Human Immunodeficiency Virus

(HIV)

DR. ELENEUS ELIAS

DEPARTMENT OF BIOCHEMISTRY
 CONTENT
 Introduction
 Epidemiology
 History and discovery of HIV
 Etiology of HIV/AIDS
 Types of HIV viruses
 Structure of HIV
 Virus life cycle
 Transmission of HIV
 Natural History of HIV infection
 Diagnosis of HIV infections
 Treatment and ART
 HIV prevention
 Summary
 Introduction
• AIDS is a retroviral disease caused by human
immunodeficiency virus (HIV).
• The disease is characterized by immunosuppression,
secondary neoplasma and neurological
manifestations.
• Despite the absence of a cure, the natural history of the
disease has radically and now changed.
• Currently persons with HIV infection who are treated
before significant immunosuppression can expect a life
expectancy approaching that of the general
population.
 Epidemiology
By the end of 2021, the reported statistics on the global burden of
HIV were the following;
 38.4 million adults and 1.7 million children (<15 years old)
were living with HIV/AIDS.
 1.5 million adults and 160,000 children had been newly
infected with HIV that year.
 552,000 adults and 98,000 children died of AIDS that year
 In Tanzania by 2017 it was estimated that around 1.4 million
people were infected with HIV in the country.
 Tanzania mainland is experiencing a generalized HIV epidemic,
with an HIV prevalence of 4.7% in general population.
 History and discovery of HIV
• When HIV first began infecting humans in 1970s,
scientists were unaware of its existence.
• In early 1980s, new and unusual diagnostic patterns
began to emerge in different parts of the world.
• A benign, fairly harmless cancer called Kaposi’s
Sarcoma, common among the elderly, started
appearing as virulent strain in Younger patients.
• Simultaneously, a rare aggressive form of pneumonia
began to crop up with alarming frequency in another
group of patients in a way specialists had never seen.
• 1981 CDC began to investigate the epidemic and by
the end of the year the first case of AIDS was
documented.
• First cases were discovered from the homosexual
men of Los Angeles.
• Thus in 1982, scientists labeled the group of
mystery illness as Gay-related immune deficiency,
gay cancer or community acquired immune
dysfunction.
• The discovery of the AIDs was published by the
Journal of science by two researchers Luc
Montagnier at Pasteur Institute in Paris (May
20,1983) and Robert Gallo of the National Cancer
institute in Bethesda Maryland May 4 1984.
• They both described the retrovirus obtained
from the cultured T lymphocytes derived from
the lymph nodes of the HIV patient.
• Hence the name Lymphadenopathy- Associated
Virus.
• Patient zero is the first patient to be infected with
HIV and is most likely the one resulting to the spread
of the disease.
• Randy Shilts an author suggested that a Canadian
homosexual flight attendant was a patient Zero, 1984.
• The Attendant admitted to have had 2500 unprotected
unsafe sex partners even after developing Kaposi
Sarcoma.
• This theory was disapproved due to other cases
discovered as early as 1959, Kinshasa DRC preserved
blood sample positive for HIV.
• At the end of 1986 and the beginning of 1987, the
U.S Food and drug administration (FDA)
administered a clinical trial of
Azidothymidine(AZT) the first drug to prove
effective against the rapidly replicating HIV virus.
• By 1993, over 2.5 million cases of HIV/AIDS had
been confirmed worldwide.
• By 1995, AIDS was the leading cause of death for
Americans aged 25 to 44.
• Former Los Angeles basketballer and Lakers player
Earvin (Magic) Johnson announced a press
conference 1991 that he had HIV.
• As soon as he had diagnosed with HIV he began
antiretroviral therapy under the Laker’s physician.
• His public announcement of his HIV-positive status
in 1991 helped dispel the stereotype, still widely
held at the time, that HIV was a "gay disease" that
heterosexuals need not worry about; his bravery in
making this announcement was widely commended.
Magic Johnson
 Origin of the virus
• The most widely accepted is the Hunter theory or
extensions of it.
• Commonly accepted theory, HIV is a viral zoonosis.
• A virus related to it has been isolated from a
chimpanzee native to Cameron, Gabon, DRC
• The virus is called SIV(Simian)
• The chimpanzee is the natural reservoir of the HIV
virus.
• The chimpanzees were hunted and eaten and the
blood got into contact with the hunters who then
were infected.
Other theories include;
 House cat theory (Felin immunodeficiency virus)
1987
 The contaminated vaccine syringes in 1950s
 The contaminated chat vaccine for polio from kidney
of the infected chimpanzee
 The colonization theory
 The conspiracy theory
 If interested read about other theories
 Retroviruses
• This is a family of single stranded RNA viruses
containing enzyme Reverse transcriptase that
allows for reversal of Genetic transcription
from RNA to DNA rather than the usual DNA
to RNA.
• Retro = Reversal
 Etiology of HIV
• AIDS is caused by a retrovirus, namely human
immunodeficiency virus (HIV), belonging to
lentivirus family of Retroviruses.
• On entry into the host cell, they transcribe DNA
which is a complementary copy of RNA.
• The DNA, in turn is used, as a template to produce
new viral RNA copies.
 Types of HIV viruses
• Two different forms of HIV, namely HIV-1 and
HIV-2 have been isolated from AIDS patients.
• HIV-1 is more common, being found in AIDS
patients of USA, Canada, Europe and Central
Africa while HIV-2 is mainly found in West
Africa.
• Both the viruses are almost similar except they
differ in certain immunological properties.
• In Tanzania, HIV infection is caused by HIV-1
sub-types.
• The common HIV-1 sub-types (clades) in
Tanzania are A, C, D, and their recombinants.
• There is no HIV-2 sub-type infection has been
reported up to date.
 Structure of HIV

