Notions de virologie

Table des matières

• Morphologies de virus
• Qu’est ce qu’un virus
• Cycle de vie
• Organisation génétique des virus
• Virus et santé : l’exemple du HIV
 Virus hélicaux
• Tube spiralé de protéines (“colimaçon”)
• Rage, virus mosaïque du tabac
 Virus icosahédriques
• Semblent ronds au microscope
• 20 faces triangulaires
• Hépatite B
 Virus enveloppés
• Sphériques
• Ces virus utilisent la membrane de l’hôte
pour former une enveloppe
• Des Glycoprotéines (“récepteurs”) à la
surface du virus lui permettent de ne pas
être reconnu
• HIV, grippe
Virus enveloppés
 Bactériophage

• Tête hexagonale et queue en hélice

• La queue sert à injecter l’ADN du virus

dans la bactérie hôte

• Phage T4
Bactériophage
 Caractéristiques

• Petite taille: 20-300 nm de diamètre

• Entièrement dépendant d’une cellule hôte pour sa
survie
• Certains virus codent néanmoins pour une ADN
polymérase ou une ARN polymérase, mais ne peuvent
jamais se multiplier seuls
• Récepteurs de surfaces pour la reconnaissance et
l’adhésion à la surface hôte
• Vecteurs très important de transfert horizontal entre
espèces
Taille des virus
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 Réplication et cycle de vie

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 Une grande variété de mécanismes d’entrée

• Certains virus entrent également dans la cellule par ce

moyen, mais ne peuvent pas fusionner leur membrane avec
celle de l’endosome

•Poliovirus: l’endosome est acidifié et expose les virions sur

la membrane, d’où ils vont ensuite être relachés dans le
cytoplasme

•Reovirus: transfert du contenu de l’endosome au

lysosome, où les protéases vont détruire la capside virale et
laisser l’ADN s’échapper
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Poliovirus

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Mécanismes
d’exocytose

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 Génétique virale

• Le matériel génétique existe sous forme d’ADN ou d’ARN, simple brin ou

double brin
• Il peut être circulaire ou linéaire
• Le génome peut etre divisé en plusieurs chromosomes, . e.g. le virus
influenza et les rotavirus
Génomes viraux
Expression génétique coordonnée
 Evolution des virus
• Le nombre de générations est très grand, ainsi que la
population, le nombre potentiel de mutations est
donc très important, même sur des temps courts

• L’évolution précise dépend beaucoup de la structure

du génome (ADN ou ARN, segmenté ou non)

• L’un des principaux mécanismes d’évolution virale

répond à une forte pression de sélection pour
échapper au système immunitaire de l’hôte: on
parlera de shift antigénique
 Mutations et recombinaison

• Des mutations ont lieu naturellement lors de

la réplication du virus par la cellule hôte (cf les
virus qui ont leur propre ADN polymérase « à
erreurs »)
• Les mutations sont plus fréquentes chez les
virus à ARN
• Des mécanismes de recombinaison peuvent
avoir lieu dans la population virale au sein
d’un hôte, ou entre les segments d’un génome
Le virus influenza (la grippe)
 Quels traitements?
• Les antibiotiques ne fonctionnent pas
• Prévention : vaccins viraux
• Les cellules immunitaires produisent des
interférons (un type de cytokine) pour se
défendre contre les virus
• Thérapies antivirales à base d’analogues de
nucléosides ou de nucléotides, d’inhibiteurs
de la reverse-transcriptase ou des protéases
 Vaccins viraux
• L’idée est d’injecter à la personne un virus
modifié, qui n’est pas pathogène, mais dont la
présentation des antigènes va activer le
système immunitaire
• On peut imaginer généraliser ce système avec
des peptides de synthèse, des virus vivants
atténués
• Thérapie génique?
 Interférons
• Produits naturellement par des cellules
immunitaires en contact avec des virus ou des
cellules infectées
• Empêchent la réplication virale, stoppant
l’infection
• La défense naturelle principale contre les virus
L’exemple du HIV
Cycle de vie du HIV-1
Organisation génétique du HIV-1
Une histoire évolutive complexe
 CHARACTERISTICS

• To see the virus electron microscope

• Growth need living cells/ tissues
• Can not growth saprophytic
• Only have certain enzyme for metabolism and energy
• Easy mutated changes antigenic property
• Multiplication different from bacteria

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 Virus structure & Morphology

The basic design of all viruses places the nucleic

acid genome on the inside of a protein shell
à capsid

Two basic types of virions :

1. Enveloped viruses
à have a nucleocapsid of nucleic acid
complexed to protein
2. Naked capsid viruses
à have a nucleic acid genome within a
protein shell

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 Virus structure & Morphology

