoculate fish with a DNA
vaccine against viral hemorrhagic septicemia was modified to include several CpG
oligonucleotide motifs with the result that the immunogenicity of the vaccine was
significantly increased.
➢ In this experiment, when vectors with zero, two, or four regions of unmethylated CpG
rich oligonucleotide were compared in terms of the immune response against viral
hemorrhagic septicemia, the plasmid that contained four regions of unmethylated CpG
rich oligonucleotide was the most immunogenic.
Chapter 7
➢ DNA vaccines that are designed for delivery to

mucosal surfaces are similar in principle to those
used for intramuscular or intradermal delivery.
➢ To increase plasmid uptake and decrease its

subsequent degradation, various methods of
formulating DNA have been tried
➢ Most likely, the electrical pulse increases the

transfection efficiency of the added DNA, and it
is becoming a method of choice for clinical
administration of certain DNA vaccines.

Chapter 7
Figure 7.22
➢ A modified strain of the invasive (diarrhea-causing) bacterium Shigella ftexneri has

been developed to facilitate the delivery of foreign DNA into animal cells for genetic
immunization (Fig.7.22).
➢ Shigella can enter animal epithelial cells, escaping the phagocytic vacuole, and the
bacterium can direct plasmid DNA to the nucleus of the host cell, where, if the
introduced gene(s) contains a eukaryotic promoter, it is transcribed.
➢ Shigella is normally a human pathogen and as such would not be an acceptable DNA
delivery system. Therefore, to use Shigella, it was first necessary to construct a non
pathogenic version of the wild type organism.
Chapter 7
Figure 7.22
➢ This was done by engineering the bacterium to be toxin deficient (deleting the toxin
gene) and (ii) unable to proliferate in host cells.
➢ The latter was accomplished by making a deletion mutation in the Shigella asd gene,
which encodes the enzyme aspartate ß-semialdehyde dehydrogenase. This enzyme is
normally involved in the synthesis of the bacterial cell wall constituent diaminopimelic
acid; therefore, the mutant strain cannot grow unless diaminopimelic acid is added to
the growth medium.
➢ Shigella strains with the asd mutation can invade animal epithelial cells and deliver
their plasmid DNA; however, once present, the Shigella cells are unable to proliferate
in host cells
Chapter 7
Figure 7.23
➢ To avoid the use of antibiotic resistance marker genes, researchers have developed a
series of minimalistic immunogenically defined gene expression (MIDGE) vectors
➢ Following insertion of the gene of interest into a MIDGE vector, the antibiotic
resistance gene is excised from the vector, and an oligonucleotide cap is added
specifically to both ends of the linearized DNA that contains only the gene of
interest with a promoter/intron and a polyadenylation signal.
➢ The portion of the plasmid containing the antibiotic resistance gene, which is not
protected by the oligonucleotide caps, is degraded by exonucleases.
➢ The capped ends are resistant to exonuclease digestion. The purified, capped linear
MIDGE vector is then used directly for transfection. These vectors have been used
successfully as a substitute for plasmid vectors.
Chapter 7
Figure 7.24
➢At the present time, there is a commercial protein vaccine against hepatitis B virus
that is based on the small hepatitis B surface antigen.
➢Unfortunately, this vaccine does not induce a protective immune response in about
10% of the people who are vaccinated.
➢To remedy this problem, scientists developed DNA vaccines encoding the small or the
large hepatitis B surface antigen.
➢In both instances the antigen genes were spliced into MIDGE vectors (Fig.). The
immunogenicity of these vectors was then assessed in mice and pigs.

Chapter 7
Figure 7.24
➢In both animal models, vectors encoding the small protein induced stronger responses
than vectors encoding the large protein
➢The small protein-specific antibody levels were higher than antibody levels that
would be considered to be protective in humans
➢Although the antibody titer in immunized animals declined by six months after the
final immunization, the decline was less pronounced when the DNA vaccine was used.
➢While these results are still preliminary and this DNA vaccine remains to be tested in
humans, the results are nevertheless encouraging.

