Retroviridae

Lentivirus(genus)
Human Immunodeficiency Virus (species)
People living with HIV in 2015 36.7 million world-wide

New HIV infections in 2015 2.1 million world-wide

Deaths due to AIDS in 2015 1.1 million

Both HIV-1 and HIV-2 originated in non-
human primates in west-central Africa
ORIGIN OF HIV
HIV 2 is still predominantly in Africa
In USA, AIDS was first clinically observed in 1981

• The initial cases were a cluster of injection drug users and gay
men with no known cause of impaired immunity who showed
symptoms of Pneumocystis carinii pneumonia (PCP), a rare
opportunistic fungal infection that was known to occur in people
with very compromised immune systems.
• Soon thereafter, additional gay men developed a previously rare
skin cancer called Kaposi's sarcoma (KS).
• Many more cases of PCP and KS emerged, alerting U.S. Centers
for Disease Control and Prevention (CDC) and a CDC task force
was formed to monitor the outbreak

HIV-1-associated wasting
(Cachexia) + Kaposi’s Sarcoma
 HIV historic perspective
1983/84: Isolation and Propagation of a Novel Human Retrovirus
Human Immunodeficiency Virus
(1983-Gallo, 1984-Barre-Sinoussi)

Clear Association between HIV-1 Infection, Immunodeficiency and Wasting

Disease and Acquired Immunodeficiency Syndrome (AIDS)

1984: CD4 recognized as HIV-1 receptor on T cells

1985: ELISA for routine antibody screening of blood supply

1987: First antiviral drug, AZT, inhibits reverse transcriptase

1994: Highly active antiretroviral treatment (HAART)

Objectives

• Describe the structure of HIV

• Describe the replication cycle of HIV
• Describe the stages of HIV infection
• Describe the disease mechanism of HIV
• Briefly list the steps targeted by HIV drugs
 Unique
features of
Retroviridae

• Enveloped with a capsid containing two copies of positive-strand

RNA genome
• RTase and integrase enzymes are carried in the virion
• Replication proceeds through a DNA intermediate, termed a
provirus
• The provirus integrates randomly into the host chromosome and
becomes a cellular gene
• Tx of the genome is regulated by the interaction of the host Tx
factors with promoter and enhancer elements in the LTR portion of
the genome
• HIV is easily inactivated and must be transmitted in bodily fluids.
 Simple retroviruses encode gag, pol, and env genes.

5’LTR 3’LTR

gag
pol
env

Enzymes

Structural Proteins Envelope Proteins

Complex retroviruses also encode accessory genes.

HIV is a complex virus

Env proteins:
gp120, gp41
Enzymes:
Core, structural protease,
proteins Rev- transport mRNA to cytoplasm
integrase,
RTase Nef-
TAT- upregulate Tx
downregulate
MHC I, CD4
 HIV replication cycle

• Entry of HIV into cells

• Reverse transcription
• Integration
• Transcription/Replication
• Translation
• Assembly and budding
Entry of HIV into cells

1) gp120 binds to CD4

(on T cell; Mo/MØ)

2) gp120 then change its

shape and binds to
chemokine
receptor (CCR5 or
CXCR4)

3) gp41 becomes exposed

and induces viral fusion
at normal body pH
 Reverse transcription
and integration

• RTase makes DNA strand

complementary to genomic RNA

• DdDpolymerase and RNAse

activities allow formation of double-
stranded viral cDNA

• Viral cDNAis integrated into host

chromosome

http://www.youtube.com/watch?v=RO8MP3wMvqg
Transcription/Replication/
Translation of HIV genome
• Usually in activated T
cells
• T cells are activated by
HIV or by being primed
for infection

Viral mRNA and genomic RNA

is made and transported to the
cytoplasm

Late proteins, Gag, Env and Pol are

translated as polyproteins
Assembly and Budding

Env polyprotein(gp160) is cleaved into

gp120 and gp41 in the cell

During budding and release:

Gag polyprotein is cleaved into

matrix and nucleocapsid

Pol polyprotein is cleaved into

protease, reverse transcriptase and
integrase

Virus can cause syncytia formation

with subsequent lysis of these cells.
 mRNA

protein

* AUG start cod

HIV infection 2. Massive viremia leads to
disseminated infection of
other lymphoid organs as well
1. Mucosal DC as macrophages (microglial
deliver HIV into cells) in the brain.
lymph nodes
where it can 4. CD4 T cells most
infect CD4 T likely to be killed by
cells. Mucosal HIV are those
LN are currently fighting
especially HIV or other
affected. infection (because
they are activated
and support
replication of HIV).
3. Infection of
cells of the brain
including
microglial cell and
neurons causes
the release of 5. Diseases
neurotoxic related to AIDS
substances and mainly consist of
promote opportunistic
inflammatory infections,
response in the cancers and CNS
brain. pathology.
Disease progression:

