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VACCINES
A JOURNEY FROM JENNER TO GENES
HISTORICAL PERSPECTIVES
2
Jenner 1796 : Cowpox
• 1800’s Compulsory
childhood vaccination
• 1930’s Last natural UK
case
• 1940’s last natural US case
• 1958 WHO program
• October 1977: Last case
(Somalia)
Immunization
Immunisation is the process of conferring
increased resistance (or decreased
susceptibility) to infection
Active Vs Passive Immunization
 Active immunization
 Can be achieved through natural infection by a
microorganism or it can be acquired artificially
through the administration of vaccines
 Antibodies produced in response to an
infection e.g. natural measles virus
 Antibodies produced in response to a vaccine
(live, inactivated or toxoid)
 Individuals make their own antibodies
Active Vs Passive Immunization
Passive Immunity
 Individual gains antibodies from another
 who has produced them
 Transfer of maternal antibodies through the
placenta or in breast milk
 Administration of antibodies collected from
actively immune humans or animals e.g.
varicella zoster immunoglobulin VZIG
6
What is a Vaccine?
 A preparation of the causative agent of a
disease, specially treated for use in vaccination,
in order to induce or increase immunity
 Typically contains an agent that resembles a
disease-causing microorganism, and is often
made from Weakened or killed microorganisms
or subunits (purified protein subunits,
polysaccharides).
What is a Vaccine?
 Based on 2 key elements of adaptive
immunity:
Specificity and Memory
 Stimulation of an individual’s own
immune system to produce antibodies by
administration of a vaccine
Active Immunization through
designing Vaccines
9
Before designing any vaccine, two things
should be considered:
 Activation of humoral and cell mediated
immune response shown by a particular
antigen
 Development of immunologic memory
by the particular antigen
The Mechanism of a Vaccine
 In an ideal scenario,
whenever a vaccine is first
administered, it is
phagocytized by an antigen
presenting cell.
 Recent research suggest
that it is particularly
important that the vaccine
be taken up by a dendritic
cell.
 This is because dendritic
cells play a key role in
activating T cells, which
become helper T cells
 From there, the
activated Th cells goes
on to activate mature
B-cells.
 These activated B-cells
divides into two cell
types, antibody-
producing plasma cells
and, most importantly,
memory B cells.
The Mechanism of a
Vaccine
Importance of Secondary Immune
Response
 During the secondary immune response, the body
mounts a quicker, more robust attack on the
pathogen.
 Thus, the pathogen is cleared from the body
before it has the chance to cause an infection.
Adjuvants
 An adjuvant is a
chemical substance that
can be added to a
vaccine in order to
enhance the immune
response to the vaccine.
 There are three types of
adjuvants.
Adjuants
Aluminum Hydroxide and Aluminum Phosphate (Alum)
 Alum is an inorganic salt that
binds to proteins and causes
them to precipitate.
 Whenever the alum/vaccine
complex is injected into the
body, it slowly dissolves,
releasing the vaccine.
 Bacterial extracts can be added,
which enhances the immune
response.
 Alum is the only adjuvant
approved for use in humans.
Adjuants
Freund’s Adjuvant
 In Freund’s adjuvant, the
vaccine is suspended in oil
droplets.
 When injected into the body,
the vaccine slowly diffuses out
of the oil drop.
 Bacterial antigens can be added
in order to enhance the immune
response.
 Not used in humans because of
risk of severe inflammation.
Routes of Administration
 There are three different routes
of administration:
 Intradermal administration.
 Three types are intravenous,
intramuscular, and
subcutaneous.
 Oral administration.
 Vaccine is usually given in
liquid form.
 Foods, such as tomatoes, have
been engineered to produce a
vaccine.
 Intranasal administration.
Boosters
 For most vaccines, the
immunity against a particular
pathogen has a tendency
to wear off over time.
 In this case, a periodic
“booster” administration must
be given in order to
strengthen and lengthen
the duration of immunity.
