1. Microbiology B.E Pruitt & Jane J. Stein AN INTRODUCTION TORTORA FUNKE CASE Chapter 18 Practical Applications of Immunology

2. Vaccine History Variolation: Inoculation of smallpox into skin (18 th century) Vaccination: Inoculation of cowpox into skin Herd immunity results when most of a population is immune to a disease.

3. Jenner On 14th May 1796, Edward Jenner used cowpox- infected material obtained 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 case of smallpox. He did not become infected! Edward Jenner

4. How Vaccines work Trigger your own immune response –Artificially acquired active immunity Virus, Bacterial or Toxins Attenuated - no longer virulent Inactivated or Killed - formalin, phenol or heat destroyed

5. Types of Vaccines Attenuated whole agent vaccines: –Live, attenuated (weakened) microbes - virus or bacteria –Long term immunity –May back mutate to virulent strain (rare) Inactivated (killed) vaccines: –Killed by formalin, phenol or heat –Toxoids –Not as long lasting –Safe Subunit vaccine: –Uses fragments from virus or bacteria Produced by recombinant methods Recombinvac –Safe –Clean Conjugated vaccines: –Bind to larger particle or protein to enhance antigenicity

6. 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 Principal Vaccines Used in the United States to Prevent Bacterial Diseases in Humans

7. Vaccine Schedule

8. 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) Principal Vaccines Used in the United States to Prevent Viral Diseases in Humans

9. Other Diagnostic applications: Serological Tests Diagnostic Immunological tests: Direct tests detect antigens (from patient sample) Indirect tests detect antibodies (in patient's serum)

10. Diagnostic Immunology: Precipitation Reactions Precipitation Reactions: Involve soluble antigens with antibodies Precipitin Ring test Figure 18.3

11. 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

12. Hemagglutination Hemagglutination involves agglutination of RBCs. Figure 18.7

13. Neutralization Reactions Antibodies help eliminate the harmful effect of a virus or exotoxin 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

14. Antibody Titer Is the concentration of antibodies against a particular antigen Figure 18.5

15. Complement Fixation Figure 18.9.1

16. Complement Fixation Figure 18.9.2

17. Fluorescent Antibody Techniques (Direct) Figure 18.10a

18. Fluorescent Antibody Techniques (Indirect) Figure 18.10b

19. Enzyme-Linked Immunosorbent Assay(Direct ELISA) Figure 18.12a

20. Enzyme-Linked Immunosorbent Assay (Indirect ELISA) Figure 18.12b

21. Serological Tests Figure 18.13

22. 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.