1. Practical Applications of Immunology
Chapter 18
Practical Applications of Immunology

2. Q&A
The streptococci shown in the microscope here would be visible even if fluorescent antibodies were not attached to them. Why would this test be even more valuable for testing for such organisms as the rabies virus?

3. Vaccines 18-1 Define vaccine. 18-2 Explain why vaccination works.
18-3 Differentiate the following, and provide an example of each: attenuated, inactivated, toxoid, subunit, and conjugated vaccines.
18-4 Contrast subunit vaccines and nucleic acid vaccines.

4. Vaccines
18-5 Compare and contrast the production of whole-agent vaccines, recombinant vaccines, and DNA vaccines.
18-6 Define adjuvant.
18-7 Explain the value of vaccines, and discuss acceptable risks for vaccines.

5. History of Vaccines
Variolation: Inoculation of smallpox into skin (18th century)
Vaccination:
Inoculation of cowpox virus into skin (Jenner)
Inoculation with rabies virus (Pasteur)

6. Measles in the United States, 1960–2007
Clinical Focus, p. 505

7. Measles Worldwide
Kills 600 children per day
Clinical Focus, p. 505

8. Vaccines Used to Prevent Bacterial Diseases
Diphtheria
Purified diphtheria toxoid
Meningococcal meningitis
Purified polysaccharide from Neisseria meningitidis
Pertussis (whooping cough)
Killed whole or acellular fragments of Bordetella pertussis
Pneumococcal pneumonia
Purified polysaccharide from 7 strains of Streptococcus pneumoniae
Tetanus
Purified tetanus toxoid
Haemophilus influenzae type b meningitis
Polysaccharide from Haemophilus influenzae type b conjugated with protein to enhance effectiveness

9. Vaccines Used to Prevent Bacterial Diseases
Influenza
Injected vaccine, inactivated virus (nasally administered: attenuated virus)
Measles
Attenuated virus
Mumps
Rubella
Chickenpox
Poliomyelitis
Killed virus

10. Vaccines Used to Prevent Bacterial Diseases
Rabies
Killed virus
Hepatitis B
Antigenic fragments of virus
Hepatitis A
Inactivated virus
Smallpox
Live vaccinia virus
Herpes zoster
Attenuated virus
Human papillomavirus

11. Vaccines for Persons Aged 0–6 Years
Hepatitis B
Rotavirus
DTP
Haemophilus influenzae b
Pneumococcal
Inactivated poliovirus
Influenza
MMR
Varicella
Hepatitis A
Meningococcal
ANIMATION Vaccines: Function

12. What is the etymology (origin) of the word vaccine? 18-1
Vaccination is often the only feasible way to control most viral diseases; why is this? 18-2

13. Types of Vaccines Attenuated whole-agent vaccines
MMR
Inactivated whole-agent vaccines
Salk polio
Toxoids
Tetanus

14. Types of Vaccines Subunit vaccines Nucleic acid (DNA) vaccines
Acellular pertussis
Recombinant hepatitis B
Nucleic acid (DNA) vaccines
West Nile (for horses)
ANIMATION Vaccines: Types

15. Vaccine Development
Culture pathogen
Figure 18.1

16. Culturing Viruses
Figure 13.7

17. The Development of New Vaccines
Culture pathogen
rDNA techniques
In plants
Adjuvants
Deliver in combination

18. Safety of Vaccines
Therapeutic index =
Risk vs. benefit

19. Experience has shown that attenuated vaccines tend to be more effective than inactivated vaccines. Why? 18-3
Which is more likely to be useful in preventing a disease caused by an encapsulated bacterium such as the pneumococcus: a subunit vaccine or a nucleic acid vaccine? 18-4

20. Which type of vaccine did Louis Pasteur develop, whole-agent, recombinant, or DNA? 18-5
What is the derivation of the word adjuvant?
18-6
What is the name of a currently used oral vaccine that occasionally causes the disease it is intended to prevent? 18-7

21. Diagnostic Immunology
18-8 Differentiate sensitivity from specificity in a diagnostic test.
18-9 Define monoclonal antibodies, and identify their advantage over conventional antibody production.
18-10 Explain how precipitation reactions and immunodiffusion tests work.
18-11 Differentiate direct from indirect agglutination tests.
18-12 Differentiate agglutination from precipitation tests.
18-13 Define hemagglutination.
18-14 Explain how a neutralization test works.

22. Diagnostic Immunology
18-15 Differentiate precipitation from neutralization tests.
18-16 Explain the basis for the complement-fixation test.
18-17 Compare and contrast direct and indirect fluorescent-tests.
18-18 Explain how direct and indirect ELISA tests work.
18-19 Explain how Western blotting works.
18-20 Explain the importance of monoclonal antibodies.