Active immunization by
artificial means can be achieved
by:18
Attenuated organisms (avirulent)
Inactivated organisms (killed)
Purified microbial macromolecules
Cloned microbial antigens
Synthetic peptides
Anti-idiotype antibodies
Multivalent complexes
Types of Vaccines
19
 Live-attenuated
 Inactivated
 Toxoid
 Subunit
 Conjugate
 Combination
 Innovative
 Edible
20
Live-attenuated Vaccine
21
Oral Polio Vaccine (Sabin)
MMR (measles, mumps and rubella)
Varicella
BCG
Rotavirus
Oral typhoid etc.
Advantages of Attenuated Vaccines
22
•
•Raises immune response to all protective
antigens.
• More durable immunity
• Low cost
• Quick immunity in majority of vaccinees
• In case of polio and adeno vaccines, easy
administration
• Easy transport in field
•Disadvantages of Attenuated Vaccines
Mutation; reversion to virulence (often frequent)
The Milestone
23
Inactivated vaccines
 The bacteria or
virus is completely
killed using a
chemical, usually
formaldehyde
Examples
 hepatitis A
 hepatitis B
 poliovirus (Salk)
 influenza
24
25
26
Advantages of inactivated vaccines
Gives sufficient humoral immunity if boosters
given
• No mutation or reversio
Disadvantages of inactivated vaccines
• Many vaccinees do not raise immunity
• Boosters needed
• No local immunity (important)
• Higher cost
• Shortage of monkeys (polio)
• Failure in inactivation and immunization
with virulent virus
Vaccines from whole organisms
Many of the currently used vaccines to date
consist of inactivated or live but attenuated
bacterial cells or virus particles
Bacterial cells
Plague , pertusis, cholera
Inactivated
Tuberculosis
Attenuated BCG
Viral particals
Influenza , rubella virus Inactivated
Polio (Salk) Inactivated
Polio (Sabin) Attenuated
Yellow fever Attenuated
Toxoid Vaccines
 Pathogenic toxin is
modified to
harmless toxoid
 Examples of
Toxoid-based
vaccines include
tetanus and
diphtheria
28
Attenuated Vs Inactivated Vaccines
Attenuated Inactivated
Method of production:
Virulent strain grown under Virulence is inactivated
by adverse culture conditions chemical treatment or
by prolonged growth in
unnatural radiation
host or passage through
different unnatural hosts
Requirement of booster dose:
Generally single booster dose Multiple booster doses
is required are required
Stability:
Less stable More stable and
resistant
to natural temperature
Attenuated Vs Inactivated Vaccines
Type of host Immune responses:
Produce both cell mediated Mainly produces
and humoral immune response humoral response
Tendency to revert:
May revert to original virulent Does not revert to
virulent strain by recombination virulent form
with wild type strain or reverse
mutation
Attenuated Inactivated
Subunit Vaccines
• Risks associated with attenuated or
killed whole vaccines can be avoided by
subunit vaccines
• Consist of specific, purified
macromolecules derived pathogens
31
Subunit Vaccines
32
 Protein subunit (toxoid)- diphteria, tetanus
 Bacterial polysaccharide- S. pneumoniae, Nisseria
meningitidis
 Viral glycoproteins- Herpes simplex virus glycoprotein D
 Synthetic peptides produced from active epitopes of
antigen are used as vaccines.
Synthetic peptide vaccines for HIV, influenza, diphtheria
toxin, hepatitis B virus and malaria parasite are currently
being evaluated for their efficacy
Not very successful as peptides are not as immunogenic as
proteins
Subunit Vaccines
 Recombinant pathogen proteins-
Gene coding any immunogenic protein cloned by
recombinant DNA technology
E.g. hepatitis Bs Ag gene cloned in yeast
34
Recombinant Vectors Vaccines
Attenuated viruses and bacteria serve as
vectors
Genes encoding major antigens of virulent
pathogens can be introduced into these
Wide range of organisms used as vectors:
vaccinia virus,
attenuated polio virus,
canary pox virus, adenovirus,
BCG strain of Mycobacterium bovis
The Hierarchy of Vaccines
First generation vaccines are whole-organism vaccines
– either live and weakened, or killed forms.
