Virsuses: Human Immunodeficiency Syndrome

1
VirsusesHuman 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?
16
(No Transcript)
17
(No Transcript)
18
FDA Approves Saliva OraQuick Rapid Test for
HIV-1, HIV-2 AntibodiesMarch 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
Acquired Immunodeficiency Syndrome

• Normal CD4 count
• Normal CD4 ()
• AIDS

• 500-1600/mm3
• 20-40
• lt350/mm3 begin anti-viral treatment
• lt14 serious immune damage

23
Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents   
Agent Type Dose Major Toxicities
AZT NRTI 300 mg bid nausea, headache,  low blood counts
ddI NRTI 125-200 mg bid or 250-400 mg qd(tablet form) diarrhea, pancreatitis,  peripheral neuropathy
ddC NRTI 0.750 mg tid diarrhea,  peripheral neuropathy
d4T NRTI 30-40 mg bid abnormal liver function tests,peripheral neuropathy
3TC NRTI 150 mg bid minor
abacavir NRTI 300 mg bid hypersensitivity reaction
tenofovir NRTI (nucleotide) 300 mg qd nausea, diarrhea, vomiting, flatulence











24
AZT/3TC (Combivir) NRTI one pill bid(300 mg AZT/150 mg 3TC) see above
AZT/3TC/abacavir(Trizivir) NRTI one pill bid(300 mg AZT/150 mg 3TC/300 mg abacavir) see above
nevirapine NNRTI 200 mg bid rash
delavirdine NNRTI 400 mg tid rash
efavirenz NNRTI 600 mg qhs rash, dizziness, impaired concentration, insomnia, abnormal dreams
saquinavir(Fortovase) PI 1200 mg tid diarrhea
ritonavir PI 600 mg bid nausea/vomiting, drug interactions
indinavir PI 800 mg tid kidney stones
nelfinavir PI 750 mg tid or 1250 mg bid diarrhea
amprenavir PI 1200 mg bid nausea/vomiting,  diarrhea, rash
lopinavir/ritonavir PI three capsules bid(133.3 mg lopinavir/33.3 mg ritonavir) diarrhea, nausea, weakness, headache
25
Experimental drugs are italicized, and approved
drugs are in regular, non-italicized type)
Brand Name Generic Name Abbreviation Experimental Code Pharmaceutical Company
Fuzeon enfuvirtide ENF T-20 Trimeris and Hoffmann-La Roche
      BMS-488043 Bristol-Myers Squibb
      GSK-873,140 GlaxoSmithKline
      PRO-542 Progenics Pharmaceuticals
      SCH-D Schering-Plough Corporation
      TNX-355 Tanox and Biogen Idec
      UK-427,857 Pfizer
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 FuzeonenfuvirtideENFT-20Tr
imeris and Hoffmann-La Roche   
BMS-488043Bristol-Myers Squibb   GSK-873,140GlaxoS
mithKline   PRO-542Progenics Pharmaceuticals   SCH
-DSchering-Plough Corporation   TNX-355Tanox and
Biogen Idec   UK-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 linkThe HIV Life Cycle (and the targets
of each class of anti-HIV drugs)
28
ELISA MICROTITER PLATES
Microtiter plate
29
(No Transcript)
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
32
(No Transcript)
33
(No Transcript)
34
FDA Approves Saliva OraQuick Rapid Test for
HIV-1, HIV-2 AntibodiesMarch 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
Acquired Immunodeficiency Syndrome

• Normal CD4 count
• Normal CD4 ()
• AIDS

• 500-1600/mm3
• 20-40
• lt350/mm3 begin anti-viral treatment
• lt14 serious immune damage

39
Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents    Table 1. For Your personal Information not for lecture. Antiretroviral Agents   
Agent Type Dose Major Toxicities
AZT NRTI 300 mg bid nausea, headache,  low blood counts
ddI NRTI 125-200 mg bid or 250-400 mg qd(tablet form) diarrhea, pancreatitis,  peripheral neuropathy
ddC NRTI 0.750 mg tid diarrhea,  peripheral neuropathy
d4T NRTI 30-40 mg bid abnormal liver function tests,peripheral neuropathy
3TC NRTI 150 mg bid minor
abacavir NRTI 300 mg bid hypersensitivity reaction
tenofovir NRTI (nucleotide) 300 mg qd nausea, diarrhea, vomiting, flatulence











40
AZT/3TC (Combivir) NRTI one pill bid(300 mg AZT/150 mg 3TC) see above
AZT/3TC/abacavir(Trizivir) NRTI one pill bid(300 mg AZT/150 mg 3TC/300 mg abacavir) see above
nevirapine NNRTI 200 mg bid rash
delavirdine NNRTI 400 mg tid rash
efavirenz NNRTI 600 mg qhs rash, dizziness, impaired concentration, insomnia, abnormal dreams
saquinavir(Fortovase) PI 1200 mg tid diarrhea
ritonavir PI 600 mg bid nausea/vomiting, drug interactions
indinavir PI 800 mg tid kidney stones
nelfinavir PI 750 mg tid or 1250 mg bid diarrhea
amprenavir PI 1200 mg bid nausea/vomiting,  diarrhea, rash
lopinavir/ritonavir PI three capsules bid(133.3 mg lopinavir/33.3 mg ritonavir) diarrhea, nausea, weakness, headache
41
Experimental drugs are italicized, and approved
drugs are in regular, non-italicized type)
Brand Name Generic Name Abbreviation Experimental Code Pharmaceutical Company
Fuzeon enfuvirtide ENF T-20 Trimeris and Hoffmann-La Roche
      BMS-488043 Bristol-Myers Squibb
      GSK-873,140 GlaxoSmithKline
      PRO-542 Progenics Pharmaceuticals
      SCH-D Schering-Plough Corporation
      TNX-355 Tanox and Biogen Idec
      UK-427,857 Pfizer
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 FuzeonenfuvirtideENFT-20Tr
imeris and Hoffmann-La Roche   
BMS-488043Bristol-Myers Squibb   GSK-873,140GlaxoS
mithKline   PRO-542Progenics Pharmaceuticals   SCH
-DSchering-Plough Corporation   TNX-355Tanox and
Biogen Idec   UK-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 linkThe HIV Life Cycle (and the targets
of each class of anti-HIV drugs)