• The virus is spherical

with a diameter of
about 110 nm.
• The outer shell of the virus
is known as the Viral
envelope.
• This is made of lipid
bilayer that is derived from
the host plasma membrane.
• Embedded in the viral
envelope is a complex
protein gp 160 known as
Cap (gp 120)
env which consists of an
outer protruding cap
Stem (gp 41)
glycoprotein (gp) 120, and a
stem gp 41.
• Within the viral envelope
is an HIV protein called
p17(matrix), found
underneath the bilipid
layer.
• And within the Matrix it
is is the viral core or
capsid, which is made of
another viral protein
p24(core antigen).
Capsid P24
• The viral capsid is
Icosahedral (20 sided)in
shape.
• The capsid protein ,
p24, covers the two
copies of the + single
stranded RNA
(+ssRNA)
• Reverse transcriptase
(p64) and an integrase
(p32) are bound in viral
genome
• A non bound protease,
p10 found in the
nucleocapsid.
 Genome and gene products of HIV
 The HIV genome contains 3 structural genes:
1. env
2. gag
3. pol

 On either side of the HIV genome are long

terminal repeat (LTR) genes which control
transcription.
 1. env gene
• The env gene codes for the envelope precursor gp 160,
which is cloven into gp120 and gp41.
• gp120 forms the 72 knobs which protrude from outer
envelope.
• gp41 is a transmembrane glycoprotein antigen that spans
the inner and outer membranes and attaches to gp120.
• gp120 and gp41 are involved in attachment and fusion of
HIV to CD4 antigen on host cells.
• The surface antigen gp120 is very important for the viral
infection and the detection of AIDS.
gp 160
 2. gag gene