The so-called second generation vaccines are subunit
vaccines, consisting of defined protein antigens (such
as tetanus or diphtheria toxoid)
or recombinant protein components (such as the
hepatitis B surface antigen).
DNA vaccines are third generation vaccines
DNA Vaccines - The Third Generation
Vaccines
37
• 1796 Jenner: wild type animal-adapted virus
• 1800’s Pasteur: Attenuated virus
• 1996 DNA vaccines
DNA Vaccines
Made up of plasmid that has been genetically
engineered to produce one or two specific proteins
(antigens) from a pathogen.
The vaccine DNA is injected into the cells of the
body, where the "inner machinery" of the host cells
"reads" the DNA and converts it into pathogenic
proteins.
Because these proteins are recognised as foreign,
when they are processed by the host cells and
displayed on their surface, the immune system is
alerted, which then triggers a range of immune
Applications of DNA
Vaccines
DNA vaccines are in their early phase
of clinical trials.
There are no DNA vaccines in market
at present.
At present human trials are under way
for malaria, influenza, AIDS, Ebola and
Herpes.
Advantages of DNA Vaccines
40
Plasmids are easily manufactured in large amounts
DNA is very stable
DNA resists temperature extremes so storage and
transport are straight forward. Refrigeration is
not required
Stable for storage
Elicit both humoral & cell mediated immunity
Focused on Antigen of interest
Long term immunity
Disadvantages of DNA Vaccines
Limited to protein immunogen only
Extended immunostimulation leads to
chronic inflammation
Plasmid used is resistance to antibiotics for
selection. Can raise the resistance to same
antibiotic in the host
Conjugate Vaccines
 Certain bacteria have
polysaccharide outer
coats that are poorly
immunogenic (T
independent Ags)
 By linking these outer
coats to proteins (e.g.
toxiods), the immune
system can be led to
recognize the
polysaccharide as if it
were a protein antigen
 Haemophilus influenzae
type B vaccine
42
Combination vaccines
43
 DTP - Combination vaccine prevents three diseases
in one shot
Diphtheria, Tetanus (lockjaw), Pertussis (whooping
cough)
 MMR
Measles, Mumps, Rubella
Edible vaccines
44
 Antigens or antibodies expressed in plants
 Oral administration
 Cheaper
 Exhibit good genetic stability
 Do not require special storage conditions
 Since syringes and needles are not used,
chances of infection are also less
45
Anti-idiotype vaccines
 Idiotype- unique amino acid sequence of V
domain of a given Ab – Ag binding site as well
as antigenic determinant
Generation of an effective immune
response to a dangerous pathogen
without exposing the individual to any
form of pathogen as vaccine
It is known as anti-idiotype antibodies
Vaccines of this type have been developed
against pathogens of viral, bacterial or
parasitic type
47
antibody
Anti-idiotype vaccine
epitope
Antibody with
epitope binding
site
Virus
Anti-idiotype vaccines
Anti-idiotype vaccines
48
antibody
Make antibody against
antibody idiotype
Anti-
idiotype
antibody
Anti-idiotype
antibody mimics
the epitope
Anti-idiotype vaccines
49
Anti-anti-idiotype
antibody
Anti-idiotype antibody
Use anti-idiotype antibody
as injectable vaccine
Antibody to anti-idiotype
antibody
Binds and
neutralizes virus
Anti-idiotype
antibody
Anti-anti-idiotype
antibody
Anti-anti-idiotype
antibody
Use as vaccine
VACCINES
The never ending
journey……
Why aren't vaccines available for all
diseases ?51
 The procedure for developing a vaccine is time taking and very
expensive
 Vaccines are prioritized in following order:
 Vaccines that fight diseases that cause the most deaths and
damage, like meningitis
 Vaccines that prevent severe diseases like measles and influenza
 Vaccines, like the one for rotavirus, that prevent significant
suffering
 Companies must get significant return on investment
 Some viruses mutate so quickly that traditional vaccines are
ineffective. A prime example is the HIV/AIDS virus

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Vaccine

  • 1. VACCINES A JOURNEY FROM JENNER TO GENES