Stage 1/Acute phase

• Initial HIV symptoms occur 2-4 weeks after
infection, resemble influenza or mono

• Symptoms subside after several weeks

Stage 2/Chronic phase

• Asymptomatic infection characterizes by
lymphadenopathy that can last for years.
• CD4+Th cells, CD8+CTL, and antibodies
control viral load but do not clear the virus.
• At this time virus continues to replicate in the
lymph nodes.
• When CD4 T cell count drops below 450 cells
per microliter, opportunistic infection may Asymptomatic phase
begin to appear, and increased number of viral Flu-like
disease
RNA appears in the blood. 50-80% Symptomatic phase
of cases
Stage 3/AIDS AIDS
• When T cell count drops below 200/mL then
more serious infections occur such as
pneumonia, PML, CMV, wasting syndrome,
Kaposi’s sarcoma.
Immune response to HIV:
Opportunistic infections
Immune response unable to clear HIV
or limit disease progression to AIDS
1. Subversion
• CD4 T cells are killed directly by HIV
• Indirect killing  Inflammation, Apoptosis
• Homeostatic deregulation (damage of lymphoid
organs ex: thymus, lymph node)

2. Mutations during replication of RNA genome can lead to

escape of HIV from the immune response epitope recognition.

3. Latency - There is latent viral reservoir in lymphoid tissue

4. Inhibitory molecules - long term exposure to virus causes

immune cells to upregulate inhibitory receptors
HIV budding from T cells
 Apoptosis

Uninfected AIDS

CD4+
T cell
 How does HIV Kill CD4+ T cells?
Fibrosis

Uninfected AIDS

Collagen
Collagen
Collagen
Type
TypeIII
Type
Immune response unable to clear HIV
or limit disease progression to AIDS
1. Subversion
• CD4 T cells are killed directly by HIV
• Indirect killing  Inflammation, Apoptosis
• Homeostatic deregulation (damage of lymphoid
organs ex: thymus, lymph node)

2. Mutations during replication of RNA genome can lead to

escape of HIV from the immune response epitope recognition.

3. Latency - There is latent viral reservoir in lymphoid tissue

4. Inhibitory molecules - long term exposure to virus causes

immune cells to upregulate inhibitory receptors
Mutations
Immune response unable to clear HIV
or limit disease progression to AIDS
1. Subversion
• CD4 T cells are killed directly by HIV
• Indirect killing  Inflammation, Apoptosis
• Homeostatic deregulation (damage of lymphoid
organs ex: thymus, lymph node)

2. Mutations during replication of RNA genome can lead to

escape of HIV from the immune response epitope recognition.

3. Latency - There is latent viral reservoir in lymphoid tissue

4. Inhibitory molecules - long term exposure to virus causes

immune cells to upregulate inhibitory receptors
Latency
Immune response unable to clear HIV
or limit disease progression to AIDS
1. Subversion
• CD4 T cells are killed directly by HIV
• Indirect killing  Inflammation, Apoptosis
• Homeostatic deregulation (damage of lymphoid
organs ex: thymus, lymph node)

2. Mutations during replication of RNA genome can lead to

escape of HIV from the immune response epitope recognition.

3. Latency - There is latent viral reservoir in lymphoid tissue

4. Inhibitory molecules - long term exposure to virus causes

immune cells to upregulate inhibitory receptors
Inhibitory PD-1
Receptors
2B4
CTLA-4

ILT4 KLRG1

Immune
TIM3 Cell CLEC12A

LAG3
Means of HIV escape from the immune response
Characteristic Function
Infection of lymphocytes and Inactivation of key elements of immune
macrophages defense

Inactivation of CD4 helper cells Loss of activators of immune system

Antigenic drift of gp120 Evasion of Ab detection

Heavy glycosylation of gp120 Evasion of Ab detection

Infection of naïve, resting memory T Virus remains latent, avoid immune

cells detection

Long term exposure to virus causes Immune defenses are prematurely

immune cells to upregulate turned off
inhibitory receptors
The destruction of immune function as a result of HIV
infection leads to increased susceptibility to opportunistic
infection and eventually to death
Passage of the virus from mother to fetus