• The gag gene codes for core proteins(capsid

proteins).
• The gag precursor protein p55, which is cleaved by
viral protease to generate p24(capsid protein),
p17(matrix protein), p9 and p6 gag proteins.
 3. pol gene
• Codes for the pol precursor, which is cloven into viral
enzymes reverse transcriptase (RT), protease (PR), and
integrase (IN).
• Reverse transcriptase is responsible for conversion of viral
RNA into DNA into the host cell chromosomal DNA.
• Integrase is involved in the integration of the proviral DNA
into the host DNA.
• Protease is responsible for cleavage of protein
precursors.
Regulatory genes
 Besides the structural genes, HIV contains several
regulatory genes including vif, vpr, tat, rev, vpu and
nef.
 These genes control the synthesis and assembly of
infectious viral proteins.
 In fact, the regulatory genes of HIV play a key role in
the development of AIDS.
• In the absence of Rev, no structural proteins are
made.
Genome and gene products of HIV virus
 Immunological abnormalities in AIDS
 As is evident from the name AIDS, immunodeficiency
(or immunosuppression) is the hallmark of this
disease.
 AIDS primarily affects the cell-mediated immune
system which protects the body from intracellular
parasites such as viruses, protozoa and mycobacterium.
 The primary site for attachment of gp 120 is the
(cluster determinant antigen 4) CD4 molecule.
 Thus HIV virus attack the CD4 positive cells.
• CD4 cells may be regarded as master cells of
cell mediated immunity.
• They produce cytokines, macrophage
chemotactic factors, hemopoietic growth
factors, and others involved in the body
immunity.
• Thus reduction in CD4 (cluster determinant
antigen 4) cells results in
immunosuppression manifested in AIDS
 HIV infects CD4+ lymphocytes and will lyse them
during a productive infection.
 Other cells harbor, replicate and bud HIV without
lysis such as;
• Natural killer cells
• CD8+ killer T cells
• Macrophages
• Cells of the nervous system (Astrocytes, neurons,
glial cells and brain macrophages)
• Dendritic cells
• The major cellular receptor is a CD4 that is
abundantly present on the T lymphocytes.
• Besides CD4 molecules, other two co-
receptors are involved in attachment of HIV
virus to the host cell. This is CCR5
(chemokine Receptor #5) and CXCR4.
• The macrophages corresponds to CCR5
• The T lymphocytes co receptor is CXCR4 (also
known as Fusin)
• When the HIV virus attaches to a cell , the gp120
portion of the virus interacts with CD4.
• Subsequently gp 120 and gp 41under go a
conformational change to attach the CCR5 and
CXCR4.
• The rapid progression of AIDS has been associated
with a switch in co receptor preference from CCR5
to CXCR4.
• During the early course of infection, HIV is a
macrophage tropic.
• In the later stages HIV is T lymphocyte tropic.
• In early phase HIV
infection, initial viruses
are M-tropic.
• Their envelope
glycoprotein gp120 is
able to bind to CD4
molecules and
chemokine receptors
called CCR5 found on
macrophages
• In late phase HIV
infection, most of the
viruses are T-tropic,
having gp120
capable of binding to
CD4 and CXCR4
found on T4-
lymphocytes.
VIRUS LIFE CYCLE
A. The virus through its envelope proteins
attaches to the CD4 receptor and co-
receptors found on the surface of T
lymphocytes and macrophage to gain entry to
the host cells.
B. The virus and cell membrane fuse, and the
virion core enters the cell.
C. The viral RNA and core proteins are
released from the virion core and are then
actively transported to the nucleus.
D. The viral RNA genome is converted into double-
stranded DNA through an enzyme unique to
viruses, reverse transcriptase (red dot).
E. The double-stranded viral DNA moves into the cell
nucleus
F. Using a unique viral enzyme called integrase, the
viral DNA is integrated into the cellular DNA.
G. Viral RNA is synthesized by the cellular enzyme
RNA polymerase II using integrated viral DNA as a
template. Two types of RNA transcripts shorter
spliced RNA (H) and full-length genomic RNA (J)
are produced.
H. Shorter spliced RNAs are transported to the
cytoplasm and used for the production of
several viral proteins that are then modified in
the Golgi apparatus of the cell(I).
