1. Immunity Specific defenses Immunity Passive immunityActive immunity Following clinical infection Following subclinical infection Following vaccination Following administration of Immunoglobulin or antiserum Transfer of maternal Antibodies Through milk Transfer of maternal Antibodies Through placenta natural acquired

2. Viral Vaccines

3. Immunizing agents antiseraimmunuglobulinsvaccines

4. Vaccination Vaccination is a method of giving antigen to stimulate the immune response through active immunization. A vaccine is an immuno-biological substance designed to produce specific protection against a given disease. A vaccine is “antigenic” but not “pathogenic”.

5. Let’s go back in time to see how this strategy works The time: 500 B.C. The place: Greece

6. Even 2,500 Years Ago, People Knew Immunity Worked. Greek physicians noticed that people who survived smallpox never got it again. The insight: Becoming infected by certain diseases gives immunity.

7. Fast forward 2300 years pathmicro.med.sc.edu/ppt-vir/vaccine.ppt I had a brilliant idea

8. Vaccination Charles Jenner 1796 : Cowpox/Swinepox 1800’s Compulsory childhood vaccination

9. Smallpox 1% v. 25% mortality Life-long immunity UK: 1700’s China 1950 Pakistan/Afghanistan/ Ethiopia 1970 pathmicro.med.sc.edu/ppt-vir/vaccine.ppt

10. No animal reservoir Lifelong immunity Subclinical cases rare Infectivity does not precede overt symptoms One serotype pathmicro.med.sc.edu/ppt-vir/vaccine.ppt Smallpox presented many advantages that made this possible

11. As a result, after a world-wide effort Smallpox was eliminated as a human disease in 1978 pathmicro.med.sc.edu/ppt-vir/vaccine.ppt

12. Types of vaccines Live vaccines Attenuated live vaccines Inactivated (killed vaccines) Toxoids Polysaccharide and polypeptide (cellular fraction) vaccines Surface antigen (recombinant) vaccines.

13. Live vaccines Live vaccines are made from live infectious agents without any amendment. The only live vaccine is “Variola” small pox vaccine, made of live vaccinia cow-pox virus (not variola virus) which is not pathogenic but antigenic, giving cross immunity for variola.

14. Live attenuated (avirulent) vaccines Virulent pathogenic organisms are treated to become attenuated and avirulent but antigenic. They have lost their capacity to induce full-blown disease but retain their immunogenicity. Live attenuated vaccines should not be administered to persons with suppressed immune response due to: – Leukemia and lymphoma – Other malignancies – Receiving corticosteroids and anti-metabolic agents – Radiation – pregnancy

15. Live Attenuated Vaccines have several advantages Attenuated (weakened) form of the "wild" virus or bacterium Can replicate themselves so the immune response is more similar to natural infection Usually effective with one dose

16. Live Attenuated Vaccines also have several disadvantages Severe reactions possible especially in immune compromised patients Worry about recreating a wild-type pathogen that can cause disease Fragile – must be stored carefully MMWR, CDC

17. A number of the vaccines you received were live Attenuated Vaccines Viral measles, mumps, rubella, vaccinia, varicella/zoster, yellow fever, rotavirus, intranasal influenza, oral polio BacterialBCG (TB), oral typhoid

18. Inactivated (killed) vaccines Organisms are killed or inactivated by heat or chemicals but remain antigenic. They are usually safe but less effective than live attenuated vaccines. The only absolute contraindication to their administration is a severe local or general reaction to a previous dose.

