Inactivated killed vaccines

							    TORTORA • FUNKE    • CASE


  Microbiology
         AN INTRODUCTION

                            B.E Pruitt & Jane J. Stein
         Chapter 18

Practical Applications of Immunology
                                     Vaccine History
                                     • Variolation: Inoculation
                                       of smallpox into skin
                                       (18th century)
                                     • Vaccination:
          QuickTime™ and a
TIFF (Uncompressed) decompress or      Inoculation of cowpox                QuickTime™ and a
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                                       into skin                  TIFF (Uncompressed) decompress or
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                                     • Herd immunity results
                                       when most of a
                                       population is immune to
                                       a disease.
• On 14th May 1796, Edward
                                 Jenner
  Jenner used cowpox-                                      QuickTime™ and a
  infected material obtained                     TIFF (Un compressed) decompre ssor
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  from the hand of Sarah
  Nemes, a milkmaid from his
  home village of Berkley in
  Gloucestershire to
  successfully vaccinate 8
  year old James Phipps. On
  1st July 1796, Jenner
  challenged the boy by
  deliberately inoculating him
  with material from a real                        QuickTime™ an d a

  case of smallpox.He did not             TIFF (Uncompressed) decompressor
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  become infected!
                    How
• Trigger your own immune                   Vaccines work
  response
   – Artificially acquired active
     immunity

                                                                 QuickTime™ and a
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                QuickTime™ and a
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         are neede d to see this picture.    Virus, Bacterial or Toxins
                                             •Attenuated - no longer virulent
                                             •Inactivated or Killed - formalin, phenol or heat
                                             destroyed
•   Attenuated whole agent vaccines:
     –                                 Types of Vaccines
         Live, attenuated (weakened) microbes - virus or bacteria
     –   Long term immunity
     –   May back mutate to virulent strain (rare)                                                               Qui ckTi me™ and a
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•   Inactivated (killed) vaccines:                                                                       are ne ede d to see thi s pi cture.


     –   Killed by formalin, phenol or heat
     –   Toxoids
     –   Not as long lasting
     –   Safe
•   Subunit vaccine:                                               Qui ckTi me™ and a
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     –   Uses fragments from virus or bacteria             are ne ede d to see thi s pi cture.

           •   Produced by recombinant methods Recombinvac
     –   Safe
     –   Clean
•   Conjugated vaccines:
     –   Bind to larger particle or protein to enhance antigenicity
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  Principal Vaccines Used in the United
 States to Prevent Bacterial Diseases in
                        Humans
• DTaP - Trivalent (three in one)
   – Diphtheria: Purified diphtheria toxoid
   – Pertussis: Acellular fragments of B. pertussis
   – Tetanus: Purified tetanus toxoid
• Meningococcal meningitis: Purified polysaccharide from N.
  meningitidis
• Haemophilus influenzae type b meningitis: Polysaccharides
  conjugated with protein
• Pneumococcal conjugate vaccine: S. pneumoniae antigens
  conjugated with protein
Vaccine Schedule
    Principal Vaccines Used in the United
     States to Prevent Viral Diseases in
•                         Humans
    Smallpox: Live vaccinia virus
•   Poliomyelitis: Inactivated virus
•   Rabies: Inactivated virus
•   Hepatitis A: Inactivated virus
•   Influenza: Inactivated or attenuated virus
•   Measles: Attenuated virus
•   Mumps: Attenuated virus
•   Rubella: Attenuated virus
•   Chickenpox: Attenuated virus
•   Hepatitis B: Antigenic fragments (recombinant vaccine)
        Other Diagnostic
Diagnostic Immunological tests: applications:
                   Serological patient sample)
• Direct tests detect antigens (from Tests
• Indirect tests detect antibodies (in patient's serum)
    Diagnostic Immunology: Precipitation
                 Reactions
Precipitation Reactions:
• Involve soluble
  antigens with
  antibodies
• Precipitin Ring test




                                      Figure 18.3
             Agglutination Reactions
• Involve particulate
  antigens and
  antibodies
• Antigens may be:
• On a cell (direct
  agglutination)
• Attached to latex
  spheres (indirect or
  passive
  agglutination)


                                       Figure 18.4
                 Hemagglutination
• Hemagglutination involves agglutination of RBCs.




                                                     Figure 18.7
• Antibodies help eliminate the harmful effect of a virus or
  exotoxin
             Neutralization Reactions
• Viral hemagglutination inhibition tests for the presence
  of antibodies in a patients serum by the antibodies' ability
  to prevent viruses from agglutinating RBCs.




                                                          Figure 18.8b
                 Antibody Titer


• Is the
  concentration of
  antibodies against
  a particular antigen




                                  Figure 18.5
Complement Fixation




                      Figure 18.9.1
Complement Fixation




                      Figure 18.9.2
Fluorescent Antibody Techniques (Direct)




                                    Figure 18.10a
Fluorescent Antibody Techniques
            (Indirect)




                                  Figure 18.10b
Enzyme-Linked Immunosorbent
    Assay(Direct ELISA)




                              Figure 18.12a
Enzyme-Linked Immunosorbent Assay
          (Indirect ELISA)




                              Figure 18.12b
Serological Tests




                    Figure 18.13
         Serological Tests: Summary
• Precipitation: Soluble antigens
• Agglutination: Particulate antigens
• Hemagglutination: Agglutination of RBCs
• Neutralization (inhibition): Inactivates toxin or virus
• Fluorescent-antibody technique: Antibodies linked to
  fluorescent dye
• Complement fixation: RBCs are indicator
• ELISA: Enzyme linked to antibody amplifies results for
  easier visibility and more sensitivity.