  • 2. HISTORICAL PERSPECTIVES 2 Jenner 1796 : Cowpox • 1800’s Compulsory childhood vaccination • 1930’s Last natural UK case • 1940’s last natural US case • 1958 WHO program • October 1977: Last case (Somalia)
  • 3. Immunization Immunisation is the process of conferring increased resistance (or decreased susceptibility) to infection
  • 4. Active Vs Passive Immunization  Active immunization  Can be achieved through natural infection by a microorganism or it can be acquired artificially through the administration of vaccines  Antibodies produced in response to an infection e.g. natural measles virus  Antibodies produced in response to a vaccine (live, inactivated or toxoid)  Individuals make their own antibodies
  • 5. Active Vs Passive Immunization Passive Immunity  Individual gains antibodies from another  who has produced them  Transfer of maternal antibodies through the placenta or in breast milk  Administration of antibodies collected from actively immune humans or animals e.g. varicella zoster immunoglobulin VZIG
  • 6. 6
  • 7. What is a Vaccine?  A preparation of the causative agent of a disease, specially treated for use in vaccination, in order to induce or increase immunity  Typically contains an agent that resembles a disease-causing microorganism, and is often made from Weakened or killed microorganisms or subunits (purified protein subunits, polysaccharides).
  • 8. What is a Vaccine?  Based on 2 key elements of adaptive immunity: Specificity and Memory  Stimulation of an individual’s own immune system to produce antibodies by administration of a vaccine
  • 9. Active Immunization through designing Vaccines 9 Before designing any vaccine, two things should be considered:  Activation of humoral and cell mediated immune response shown by a particular antigen  Development of immunologic memory by the particular antigen
  • 10. The Mechanism of a Vaccine  In an ideal scenario, whenever a vaccine is first administered, it is phagocytized by an antigen presenting cell.  Recent research suggest that it is particularly important that the vaccine be taken up by a dendritic cell.  This is because dendritic cells play a key role in activating T cells, which become helper T cells
  • 11.  From there, the activated Th cells goes on to activate mature B-cells.  These activated B-cells divides into two cell types, antibody- producing plasma cells and, most importantly, memory B cells. The Mechanism of a Vaccine
  • 12. Importance of Secondary Immune Response  During the secondary immune response, the body mounts a quicker, more robust attack on the pathogen.  Thus, the pathogen is cleared from the body before it has the chance to cause an infection.
  • 13. Adjuvants  An adjuvant is a chemical substance that can be added to a vaccine in order to enhance the immune response to the vaccine.  There are three types of adjuvants.
  • 14. Adjuants Aluminum Hydroxide and Aluminum Phosphate (Alum)  Alum is an inorganic salt that binds to proteins and causes them to precipitate.  Whenever the alum/vaccine complex is injected into the body, it slowly dissolves, releasing the vaccine.  Bacterial extracts can be added, which enhances the immune response.  Alum is the only adjuvant approved for use in humans.
  • 15. Adjuants Freund’s Adjuvant  In Freund’s adjuvant, the vaccine is suspended in oil droplets.  When injected into the body, the vaccine slowly diffuses out of the oil drop.  Bacterial antigens can be added in order to enhance the immune response.  Not used in humans because of risk of severe inflammation.
  • 16. Routes of Administration  There are three different routes of administration:  Intradermal administration.  Three types are intravenous, intramuscular, and subcutaneous.  Oral administration.  Vaccine is usually given in liquid form.  Foods, such as tomatoes, have been engineered to produce a vaccine.  Intranasal administration.
  • 17. Boosters  For most vaccines, the immunity against a particular pathogen has a tendency to wear off over time.  In this case, a periodic “booster” administration must be given in order to strengthen and lengthen the duration of immunity.
