Retroviruses

Retroviruses
 Probably the most studied group of viruses in
molecular biology!!!
 Enveloped, positive-strand RNA viruses
 Unique morphology and replication
 Replicate through a DNA intermediate by
reverse transcriptase (RT)
 Retroviruses
 Baltimore and Temin in 1970
 RNA-dependent DNA polymerase (reverse
transcriptase ) encoded by retroviruses
 Retroviruses replicate through an DNA
intermediate
 This DNA copy of viral genome integrates into
host chromosome
 This discovery earned the Nobel prize: contradicted
the central dogma of molecular biology-genetic
information passed from DNA to RNA and
then to protein
 Here: from RNA to DNA
 History
 Rous sarcoma virus: solid tumors in chicken
 Other cancer causing retroviruses from
other animal species (oncogenes)
 1981: first human retrovirus: Human T-
lymphotropic virus (HTLV-1)
 1983: Human immunodeficiency virus
(HIV)
Subfamily Examples

Oncovirinae
B Mouse mammary tumor virus
C HTLV-I,HTLV-II, Rous sarcoma
virus
D
Lentivirinae HIV-1,HIV-2
Spumavirinae Human foamy virus
Endogenous viruses Human placental virus
 Oncoviruses:immortalize or transform target cells, A,B,C,D
type according to their core and capsid
 Lentiviruses:slow viruses associated with neurologic and
immunosuppresive disease
 Spumaviruses:no disease
 Endogenous viruses:transmitted vertically, 1% of
human chromosome, in many animal species and humans, one
detected in placental tissue which facilitates placental function
 Retroviruses
 Enveloped sperical virion
 Two copies of positive-strand RNA genome
 RT
 Provirus integrates randomly into host chromosome
 Transcription of the genome is regulated by the
interaction of host transcription factors with promoter
and enchancer elements in the long-terminal repeat
portion (LTR) of the genome
 Retroviruses
 Simple retroviruses encode gag,pol and env genes
 Complex viruses also encode accessory –regulatory
genes (tat,rev,nef, vif, vpu for HIV)
 Assembles and buds from the plasma membrane
 Final morphogenesis requires protease cleavage of gag
and gag-pol polypeptides after envelopment.
HIV genome
 Gp120
 CD4 surface receptor protein
 Initially expressed on cells of the macrophage
lineage (macrophage, dendritic cells, microglial
cells) (M-tropic)+ second receptor CCR5
 Later on helper T cells (T-tropic) +fusin
(CXCR4)
 Chemokine receptors
 CCR5: binds macrophage-tropic, non-syncytium-inducing (R5)
viruses  mucosal and intravenous transmission of HIV
infection.

 CXCR4: T-cell-tropic, syncytium-inducing (X4) viruses, which

are frequently found during the later stages of disease.

 In up to 13% of individuals of northern European descent, a

naturally occurring deletion of 32 base pairs in the CCR5 gene
results in a mutant CCR5 receptor that never reaches the cell
surface. Individuals homozygous for this mutation (1-2% of the
Caucasian population) are almost completely resistant to HIV
infection.
 Transmission
*Blood, semen,vaginal secretions
 Sexual contact

 Exposure to contaminated blood and blood

products
 From infected mother to her baby perinatally
 HIV is not transmitted
 Casual contact
 Touching, hugging, kissing, coughing, sneezing,
insect bites, water, food, utensils, toilets,
swimming pools, public baths
 Transmission
 Inoculation of blood: Transfusion,
needlesharing among intravenous drug abusers,
needlestick, open wound, mucous membrane
exposure, tattoo needles
 Sexual trasmission: anal and vaginal intercourse
 Perinatal transmission: Intrauterine, peripartum
and breast milk
 Population at high risk
 Intravenous drug abusers, sexually active people
with many partners (homosexual, heterosexual),
prostitutes, newborns of HIV infected mothers
 Blood and organ recipients and hemophiliacs:
before 1985 (pre-screening programs)
 Epidemiology
 Late 1970s-early1980s
 Young homosexual men, Haitians, heroin addicts,
hemophiliacs were noted to be dying of normally
benign opportunistic infections
 HIV appears to have evolved since 1930s from a simian
virus and then rapidly spread Africa and the world by
an increasinly mobile population.
 There is an expanding epidemic worldwide.
 AIDS cause by HIV
 Retroviridae family
 Lentivirinae genus
 Enveloped, positive strand RNA virus
 2 identical 9-10kb RNA
 AIDS
 Human immunodeficiency virus
type 1 and 2
(HIV-1, HIV-2)
 HIV
 HIV-1: isolated in1983
 Responsible from AIDS pandemic
 HIV-2: isolated in 1986
 HIV-2 less pathogenic
slow progression to AIDS
 HIV group and subtypes

