Virsuses: Human Immunodeficiency Syndrome & Acquired Immunodeficiency Syndrome

1. Virsuses:Human Immunodeficiency Syndrome& Acquired Immunodeficiency Syndrome

2. Simulated Anti-HIV Outline • Clinical Detection and Diagnosis of HIV and HIV exposure • ELISA • Western Blot • PCR

3. HIV Diagnostic Tests • ELISA measures Indirect evidence of HIV exposure

4. HIV Diagnostic Tests • Western Blot • PCR Directly measures HIV

5. Polymerase Chain Reaction • Measures proviral DNA within the host DNA

6. Western Blot Identifies HIV proteins Protein ladder

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

8. Antibodies are proteins produced by our immune system that are directed against specific antigens.

9. Antibodies Antigens Immune system Non-self

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

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

12. HIV Diagnostic Tests • ELISA measures Indirect evidence of HIV exposure

13. HIV Diagnostic Tests • Western Blot • PCR Directly measures HIV

14. Polymerase Chain Reaction • Measures proviral DNA within the host DNA

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

18. 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).

19. 1 2 OraQuick Rapid Anti-HIV Blood Test 20 minute test Cost app. $15.00 3

20. Onto the ELISA lab

21. For Your Information and Files

22. Normal CD4+ count Normal CD4+ (%) AIDS 500-1600/mm3 20-40% <350/mm3 begin anti-viral treatment <14% serious immune damage Acquired Immunodeficiency Syndrome

25. Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)

26. Interesting links on HIV • http://www.niaid.nih.gov/factsheets/aidsstat.htm • Links to global and US HIV/AIDS statistics • http://www.avert.org/pregnanc.htm • Links to HIV and pregnancy as well as numerous other links including statistics on global epidemic; HIV/AIDS quizzes and treatment. • http://www.cdc.gov/hiv/pubs/facts/transmission.htm • Links to CDC and a comprehensive fact sheet on HIV transmission

27. Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)Brand NameGeneric NameAbbreviationExperimental Code Pharmaceutical Company Fuzeon™enfuvirtideENFT-20Trimeris and Hoffmann-La Roche    BMS-488043Bristol-Myers SquibbGSK-873,140GlaxoSmithKlinePRO-542Progenics PharmaceuticalsSCH-DSchering-Plough CorporationTNX-355Tanox and Biogen IdecUK-427,857Pfizer What are Entry Inhibitors (including Fusion Inhibitors)?Entry inhibitors work by preventing HIV from entering healthy T-cells in the body. They work differently than many of the approved anti-HIV drugs – the protease inhibitors (PIs), the nucleoside reverse transcriptase inhibitors (NRTIs), and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) – which are active against HIV after it has infected a T-cell. Entry inhibitors work by attaching themselves to proteins on the surface of T-cells or proteins on the surface of HIV. In order for HIV to bind to T-cells, the proteins on HIV's outer coat must bind to the proteins on the surface of T-cells. Entry inhibitors prevent this from happening. Some entry inhibitors target the gp120 or gp41 proteins on HIV's surface. Some entry inhibitors target the CD4 protein or the CCR5 or CXCR4 receptors on a T-cell's surface. If entry inhibitors are successful in blocking these proteins, HIV is unable to bind to the surface of T-cells and gain entry into the cells. Only one entry inhibitor has been approved by the U.S. Food and Drug Administration (FDA): Fuzeon™ (T-20). This drug targets the gp41 protein on HIV's surface. Some experimental drugs target proteins on T-cells: BMS-488043 targets the gp120 protein, PRO-542 and TNX-355 target the CD4 protein, and SCH-D, GSK-873,140 and UK-427,857 target the CCR5 protein. HIV-positive people who have become resistant to PIs, NRTIs, and NNRTIs will likely benefit from the entry inhibitors because they are a different class of drugs. This is good news for HIV-positive people who have tried and failed many of the currently approved anti-HIV medications.To learn more on how HIV infects a T-cell and begins to create more viruses, and where each class of anti-HIV drugs blocks this process, click on the following lesson link:The HIV Life Cycle (and the targets of each class of anti-HIV drugs)

28. ELISA MICROTITER PLATES Microtiter plate

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

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

34. 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).

35. 1 2 OraQuick Rapid Anti-HIV Blood Test 20 minute test Cost app. $15.00 3

36. Onto the ELISA lab

37. For Your Information and Files

38. Normal CD4+ count Normal CD4+ (%) AIDS 500-1600/mm3 20-40% <350/mm3 begin anti-viral treatment <14% serious immune damage Acquired Immunodeficiency Syndrome

41. Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)

42. Interesting links on HIV • http://www.niaid.nih.gov/factsheets/aidsstat.htm • Links to global and US HIV/AIDS statistics • http://www.avert.org/pregnanc.htm • Links to HIV and pregnancy as well as numerous other links including statistics on global epidemic; HIV/AIDS quizzes and treatment. • http://www.cdc.gov/hiv/pubs/facts/transmission.htm • Links to CDC and a comprehensive fact sheet on HIV transmission

43. Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)Brand NameGeneric NameAbbreviationExperimental Code Pharmaceutical Company Fuzeon™enfuvirtideENFT-20Trimeris and Hoffmann-La Roche    BMS-488043Bristol-Myers SquibbGSK-873,140GlaxoSmithKlinePRO-542Progenics PharmaceuticalsSCH-DSchering-Plough CorporationTNX-355Tanox and Biogen IdecUK-427,857Pfizer What are Entry Inhibitors (including Fusion Inhibitors)?Entry inhibitors work by preventing HIV from entering healthy T-cells in the body. They work differently than many of the approved anti-HIV drugs – the protease inhibitors (PIs), the nucleoside reverse transcriptase inhibitors (NRTIs), and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) – which are active against HIV after it has infected a T-cell. Entry inhibitors work by attaching themselves to proteins on the surface of T-cells or proteins on the surface of HIV. In order for HIV to bind to T-cells, the proteins on HIV's outer coat must bind to the proteins on the surface of T-cells. Entry inhibitors prevent this from happening. Some entry inhibitors target the gp120 or gp41 proteins on HIV's surface. Some entry inhibitors target the CD4 protein or the CCR5 or CXCR4 receptors on a T-cell's surface. If entry inhibitors are successful in blocking these proteins, HIV is unable to bind to the surface of T-cells and gain entry into the cells. Only one entry inhibitor has been approved by the U.S. Food and Drug Administration (FDA): Fuzeon™ (T-20). This drug targets the gp41 protein on HIV's surface. Some experimental drugs target proteins on T-cells: BMS-488043 targets the gp120 protein, PRO-542 and TNX-355 target the CD4 protein, and SCH-D, GSK-873,140 and UK-427,857 target the CCR5 protein. HIV-positive people who have become resistant to PIs, NRTIs, and NNRTIs will likely benefit from the entry inhibitors because they are a different class of drugs. This is good news for HIV-positive people who have tried and failed many of the currently approved anti-HIV medications.To learn more on how HIV infects a T-cell and begins to create more viruses, and where each class of anti-HIV drugs blocks this process, click on the following lesson link:The HIV Life Cycle (and the targets of each class of anti-HIV drugs)