Transcript Simulated HIV Lab

Virsuses:
Human Immunodeficiency Syndrome
&
Acquired Immunodeficiency Syndrome
Simulated Anti-HIV Outline
• Clinical Detection and Diagnosis of HIV and HIV exposure
– ELISA
– Western Blot
– PCR
HIV Diagnostic Tests
• ELISA
measures
Indirect evidence of
HIV exposure
HIV Diagnostic Tests
• Western Blot
• PCR
Directly measures HIV
Polymerase Chain Reaction
• Measures proviral DNA within the host DNA
Western Blot
Identifies HIV proteins
Protein ladder
Enzyme Linked Immunosorbent Assay
(ELISA)
• A diagnostic test for Antibodies to HIV
Antibodies
Antibodies are proteins produced by
our immune system that are directed
against specific antigens.
Antibodies
Antigens
Immune system
Non-self
Now apply these concepts to
the diagnostic test known as
ELISA to detect antibodies
against HIV from a biologic
fluid.
Do any of these individuals carry
antibodies to HIV?
HIV Diagnostic Tests
• ELISA
measures
Indirect evidence of
HIV exposure
HIV Diagnostic Tests
• Western Blot
• PCR
Directly measures HIV
Polymerase Chain Reaction
• Measures proviral DNA within the host DNA
Do any of these individuals carry
antibodies to HIV?
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).
1
2
OraQuick Rapid Anti-HIV Blood Test
3
20 minute test
Cost app. $15.00
Onto the ELISA lab
For Your Information and Files
Acquired Immunodeficiency Syndrome
• Normal CD4+ count
• Normal CD4+ (%)
• 500-1600/mm3
• 20-40%
• <350/mm3 begin anti-viral treatment
• AIDS
• <14% serious immune damage
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
AZT/3TC
(Combivir)
NRTI
one pill bid
(300 mg AZT/150 mg 3TC)
see above
AZT/3TC/a
bacavir
(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
lopinavir/ri
tonavir
PI
three capsules bid
(133.3 mg lopinavir/33.3 mg
ritonavir)
nausea/vomiting,
diarrhea, rash
diarrhea, nausea, weakness,
headache
Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)
Brand Name
Fuzeon™
Generic Name
Abbreviation
Experimental Code
Pharmaceutical Company
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
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
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
Squibb GSK-873,140GlaxoSmithKline 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 Tcells: BMS-488043 targets the gp120 protein, PRO-542 and TNX-355 target the CD4 protein, and SCHD,
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)
ELISA MICROTITER
PLATES
Microtiter plate
The ELISA protocol
sample
Labelled 2nd Ab
antigen
2
3
1
4
Color inducing
substrate
Results
POSITIVE ANTI-HIV
COLOR CHANGE
NEGATIVE ANTI-HIV
NO COLOR CHANGE
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).
1
2
OraQuick Rapid Anti-HIV Blood Test
3
20 minute test
Cost app. $15.00
Onto the ELISA lab
For Your Information and Files
Acquired Immunodeficiency Syndrome
• Normal CD4+ count
• Normal CD4+ (%)
• 500-1600/mm3
• 20-40%
• <350/mm3 begin anti-viral treatment
• AIDS
• <14% serious immune damage
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
AZT/3TC
(Combivir)
NRTI
one pill bid
(300 mg AZT/150 mg 3TC)
see above
AZT/3TC/a
bacavir
(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
lopinavir/ri
tonavir
PI
three capsules bid
(133.3 mg lopinavir/33.3 mg
ritonavir)
nausea/vomiting,
diarrhea, rash
diarrhea, nausea, weakness,
headache
Experimental drugs are italicized, and approved drugs are in regular, non-italicized type)
Brand Name
Fuzeon™
Generic Name
Abbreviation
Experimental Code
Pharmaceutical Company
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
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
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
Squibb GSK-873,140GlaxoSmithKline 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 Tcells: BMS-488043 targets the gp120 protein, PRO-542 and TNX-355 target the CD4 protein, and SCHD,
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)