 Rapid mutation and recombination

HIV-1
 Group M (major): A-J
 Group O
 Group N
HIV-2
 A-E subtypes
 Acute retroviral syndrome
 First signs occur in days to several weeks
 Transient
 %50-70
 Activation of immune system
 Multisystem dysfunction
 Flu or infectious mononucleosis sydrome like findings
 Then a latent period
 AIDS
 Continuous viral replication
 Immune system dysfunction
 CD4 T lymphocyes
 > 500 /µl (> %29) 1
 200-499 (% 14-28) 2
 < 200 /µl (< %14) 3
 Incubation

 Adults with no treatment: 10-11 years

 ‘rapid progressors’ : 2-3 years

 ‘non-progressor’ : 7-10 years Stable

CD4 cell count
 HIV genome
 Structural genes: gag/env
 Pol: enzymes -reverse transcriptase

-protease
-integrase
Tat and Rev: necessary for replication
Accessory genes: Vif, Vpr(HIV-1), Vpu, Nef
Vpx(HIV-2)
 HIV
 Viral RNA (in free Virion )
 Viral DNA: integrated in host cell DNA
(Provirus)
 Seroconversion
 Usually 3 weeks
 1.5,3,6,12 months
Viral Kinetics
 Laboratory diagnosis
Serology: Adults and children older than 15 months:
 Initial screening: ELISA, latex agglutination

 Confirmation: Western-blot

Molecular techniques:
-qualitative DNA detection: babies younger than 15
months
-quantitative RNA: follow up of HIV infected people
who are on therapy
Western blot (WB)
 Other tests
 Immunologic status: CD4:CD8 ratio Low
 Antiretroviral resistance tests
 Indicators of disease
Opportunistic infections:
 Protozoal:Toxoplasmosis

 Fungal:Candidiasis, Pneumocystis carinii

 Viral :Cytomegalovirus, HSV

 Bacterial:Mycobacterium avium-intracellulare

Opportunistic neoplasias: Kaposi’s sarcoma

 CCR5 Antagonist
Maravirıoc (MVC) – Selzentry
 Integrase inhibitor

Raltegravir (RAL)/ Isentress

 Prevention and control
 Education
 Blood and blood product screening
 Infection control:
‘Universal Blood and body fluid precautions’:
All patients may be infectious for HIV and other blood-
born infections:
-wear protective clothing(gloves, masks, gown) and other
barriers to prevent exposure to blood products
 Prevention and control
 10% household bleach(% 0.5 chlorine,
5g/litre
5000ppm

 70% ethanol
 2% glutaraldehyde
 4% formaldehyde
 6% Hydrogen peroxide
 Washing laundry in hot water with detergent is
sufficient to inactivate HIV.
 Modes of control
 Antiviral drugs limit progression of disease
 Vaccines for prevention and treatment are in trials
 Safe(Condom!) monogamous sex helps limit spread.
 Sterile injection neeedles should be used
 Large scale screening programs for blood transfusion,
organs for transplants, clotting factors
 Oncovirinae
 RNA tumor viruses
 Associated with leukemias, sarcomas and
lymphomas in many animals
 Not cytolytic
 Distinquished by the mechanism of cell
transformation and length of latency period
between infection and the development of
disease
 Oncovirinae
Sarcoma and acute leukemia viruses:
 Protooncogenes(at least 35)
 Highly oncogenic, direct effect
 No human virus
Leukemia viruses:
 No oncogene
 Long latency period
 HTLV-I,HTLV-II,HTLV-5
 Human T lymphotropic virus
type 1( HTLV-I)
 Adult acute T-cell lymphocytic leukemia (ATLL)
 HTLV-associated myelopathy (tropical spastic
paraparesis)
 Blood transfusion, sexual intercourse, breast
feeding
 Long latency period: approximately 30 years
 Human T lymphotropic virus
type 1( HTLV-I)
 Endemic in southern Japan
 ATLL (1 in 20 people over a 30-50 years
 Diagnosis: ELISA+WB
 Viral RNA by RT-PCR
 HTLV-2: Hairy cell leukemia
 HTLV-5: malignant cutaneous lymphoma