23. Diagnostic Immunology
Sensitivity: Probability that the test is reactive if the specimen is a true positive
Specificity: Probability that a positive test will not be reactive if a specimen is a true negative
Koch’s experiment
Guinea pigs with TB injected with Mycobacterium tuberculosis: Site became red and slightly swollen

24. Monoclonal Antibodies (Mabs)
Hybridoma: “Immortal” cancerous B cell fused with an antibody-producing normal B cell, produces
Monoclonal antibodies

25. Monoclonal Antibodies
Figure 18.2

26. Monoclonal Antibodies
Figure 18.2

27. Monoclonal Antibodies
Figure 18.2

28. Monoclonal Antibodies (Mabs)
Muromonab-CD3: For kidney transplant
Infliximab: For Crohn’s disease
Ibritumomab + rituximab: For non-Hodgkin’s lymphoma
Trastuzumab: Herceptin for breast cancer

29. Monoclonal Antibodies
Chimeric Mabs: Genetically modified mice that produce Ab with a human constant region
Humanized Mabs: Mabs that are mostly human, except for mouse antigen–binding
Fully human antibodies: Mabs produced from a human gene on a mouse

30. What property of the immune system suggested its use as an aid for diagnosing disease: specificity or sensitivity? 18-8
The blood of an infected cow would have a considerable amount of antibodies against the infectious pathogen in its blood. How would an equivalent amount of monoclonal antibodies be more useful? 18-9

31. Precipitation Reactions
Reaction of soluble antigens with antibodies
Figure 18.4

32. A Precipitation Curve
Figure 18.3

33. Immunoelectrophoresis
Figure 10.12

34. Agglutination Reactions
Particulate antigens and antibodies
Antigens on a cell (direct agglutination)
Figure 18.5

35. Indirect Agglutination
Figure 18.7

36. Antibody Titer
Concentration of antibodies against a particular antigen
Figure 18.6

37. Hemagglutination Hemagglutination involves agglutination of RBCs
Some viruses agglutinate RBCs in vitro
Figure 18.8

38. Viral Hemagglutination-Inhibition
Hemagglutination involves agglutination of RBCs
Some viruses agglutinate RBCs in vitro
Antibodies prevent hemagglutination
Figure 18.9b

39. Neutralization Reactions
Eliminate the harmful effect of a virus or exotoxin
Figure 18.9a

40. Why does the reaction of a precipitation test become visible only in a narrow range? 18-10
Why wouldn’t a direct agglutination test work very well with viruses? 18-11
Which test detects soluble antigens, agglutination or precipitation? 18-12
Certain diagnostic tests require red blood cells that clump visibly. What are these tests called? 18-13
In what way is there a connection between hemagglutination and certain viruses? 18-14
Which of these tests is an antigen–antibody reaction: precipitation or viral hemaglutination inhibition? 18-15

41. Complement Fixation Test
Figure 18.10

42. Complement Fixation Test
Figure 18.10

43. Fluorescent-Antibody (FA) Techniques
Direct FA
Figure 18.11a

44. Indirect Fluorescent-Antibody Test
Figure 18.11b

45. Q&A
The streptococci shown in the microscope here would be visible even if fluorescent antibodies were not attached to them. Why would this test be even more valuable for testing for such organisms as the rabies virus?

46. Fluorescence-Activated Cell Sorter (FACS)
Figure 18.12

47. Enzyme-Linked Immunosorbent Assay
Also called ELISA
Enzyme linked to Ab is the indicator

48. Direct ELISA
Figure 18.14a

49. Indirect ELISA
Figure 18.14b

50. Serological Tests
Figure 18.13

51. Serological Tests Direct tests detect antigens (from patient sample)
Indirect tests detect antibodies (in patient’s serum)

52. Serological Tests Precipitation: Soluble antigens
Agglutination: Particulate antigens
Hemagglutination: Agglutination of RBCs
Neutralization: Inactivates toxin or virus
Fluorescent-antibody technique: Antibodies linked to fluorescent dye
Complement fixation: RBCs are indicator
ELISA: Peroxidase enzyme is the indicator

53. Why is complement given its name? 18-16
Which test is used to detect antibodies against a pathogen: the direct or the indirect fluorescent-antibody test? 18-17
Which test is used to detect antibodies against a pathogen, direct or the indirect ELISA test? 18-18
How are antibodies detected in Western blotting? 18-19
How has the development of monoclonal antibodies revolutionized diagnostic immunology? 18-20

54. Question 1
Patient’s serum, influenza virus, sheep RBCs, and anti-sheep RBCs are mixed in a tube
Influenza virus agglutinates RBCs
What happens if the patient has antibodies against influenza virus?

55. Question 2
Patient’s serum, Chlamydia, guinea pig complement, sheep RBCs, and anti-sheep RBCs are mixed in a tube
What happens if the patient has antibodies against Chlamydia?