  • 18. Active immunization by artificial means can be achieved by:18 Attenuated organisms (avirulent) Inactivated organisms (killed) Purified microbial macromolecules Cloned microbial antigens Synthetic peptides Anti-idiotype antibodies Multivalent complexes
  • 19. Types of Vaccines 19  Live-attenuated  Inactivated  Toxoid  Subunit  Conjugate  Combination  Innovative  Edible
  • 20. 20
  • 21. Live-attenuated Vaccine 21 Oral Polio Vaccine (Sabin) MMR (measles, mumps and rubella) Varicella BCG Rotavirus Oral typhoid etc.
  • 22. Advantages of Attenuated Vaccines 22 • •Raises immune response to all protective antigens. • More durable immunity • Low cost • Quick immunity in majority of vaccinees • In case of polio and adeno vaccines, easy administration • Easy transport in field •Disadvantages of Attenuated Vaccines Mutation; reversion to virulence (often frequent)
  • 24. Inactivated vaccines  The bacteria or virus is completely killed using a chemical, usually formaldehyde Examples  hepatitis A  hepatitis B  poliovirus (Salk)  influenza 24
  • 25. 25
  • 26. 26 Advantages of inactivated vaccines Gives sufficient humoral immunity if boosters given • No mutation or reversio Disadvantages of inactivated vaccines • Many vaccinees do not raise immunity • Boosters needed • No local immunity (important) • Higher cost • Shortage of monkeys (polio) • Failure in inactivation and immunization with virulent virus
  • 27. Vaccines from whole organisms Many of the currently used vaccines to date consist of inactivated or live but attenuated bacterial cells or virus particles Bacterial cells Plague , pertusis, cholera Inactivated Tuberculosis Attenuated BCG Viral particals Influenza , rubella virus Inactivated Polio (Salk) Inactivated Polio (Sabin) Attenuated Yellow fever Attenuated
  • 28. Toxoid Vaccines  Pathogenic toxin is modified to harmless toxoid  Examples of Toxoid-based vaccines include tetanus and diphtheria 28
  • 29. Attenuated Vs Inactivated Vaccines Attenuated Inactivated Method of production: Virulent strain grown under Virulence is inactivated by adverse culture conditions chemical treatment or by prolonged growth in unnatural radiation host or passage through different unnatural hosts Requirement of booster dose: Generally single booster dose Multiple booster doses is required are required Stability: Less stable More stable and resistant to natural temperature
  • 30. Attenuated Vs Inactivated Vaccines Type of host Immune responses: Produce both cell mediated Mainly produces and humoral immune response humoral response Tendency to revert: May revert to original virulent Does not revert to virulent strain by recombination virulent form with wild type strain or reverse mutation Attenuated Inactivated
  • 31. Subunit Vaccines • Risks associated with attenuated or killed whole vaccines can be avoided by subunit vaccines • Consist of specific, purified macromolecules derived pathogens 31
  • 32. Subunit Vaccines 32  Protein subunit (toxoid)- diphteria, tetanus  Bacterial polysaccharide- S. pneumoniae, Nisseria meningitidis  Viral glycoproteins- Herpes simplex virus glycoprotein D  Synthetic peptides produced from active epitopes of antigen are used as vaccines. Synthetic peptide vaccines for HIV, influenza, diphtheria toxin, hepatitis B virus and malaria parasite are currently being evaluated for their efficacy Not very successful as peptides are not as immunogenic as proteins
  • 33. Subunit Vaccines  Recombinant pathogen proteins- Gene coding any immunogenic protein cloned by recombinant DNA technology E.g. hepatitis Bs Ag gene cloned in yeast
  • 34. 34
  • 35. Recombinant Vectors Vaccines Attenuated viruses and bacteria serve as vectors Genes encoding major antigens of virulent pathogens can be introduced into these Wide range of organisms used as vectors: vaccinia virus, attenuated polio virus, canary pox virus, adenovirus, BCG strain of Mycobacterium bovis
  • 36. The Hierarchy of Vaccines First generation vaccines are whole-organism vaccines – either live and weakened, or killed forms. The so-called second generation vaccines are subunit vaccines, consisting of defined protein antigens (such as tetanus or diphtheria toxoid) or recombinant protein components (such as the hepatitis B surface antigen). DNA vaccines are third generation vaccines
  • 37. DNA Vaccines - The Third Generation Vaccines 37 • 1796 Jenner: wild type animal-adapted virus • 1800’s Pasteur: Attenuated virus • 1996 DNA vaccines
  • 38. DNA Vaccines Made up of plasmid that has been genetically engineered to produce one or two specific proteins (antigens) from a pathogen. The vaccine DNA is injected into the cells of the body, where the "inner machinery" of the host cells "reads" the DNA and converts it into pathogenic proteins. Because these proteins are recognised as foreign, when they are processed by the host cells and displayed on their surface, the immune system is alerted, which then triggers a range of immune
  • 39. Applications of DNA Vaccines DNA vaccines are in their early phase of clinical trials. There are no DNA vaccines in market at present. At present human trials are under way for malaria, influenza, AIDS, Ebola and Herpes.