J. Full-length genomic RNAs are transported to
the cytoplasm (K).
L. New virion is assembled and then buds off.
M. Mature virus is released.
Life cycle of HIV
 Viral Replication
• The gp120 protein on virus binds specifically to CD4
receptor on host cell with high affinity.
• Gp41 causes fusion of the virus to the cell
membrane.
– After fusion virus particle enters cell.
– Viral genome exposed by uncoating particle
• Reverse transcriptase produces viral DNA from
RNA.
– Becomes a provirus which integrates into host DNA.
– Period of latency occurs.
• After a period of latency lasting up to 10 years
viral replication is triggered and occurs at high
rate.
• CD4 cell may be destroyed in the process,
body attempts to replace lost CD4 cells, but
over the course of many years body is unable
to keep the count at a safe level.
• Destruction of large numbers of CD4 cause
symptoms of HIV to appear with increased
susceptibility to opportunistic infections,
disease and malignancy.
 HIV Transmission
• Transmission of AIDS essentially requires the
exchange of body fluids (semen, vaginal
secretions, blood, milk) containing the virus or
virus-infected cells.
• There are three major routes of HIV transmission;
sexual contact, parenteral inoculation, and from
infected mothers to their newborns.
• The distribution of risk factors for AIDS
transmission are as follows;
Sex between men (homosexuals)- 60%
Sex between men and women – 15%
Intravenous drug abusers — 15%
Transfusion of blood and blood products — 6%
All others — 4%
• More than 90% of adults in sub-Saharan Africa
acquire HIV infection through unprotected
sexual intercourse with infected partners.
• The practice of ‘needle sharing’ is mainly responsible
for the transmission of HIV in drug abusers.
• Pediatric AIDS is mostly caused by vertical
transmission (mother to infant).
• Transmission of HIV is also possible through other
body \such as cerebrospinal fluid, pleural fluid and
amniotic fluids.
• However, unless blood is visibly present, saliva,
sputum, sweat, tears, faeces, nasal secretions, urine,
and vomits carry a very low risk of transmission of
HIV
 NATURAL HISTORY OF HIV
INFECTION
 Classically, the natural history of HIV
infection can be divided into four distinct
stages:
1. Acute primary infection syndrome
2. Asymptomatic latent state
3. Symptomatic HIV infection
4. AIDS.
 Acute primary infection syndrome
• It may be associated with influenza like illness
with fevers, malaise, enlarged lymph nodes,
diarrhea, skin rash, joint pain, fatigue, sore throat
and neurological symptoms such as headache.
• However, up to 60 percent of individuals with
primary HIV infection will be asymptomatic.
• This illness usually lasts 2 to 3 weeks, with full
recovery
• This acute febrile illness is accompanied by
widespread dissemination of the virus to
different tissues, especially the lymphoid
system.
• HIV antibody test often negative but becomes
positive within 3 to 6 months, this process is
known as seroconversion.
Seroconversion — refers to the development of
detectable antibodies against HIV antigens.
• Primary HIV syndrome resolves itself and HIV
infected person remains asymptomatic for a
prolonged period of time, often years.
 Asymptomatic latent state
• Refers to the asymptomatic carrier state that follows initial
infection.
• This stage is free of symptoms, except for the possibility of
swollen glands: Persistent Generalized Lymphadenopathy
(PGL).
• However, this is the stage where there is ongoing extensive
immunologic fighting/changes, rapid viral replication begins
and there is gradual decline in the number of circulating CD4+
T cells.
• This may last for an average of eight to ten years.
• This is WHO Stage 1.
 Symptomatic HIV
• Over time, the immune system loses the struggle to
contain HIV, resulting in extensive destruction of CD4
cells.
• This is characterized by the occurrence of opportunistic
infections (OIs), which is when symptoms develop.
• The most common symptoms include fever, respiratory
infections, cough, TB tuberculosis, weight loss, skin
diseases, viral infections, oral thrush, pain, and
Lymphadenopathy.
• This is WHO Stage 2 or 3, depending on the particular OI
seen.
 Oral Candidiasis (thrush)

