1. Virsuses: Human Immunodeficiency Syndrome & Acquired Immunodeficiency Syndrome

2. Outline Introductory Comments –Virus versus bacteria –Examples of DNA and RNA containing viruses –How RNA containing viruses can replicate Origins of HIV –Dr. Hahn and colleagues Characteristics of HIV Particle –Glycoproteins –Viral genes Laboratory Diagnostic Tests –PCR –Western Blot –ELISA -------------------------------------------------- Part 2 ------------------------- Treatment Transmission Prevention

3. Bacteria versus Viruses

4. Note: These STDs are bacterial infections! SYPHILIS –Bacterium: Treponema pallidum GONORRHEA –Bacterium: Neisseria gonorrhoeae

5. Viruses DNA containing RNA containing

6. DNA containing viruses Herpes virus Small pox virus Hepatitis B adenovirus

7. RNA containing viruses Hepatitis A and C Rabies Ebola Measles Mumps Polio HIV Rabies virus Measles on the back

8. HIV

9. Examples of how RNA viruses replicate Negative strand Positive strand Retro

11. RETROVIRAL REPLICATION

12. HUMAN IMMUNODEFICIECY VIRUS Family: retrovirus Genus: lentivirus Species: HIV 1 and HIV 2 Groups and Subtypes: variety

13. “Predominant Subtypes of Group M” Africa Americas China (southeast) Europe India Japan Thailand A;D;C B C B C B E

14. Dr. Beatrice Hahn & colleagues: Drs. Li and Keele

15. Greater spot nose monkey Red Cap Mangabey monkey SIV cpz almost identical to HIV 1 Humans

16. Pan troglodytes troglodytes Simian Virus (SIV cpz) almost identical to HIV 1

17. Sooty mangabey monkey Simian Virus corresponds to HIV2

18. HIV: Structure and mechanism of replication

19. HIV-1 PARTICLE

20. HIV Glycoproteins Lipid Bilayer contains glycoproteins –Gp 160 –Gp 120 –Gp 41

21. T lymphocyte in green and HIV in red

22. HIV and helper T cells HIV-1 HOST CELLS : helper T lymphocyte, macrophages http://highered.mcgraw- hill.com/sites/0072495855/student_view0/chapter24/animat ion__hiv_replication.html

23. Budding of HIV from T lymphocyte HIV

24. Mechanism of Infection VIRAL RNA PROVIRAL DNA Infects T h cell Integrates into host DNA “activated” LYTIC STAGE storage LYSOGENIC STAGE

25. Co-Receptors CCR5 or CXCR4 (fusin) Binds naturally occurring chemokines

26. LYTIC STAGE

28. The key HIV genes

29. HIV Diagnostic Tests Western Blot PCR Directly measures HIV

30. Polymerase Chain Reaction Measures proviral DNA within the host DNA

31. Western Blot Identifies HIV proteins Protein ladder

32. HIV Diagnostic Tests ELISA Indirect evidence of HIV exposure measures

33. A diagnostic test for Antibodies to HIV Enzyme Linked Immunosorbent Assay (ELISA) Antibodies

34. *Antibodies are proteins produced by our immune system that are directed against specific antigens. (Antibody interactions with antigens will also be discussed in our “immungenetics lecture”)

35. Antibodies Antigens Immune system Non-self

36. Now apply these concepts to the diagnostic test known as ELISA to detect antibodies against HIV from a biologic fluid.

37. Do any of these individuals carry antibodies to HIV?

38. ELISA MICROTITER PLATES Microtiter plate

40. The ELISA protocol 1 2 3 4 antigen sample Labelled 2 nd Ab Color inducing substrate

41. Results POSITIVE ANTI-HIV COLOR CHANGE NEGATIVE ANTI-HIV NO COLOR CHANGE

44. FDA Approves Saliva OraQuick Rapid Test for HIV-1, HIV-2 Antibodies [March 29, 2004] (similar test is also available for blood samples, see next slides).

45. OraQuick Rapid Anti-HIV Blood Test 12 3 20 minute test Cost app. $15.00

46. Outline Introductory Comments –Virus versus bacteria –Examples of DNA and RNA containing viruses –How RNA containing viruses can replicate Origins of HIV –Dr. Hahn and colleagues Characteristics of HIV Particle –Glycoproteins –Viral genes Laboratory Diagnostic Tests –PCR –Western Blot –ELISA -------------------------------------------------- Part 2 ------------------------- Treatment Transmission Prevention