19. Inactivated Vaccines Cannot replicate and thus generally not as effective as live vaccines Usually require 3-5 doses Immune response mostly antibody based Minuses

20. Inactivated Vaccines No chance of recreating live pathogen Less interference from circulating antibody than live vaccines Pluses

21. Inactivated Vaccines are also a common approach today Viral polio, hepatitis A, rabies, influenza* Bacterialpertussis*, typhoid* cholera*, plague* Whole-cell vaccines *not used in the United States

22. Other Inactivated Vaccines now contain purified proteins rather than whole bacteria/viruses Proteinshepatitis B, influenza, acellular pertussis, human papillomavirus, anthrax, Lyme Toxinsdiphtheria, tetanus

23. Sabin Polio Vaccine Attenuated by passage in foreign host (monkey kidney cells) Selection to grow in new host makes virus less suited to original host

24. Sabin Polio Vaccine Attenuated by passage in foreign host (monkey kidney cells) Selection to grow in new host makes virus less suited to original host Grows in epithelial cells Does not grow in nerves No paralysis Local gut immunity (IgA)

25. Salk Polio Vaccine Formaldehyde-fixed No reversion

26. US: Sabin attenuated vaccine ~ 10 cases vaccine-associated polio per year = 1 in 4,000,000 vaccine infections Scandinavia: Salk dead vaccine No gut immunity Cannot wipe out wt virus Polio Vaccine illustrates the pluses and minuses of live vaccines pathmicro.med.sc.edu/ppt-vir/vaccine.ppt

27. Reciprocal virus antibody titer 512 128 32 8 2 1 Serum IgG Serum IgM Nasal and duodenal IgA Nasal IgA Serum IgA Duodenal IgA Days Vaccination 4848 48969696 Killed (Salk) Vaccine Live (Sabin) Vaccine Live virus generates a more complete immune response

28. Modern molecular biology has offered new approaches to make vaccines 1.Clone gene from virus or bacteria and express this protein antigen in yeast, bacteria or mammalian cells in culture

29. Modern molecular biology has offered new approaches to make vaccines 2. Clone gene from virus or bacteria Into genome of another virus (adenovirus, canary pox, vaccinia) And use this live virus as vaccine

30. Cloned protein antigens have pluses and minuses Pluses Easily manufactured and often relatively stable Cannot “revert” to recreate pathogen Minuses Poorly immunogenic Post-translational modifications Poor CTL response

31. Viral vectors have pluses and minuses Pluses Infects human cells but some do not replicate Better presentation of antigen Generate T cell response Minuses Can cause bad reactions Can be problems with pre-exisiting immunity to virus Often can only accommodate one or two antigens

32. Toxoids They are prepared by detoxifying the exotoxins of some bacteria rendering them antigenic but not pathogenic. Adjuvant (e.g. alum precipitation) is used to increase the potency of vaccine. The antibodies produces in the body as a consequence of toxoid administration neutralize the toxic moiety produced during infection rather than act upon the organism itself. In general toxoids are highly efficacious and safe immunizing agents.

33. Polysaccharide and polypeptide (cellular fraction) vaccines They are prepared from extracted cellular fractions e.g. meningococcal vaccine from the polysaccharide antigen of the cell wall, the pneumococcal vaccine from the polysaccharide contained in the capsule of the organism, and hepatitis B polypeptide vaccine. Their efficacy and safety appear to be high.

34. Surface antigen (recombinant) vaccines. It is prepared by cloning HBsAg gene in yeast cells where it is expressed. HBsAg produced is then used for vaccine preparations. Their efficacy and safety also appear to be high.

35. Types of vaccines Live vaccines Live Attenuate d vaccines Killed Inactivated vaccines ToxoidsCellular fraction vaccines Recombinant vaccines Small pox variola vaccine BCG Typhoid oral Plague Oral polio Yellow fever Measles Mumps Rubella Intranasal Influenza Typhus Typhoid Cholera Pertussis Plague Rabies Salk polio Intra- muscular influenza Japanise encephalitis Diphtheria Tetanus Meningococcal polysaccharide vaccine Pneumococcal polysaccharide vaccine Hepatitis B polypeptide vaccine Hepatitis B vaccine

36. Routes of administration Deep subcutaneous or intramuscular route (most vaccines) Oral route (sabine vaccine, oral BCG vaccine) Intradermal route (BCG vaccine) Scarification (small pox vaccine) Intranasal route (live attenuated influenza vaccine)

37. Scheme of immunization Primary vaccination – One dose vaccines (BCG, variola, measles, mumps, rubella, yellow fever) – Multiple dose vaccines (polio, DPT, hepatitis B) Booster vaccination To maintain immunity level after it declines after some time has elapsed (DT, MMR).