Chapter 7
Vaccines - DNA Vaccines:Cancer
Table 7.6

Chapter 7
Vaccines - Attenuated Vaccines: Herpes Simplex Virus
Figure 7.26
➢Given the failure of subunit vaccines directed against

glycoprotein D to protect humans against HSV, scientists
tried another approach to develop a vaccine against this
virus.
➢In this case, a mutant of HSV was engineered with

glycoprotein D deleted. Since glycoprotein D is required for
HSV to enter into cells, deleting it prevented the cell to cell
spread of the virus, marking the attenuated strain
completely safe
➢In order to make an attenuated vaccine against HSV-2,

researchers grew mutant HSV-2 (with the glycoprotein D
gene deleted from its genome) in a cell line that expresses
the HSV-1 version of glycoprotein D.

Chapter 7
Figure 7.27
➢The mutant HSV-2 virus assembled the HSV-1glycoprotein D produced by the cell
into its envelope membrane so that when it was introduced into a mouse, the mutant
HSV-2 with HSV-1 glycoprotein D was able to enter the mouse’s cells.
➢However, upon replication of the mutant HSV-21 the progeny viruses were not able
to infect new cells because the gene encoding glycoprotein D was not present.
➢The HSV-2 surface proteins activated the mouse’s immune system to produce
antibodies against HSV-2.

Chapter 7
Figure 7.27
➢In this case the mutant HSV-2 evoked the immune system's antibody-dependent cell-
mediated cytotoxicity (ADCC) response.
➢In the ADCC response, antibodies recognize HSV-2 infected cells that present the
viral antigens on their surface.
➢Natural killer (NK) cells bind to the Fc portion of the antibodies on the surface of the
infected cells and mediate the killing of the infected cells.
➢Subsequent infection by wild-type HSV-2 will activate this defense response. This
vaccine remains to be tested in humans.
Chapter 7
Vaccines
The End !!


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BIOTECHNOLOGY AND RECOMBINANT DNA

Takla El Khoury, Ph.D.

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢Current vaccines typically consist of either a

➢ Traditionally, the infectious agent is grown in

➢Notwithstanding the considerable success that

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢There are a number of limitations to the current mode of vaccine production

➢Traditional vaccines generally consist of either killed or attenuated forms of the

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢Obviously, the decision to produce a subunit vaccine depends on an assessment of

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢Until few years ago, a traditional cholera vaccine consisting of phenol-killed

➢More recently, a vaccine (Dukoral) consisting of heat-inactivated V.cholerae Inaba

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢The SARS coronavirus (SARS-CoV) is an enveloped virus that contains a single

➢The spikes of SARS-CoV consist of trimers of an S protein that is composed of two

➢Like Gardasil, this vaccine also requires three injections.

➢ This is a schematic overview of the development of a lactic acid

➢ In this procedure, it is essential that the gene encoding the target

➢ This ensures that no antibiotic resistance genes, often used as

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ Intradermal injection can achieve comparable immune responses to intramuscular

➢ As an alternative to intradermal injection, some researchers developed microneedles

➢ More recently, other scientists developed a Nanopatch technique wherein a very

➢ A single epitope may not be sufficiently immunogenic.

➢ Malaria is a parasitic disease transmitted to humans and

➢ The genus Plasmodium consists of approximately 125

➢ Because P. falciparum elicits the most severe malaria

➢ The Plasmodium life cycle is very complex. Sporozoites

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ These merosomes lodge in pulmonary capillaries and

➢ The parasite eventually forms gametocytes, that develop

➢ In the life cycle of the malaria parasite, it is the asexual

➢ In areas where malaria is endemic, some individuals show considerable resistance to

➢ It was therefore decided to chemically synthesize peptides that corresponded to small

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ Therefore, a peptide representing amino acid residues 181 to 276 of merozoite

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ Although a vaccine containing a merozoite antigen would protect against the

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ Some researchers consider using nanoparticles is as both a

➢ Therefore, researchers used a complex nanoparticle composed

➢ P. vivax is the second most serious malaria-causing parasite

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ Immunization with these nanoparticle complexes together with

➢ In addition, the immune response against this protein lasted for

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ DNA vaccination is a relatively new technique for protecting

➢ The gene in the injected DNA directs cells to produce a

➢ An advantage of genetic immunization, besides bypassing

Molecular Biotechnology: Principles and Applications Copyright © 2017 ASM Press

➢ In comparison to control mice, the nucleoprotein-injected mice were significantly

➢ One approach to deliver foreign DNA to animal cells

➢ Plasmid DNA is bound to the surfaces of these

➢ Using microparticles, it was possible to achieve the

➢ In addition the level of antibodies induced by the