  • 40. Advantages of DNA Vaccines 40 Plasmids are easily manufactured in large amounts DNA is very stable DNA resists temperature extremes so storage and transport are straight forward. Refrigeration is not required Stable for storage Elicit both humoral & cell mediated immunity Focused on Antigen of interest Long term immunity
  • 41. Disadvantages of DNA Vaccines Limited to protein immunogen only Extended immunostimulation leads to chronic inflammation Plasmid used is resistance to antibiotics for selection. Can raise the resistance to same antibiotic in the host
  • 42. Conjugate Vaccines  Certain bacteria have polysaccharide outer coats that are poorly immunogenic (T independent Ags)  By linking these outer coats to proteins (e.g. toxiods), the immune system can be led to recognize the polysaccharide as if it were a protein antigen  Haemophilus influenzae type B vaccine 42
  • 43. Combination vaccines 43  DTP - Combination vaccine prevents three diseases in one shot Diphtheria, Tetanus (lockjaw), Pertussis (whooping cough)  MMR Measles, Mumps, Rubella
  • 44. Edible vaccines 44  Antigens or antibodies expressed in plants  Oral administration  Cheaper  Exhibit good genetic stability  Do not require special storage conditions  Since syringes and needles are not used, chances of infection are also less
  • 45. 45
  • 46. Anti-idiotype vaccines  Idiotype- unique amino acid sequence of V domain of a given Ab – Ag binding site as well as antigenic determinant Generation of an effective immune response to a dangerous pathogen without exposing the individual to any form of pathogen as vaccine It is known as anti-idiotype antibodies Vaccines of this type have been developed against pathogens of viral, bacterial or parasitic type
  • 47. 47 antibody Anti-idiotype vaccine epitope Antibody with epitope binding site Virus Anti-idiotype vaccines Anti-idiotype vaccines
  • 48. 48 antibody Make antibody against antibody idiotype Anti- idiotype antibody Anti-idiotype antibody mimics the epitope Anti-idiotype vaccines
  • 49. 49 Anti-anti-idiotype antibody Anti-idiotype antibody Use anti-idiotype antibody as injectable vaccine Antibody to anti-idiotype antibody Binds and neutralizes virus Anti-idiotype antibody Anti-anti-idiotype antibody Anti-anti-idiotype antibody Use as vaccine
  • 51. Why aren't vaccines available for all diseases ?51  The procedure for developing a vaccine is time taking and very expensive  Vaccines are prioritized in following order:  Vaccines that fight diseases that cause the most deaths and damage, like meningitis  Vaccines that prevent severe diseases like measles and influenza  Vaccines, like the one for rotavirus, that prevent significant suffering  Companies must get significant return on investment  Some viruses mutate so quickly that traditional vaccines are ineffective. A prime example is the HIV/AIDS virus