• In the absence of treatment, the risk

of HIV transmission from mother to
child is 25-30%.
•
Antiretrovirals medication lower
mother-to-infant transmission to less
than 2%.
•
The vast majority of transmission of
HIV from mother to child occurs at
the time of birth when the
membrane is ruptured and the blood
systems can mix.
 HIV Detection Assays
1) anti-HIV-1 antibodies (source: serum) (method: ELISA)

1) Anti-HIV-1 antibodies (source: serum) (method: Western)

2) Viral RNA (source: blood) (method: PCR)

3) Viral DNA (source: cells) (method: PCR)

4) p24 (source: blood) (method: ELISA)

2014 New CDC HIV Testing Recommendations
Anti-HIV Therapy
 HIV treatment
• Nucleoside RTase inhibitors (NRTIs)

• Non-nucleoside RTase inhibitor (NNRTIs)

- helped stop perinatal transmission

• Protease inhibitor (PIs)

• Integrase inhibitor (IIs)

• CCR5 antagonists

• Fusion inhibitor target gp41

These drugs only work in active cells

Anti-HIV drugs inhibit steps of virus life cycle
Fusion/Binding (2003)

X
Nucleoside (1987)
X
Non-nucleoside (1996)

X
Integrase (2007)
X
Protease (1995)

http://Open2.net
Current ARV Medications
PI
• Atazanavir(ATV)
NRTI • Darunavir(DRV)
• Abacavir (ABC) • Fosamprenavir(FPV)
• Didanosine (ddI) • Indinavir(IDV)
• Emtricitabine (FTC) • Lopinavir(LPV)
• Lamivudine (3TC) • Nelfinavir(NFV)
• Stavudine (d4T) • Ritonavir(RTV)
• Tenofovir (TDF) • Saquinavir(SQV)
• Zidovudine (AZT, ZDV) • Tipranavir(TPV)

NNRTI Integrase Inhibitor (II)

• Delavirdine (DLV) • Raltegravir(RAL)
• Efavirenz (EFV)
• Etravirine (ETR) Fusion Inhibitor
• Nevirapine (NVP) • Enfuvirtide(ENF, T-20)
• Rilpivirine (RPV)
CCR5 Antagonist
• Maraviroc(MVC)

HAART therapy = 2 NRTIs as a "backbone" along with 1 NNRTI, PI or INSTI

Reduction in viral load with HAART
Promising HIV ‘Vaccine’ Inactivates All Virus Strains, Prevents Infection
BY DOUGLAS MAIN 2/18/15 AT 8:13 AM
Virus

Normal viral
attachment

Cell

The HIV is very variable and mutates all the time, but it has two pockets that are pretty invariable and bind
CD4 (blue) and CCR5 (green). When CD4 is bound, the CCR5-binding pocket becomes accessible to bind
CCR5

Viral
attachment
is blocked
by eCD4-Ig

A piece of CD4 (blue) is attached to the synthetic antibody, binding to the CD4 circle exposes the second
pocket on gp120 but the Ab also has the CCR5 piece (green oval) attached to it, so that piece then plugs
the second pocket on HIV. Since eCD4-Ig attacks both pockets, it’s unlikely to have both mutate during
replication. This protein will be given intramuscular not IV. It will be given as part of a genetic construct
inside the AAV virus. The muscle cells will use the DNA encoding eCD4-Ig and synthesize the protein.
 Promising HIV ‘Vaccine’ Inactivates All Virus Strains, Prevents Infection
BY DOUGLAS MAIN 2/18/15 AT 8:13 AM

| Vaccination versus gene therapy. a, HIV infection begins with the virus’s envelope protein binding to CD4 and CCR5 molecules
on the surface of T cells. Most current strategies aimed at conferring protection against HIV focus on vaccines that are designed to
prevent this binding by producing antibodies that bind to structures shared by the envelope of many HIV strains. However,
antibodies bind only to small sections of the envelope, and the virus can evolve to shield these regions from antibody binding. b,
An alternative approach proposed to stop virus binding to host cells is to use artificial constructs of human CD4 attached to
immunoglobulin molecules. These CD4–Ig constructs will bind many viral strains, but they may expose the CCR5 binding site on
the envelope protein and thus actually facilitate binding of the virus to CCR5 on the host cell. c, Gardner et al.2 present an
alternative construct, eCD4–Ig, which contains both CD4 and a mimetic of CCR5 and therefore blocks both points of viral binding.
http://www.youtube.com/watch?v=RO8MP
3wMvqg