38. Periods of maintained immunity due to vaccines Short period (months): cholera vaccine Two years: TAB vaccine Three to five years: DPT vaccine Five or more years: BCG vaccine Ten years: yellow fever vaccine Solid immunity: measles, mumps, and rubella vaccines.

39. Levels of effectiveness Absolutely protective(100%): yellow fever vaccine Almost absolutely protective (99%): Variola, measles, mumps, rubella vaccines, and diphtheria and tetanus toxoids. Highly protective (80-95%): polio, BCG, Hepatitis B, and pertussis vaccines. Moderately protective (40-60%) TAB, cholera vaccine, and influenza killed vaccine.

40. HIV Vaccine

41. Given that introduction, should we search for a vaccine against HIV and how would we do so?

42. This formidable array of defense mechanisms Allows HIV to avoid being suppressed by our immune system Antigenic escape Inaccessible epitopes Downregulating MHC Destruction of CD4+ T cells Integration and latency

43. An effective vaccine could have a MAJOR Impact on the future prognosis iavi.org

44. An effective vaccine must get around the strategies HIV uses to evade the immune system

45. The vaccine must be able to target conserved and essential parts of the viruses machinery Antigenic escape Inaccessible epitopes + existence of many viral strains

46. Molecular Biology of the Cell Alberts et al The vaccine must act early in the process Before the virus becomes firmly established And destroys the immune system Destruction of CD4+ T cells Integration and latency

47. There are many possible HIV Vaccine Approaches Protein subunit Synthetic peptide Naked DNA Inactivated Virus Live-attenuated Virus Live-vectored Vaccine Ramil Sapinoro, University of Rochester Medical Center

48. To begin we need to ask some key questions What should vaccine elicit?

49. To begin we need to ask some key questions What should vaccine elicit? Neutralizing antibodies to kill free virus

50. To begin we need to ask some key questions What should vaccine elicit? Neutralizing antibodies to kill free virus T cell response to kill infected cells OR

51. To begin we need to ask some key questions What should vaccine elicit? Neutralizing antibodies to kill free virus T cell response to kill infected cells OR OR BOTH?

52. The biology of HIV provides some clues

53. Long term progressors Infected with a Nef mutant virus?

54. This would generate both an antibody and a T cell response Could this be used to generate a vaccine?

55. This prompted an experiment that demonstrated the feasibility of a vaccine December 1992: Live attenuated SIV vaccine Lacking the gene Nef protected all monkeys for 2 years against massive dose of virus All controls died cell mediated immunity was key

56. However, this approach is still viewed as too risky to try on human subjects December 1992: Live attenuated SIV vaccine Lacking the gene Nef protected all monkeys for 2 years against massive dose of virus All controls died cell mediated immunity was key

57. The next efforts attempted to use recombinant viral proteins as antigens in an effort to generate neutralizing antibodies

58. VaxGen made two different forms of gp120 from different HIV strains and began human trials after chimp testing

59. Human vaccine trials are large and very expensive

60. The trial was a failure, with only minor effects seen that were viewed as statistically insignificant NY Times

61. The next approach involved using viral vectors to try to also boost the T cell response

62. Many different viral vectors are being investigated but this trial used the human cold virus called adenovirus

63. They actually used three adenoviruses carrying three different viral proteins Gag Pol Nef

64. Early results suggested the immune system was being stimulated

65. The hotly awaited results were released at the 2007 AIDS Meeting

66. You be the judge—what happened?

68. This stunning failure led to a re-thinking of the approach

69. The field has decided in part to go back to the basics: how does HIV work and how can we assess vaccine success? Questions: For a vaccine what are the measures of protection? Can we overcome polymorphism? What are the key antigens? Attenuated or killed or neither? Is Mucosal immunity critical? Should it Prevent infection or prevent disease? What are the best Animal models How does HIV kill cells anyway?

70. However trials continue, but with more focus on the details of how they affect immunity