Kaposi’s sarcoma is a rare cancer

of the blood vessels caused by
human herpesvirus 8(HHV-8)
that is associated with HIV. It
manifests as bluish-red oval-
shaped patches
 Acquired Immune Deficiency Syndrome (AIDS)

•  AIDS is the outcome of chronic HIV infection and consequent

depletion of CD4 cells.
• It is defined as a CD4 cell count <200 cells/microL or the
presence of any AIDS-defining condition such as Kaposi’s
sarcoma, Cryptococcus meningitis, PCP, toxoplasmosis, CMV
(Cytomegalovirus), cervical cancer etc. regardless of the CD4 cell
count.
• This stage is characterized by severe immunosuppression. This is
WHO Stage 4.
• Note: The risk of HIV transmission is higher in primary HIV
infection and AIDS when the viral load is higher in the blood
WHO Clinical Staging of HIV Disease in Adults and Adolescents
 Stage 4 cont..

HIV wasting syndrome

 Diagnosis of HIV infection
• The diagnosis of HIV infection depends upon the
demonstration of antibodies to HIV and/or the direct
detection of HIV or one of its components.
• The following laboratory tests are employed to diagnose
the HIV infection.
1. ELISA (enzyme-linked immunosorbant assay), detects
antibodies in the circulation.
2. Western blot technique, a more specific test for the HIV
antibodies, is employed for confirmation of ELISA
positive cases.
3. PCR can be used to detect the presence of the HIV
genome in the peripheral blood lymphocytes.
Types of HIV diagnostic tests
– HIV Antibody tests: serology
– Viral antigen test: P24
– Viral nucleic acid
– Virus isolation and culture
 HIV RAPID TESTS

• Rapid diagnostic tests

can be used in adults
and children older than
18 month to detect the
HIV antibodies in blood
or saliva.
 ELISA (enzyme-linked immunosorbant assay)

• First serological test developed to detect HIV

infection.
• In response to the HIV infections the body
develops antibodies directed towards specific
HIV antigens.
• Antibodies detected in ELISA include those
directed against: p24, gp120, gp160 and gp41.
• Different types of ELISA techniques used:
– indirect
– competitive
– sandwich
• ELISAs are for screening only, false positives
do occur and may be due to alcoholism,
syphilis, and immunoproliferative diseases.
• Positive ELISA test need to be confirmed
using Western blot technique.
ELISA (enzyme-linked immunosorbant assay)
 Western Blot
• Most popular confirmatory test.
• It Utilizes a lysate (fluid containing the contents of
lysed cells) prepared from HIV virus.
• The lysate is electrophoresed to separate out the HIV
proteins (antigens).
• The paper is cut into strips and reacted with test sera.
• After incubation and washing anti-antibody tagged with
radioisotope or enzyme is added.
• Specific bands form where antibody has reacted with
different antigens.
• The following antigens must be present: p17, p24, p31,
gp41, p51, p55, p66, gp120 and gp160.
Lysate containing HIV protein is made from the blood
sample
This is then electrophoresed to separate out the HIV
antigen
The formed sera(containing HIV antigen is placed on the
membrane
•The antibody or enzyme is added on the membrane containing the
antigen
•This result in formation of bands were antibody reacts with
different antigen
•The formation of bands signifies presence of antigens; p17, p24,
p31, gp41, p51, p55, p66, gp120 and gp160 antigen
• Interpretation of
results.
– No bands, negative.
– In order to be interpreted
as positive a minimum of
3 bands directed against
the following antigens
must be present: p24, p31,
gp41 or gp120/160.
• CDC criteria require 2
bands of the following:
p24, gp41 or gp120/160.
 Polymerase Chain Reaction (PCR)
• Looks for HIV DNA in the WBCs of a person.
• PCR amplifies tiny quantities of the HIV DNA
present, each cycle of PCR results in doubling
of the DNA sequences present.
• HIV DNA polymerase chain reaction (PCR)
method is used to confirm HIV infection in
infants and children ≤18 months of age.
 Antiretroviral therapy (ART)
Types of Antiretroviral Drugs
 The antiretroviral drugs mainly used in Tanzania fall
into the following five main categories:
a. Nucleotide reverse transcriptase inhibitors (NtRTIs)
b. Nucleoside reverse transcriptase inhibitors (NRTIs)
c. Non-nucleoside reverse transcriptase inhibitors
(NNRTIs)
d. Protease inhibitors (Pls)
e. Integrase strand transfer inhibitors (INSTI)/ Integrase
inhibitors
 Other antiretroviral drugs used elsewhere
include:
f) Fusion inhibitors e.g. Enfuvirtide (ENF)
g) Chemokine receptor inhibitors/ CCR5
inhibitors e.g. Maravir
 Type of Antiretroviral Mechanism of action
Drug
Nucleotide Reverse Selectively inhibiting viral
Transcriptase Inhibitors reverse transcriptase
(NtRTIs) enzyme
Nucleoside Reverse inhibition of viral RNA-
Transcriptase Inhibitors dependent DNA
(NRTIs) polymerase (reverse
transcriptase) enzyme.
Non-Nucleoside Reverse Similar to the NRTIs,
Transcriptase Inhibitors NNRTIs also act by
(NNRTIs) disrupting the reverse
transcription of viral RNA
into DNA that is then
incorporated in the cell’s
nucleus
Examples
•Tenofovir disoproxil
fumarate (TDF)
•Tenofovir alafenamide
(TAF)
•Zidovudine (AZT)
• Lamivudine (3TC)
•Emtricitabine (FTC)
•Abacavir (ABC)
1st generation NNRTIs
•Nevirapine (NVP)
•Efavirenz (EFV)
2nd generation NNRTIs
•Etravirine (ETR
 Type of Mechanism of action
Antiretroviral Drug
Protease Inhibitors (PIs) competitively inhibit the HIV
protease enzyme whose
activity is critical for the
terminal maturation of
infectious virions
Integrase inhibitors This group of drugs acts by
inhibiting integrase enzyme
which facilitates integration
of viral pro-DNA into the
host cell.
Examples
1st generation (PIs)
•Atazanavir (ATV).
• Lopinavir (LPV),
• Ritonavir (usually used as a
booster with other PIs)
2nd generation PIs)
•Darunavir (DRV)
•Dolutegravir (DTG)
• Raltegravir (RAL)
 Prevention
• Abstinence from sex
• Be faithful
• Condom
• Screening during Antenatal period to prevent
MTCT
• Pre exposure prophylaxis (PREP)
• Post exposure prophylaxis (PEP)
 Vaccine against AIDS
• Failure so far
• Why???
• Lack of proof-reading activity by the enzyme reverse
transcriptase. This leads to very frequent alterations
in the DNA base sequence synthesized from viral
RNA which, in turn, influences the amino acid
sequence of proteins. Thus, the protein products of HIV
are highly variable in the amino acid composition and,
therefore, the antigenic properties. For this reason, it
has not been possible to develop a vaccine against
AIDS.
 Summary
 AIDS is a retroviral disease caused by human immunodeficiency
virus (HIV).
 HIV enters CD4 T-lymphocytes where its genetic material RNA is
transcribed into DNA by the enzyme reverse transcriptase. The viral
DNA gets incorporated into the host genome ultimately leading to
the multiplication of the virus and the destruction of CD4 cells.
 This is the root cause of immunosuppression leading to
opportunistic infections in AIDS.
 The sensitive laboratory tests for AIDS detection are ELISA, Western
blot technique and, recently PCR.
 There is no cure and vaccine for AIDS so far however the
administration of ARTs which acts prolongs the life of AIDS
patients.
 REFERENCES
• Textbook of Medical Biochemistry 8th Edition
by Dr. MN Chatterjea
• Biochemistry, 4e by Dr. U. Satyanarayana and
Dr. U. Chakrapan
• National Guideline for the Management of
HIV AIDS by MOH
THANK YOU FOR YOUR ATTENTION