Schematic drawing of two basic type of virions

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 Two basic shapes of virions :
1. Cylindrical
2. Spherical
Some bacteriophages combine those 2 basic shapes

Functions of capsid or envelope of viruses :

1. To protect the NA genome from damage during extra--
extra
celullar passage of the virus from one cell to another
2. To aid in the process of entry into the cell
3. To package enzymes essential for the early steps of the infection
process

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Basic viral
forms

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 The structure
and relative
sizes of a
number of
DNA

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 The structure
and relative
sizes of a
number of
RNA

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 DNA VIRUSES

ENVELOPED NAKED

Double – stranded Double – stranded Single

Single--stranded

Icosahedral Complex Icosahedral Icosahedral

HERPES POX PAPOVA PARVO

HEPADNA ADENO

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 RNA VIRUSES
Single – stranded Double – stranded

Positive – stranded (+) Negative – stranded (-)

Naked Enveloped Enveloped Naked

*PICORNA *TOGA BUNYA *REO

*CALICI *FLAVI ORTHOMYXO
CORONA PARAMYXO
RETRO RHABDO
ARENA
FILO
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* Icosahedral; all of the rest have helical symmetry
 ADSORPTION
Adsorption is the first step in every viral infection.

Adsorption involves :
- virion attachment proteins
- cell surface receptor proteins

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Examples of viral receptors
• For some viruses co-
co-receptors are involved in adsorption
à HIV
HIV--1 : CD4
CD4 & chemokine receptors

• Viral spikes & phage tails carry attachment proteins

• In some case, a region of the capsid protein serve

the function of attachment

• Adsorption is enhanced by presence of multiple

attachment & receptor proteins.

• A particular kind of virus is capable to infecting only

a limited spectrum of cell types à its host range
Differences in host range & tissue tropism due to
presence or absence of the receptors

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 Entry & Uncoating

Enveloped Animal Viruses

Some enveloped viruses enter cells by direct fusion
of plasma membrane & envelope, release the
nucleocapsid directly into the cytoplasm.
à Paramyxoviruses (eg. measles) retroviruses
(eg. HIV-1) & herpesviruses

Other enveloped & naked viruses are taken in by

receptor--mediated endocytosis (viropexis).
receptor
à orthomycovirus (eg. influenza viruses),
togaviruses (eg. rubella viruses), rhabdoviruses
(eg. rabies) & coronaviruses

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 Entry by
direct fusion

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 Viropexis

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Viral release by Budding

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 Late Transcription

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 Human Viral Diseases
• Because viruses aren’t “alive” they must be
spread by an intermediate host (vector)
 Chicken Pox/Shingles:
• Caused by vericella-zoster herpesvirus
• Spread through the air and contact
• May remain dormant as a provirus and
become shingles
 Viral Hepatitis
• Inflammation of the liver caused by 5
different viruses
• A and E spread by fecal matter
• B,C and D spread by sexual contact or
blood transfusion
 AIDS
• Acquired Immune Deficiency
Syndrome
• Caused by the HIV virus
• Glycoproteins on the surface of the
virus bind to receptor sites of immune
cells (macrophages)
• Retrovirus, Provirus
• Genetically diverse
 Is there a cure?
• Azydothymidine- inhibits reverse
transcriptase
• Protease inibitors- blocks synthesis
of new capsids
• These drugs can only slow the
progression to full-blown AIDS
 Emerging Diseases
• Caused by infections in animal
populations
– Rainforest animals-> Ebola
– Hanta virus (pneumonia)-> mice
– SARS-> civet cats
• Epidemic- quickly spreading infectious
disease
• Pandemic- disease spreads over large
regions
• Bubonic plague, Small pox
 Treatment
• Vaccine- harmless version of a virus used
to build an immune response
(microevolution)
– Inactivated virus- not able to replicate
– Attenuated virus- weakened form of a virus; no
ability to cause disease
• Vector control
• Drug therapy- interfere with DNA/RNA
synthesis
– Acyclovir- blocks DNA polymerase
 PATHOGENESIS OF VIRAL DISEASES
Viral pathogenesis :
interaction of viral and host factors
leads to disease production
Virus pathogenic if : can infect and cause signs of
disease of the host
Virus virulent : produce more severe disease
Steps in viral pathogenesis :
• Viral entry & primary replication
• Viral spread and cell tropism
• Cell injury & clinical illness
• Recovery from infection
• Virus shedding SUNARYATI
 Laboratory Diagnosis

• Identification of the virus in cell culture;

• Microscopic identification directly in the specimen;
• Serologic procedures to detect a rise in antibody titer or the presence of Ig M
antibody;
• Detection of viral antigen in blood or body fluids;
• Detection of viral nucleic acids in blood or patients cells.
Identification of The Virus in Cell Culture

The presence of a virus in a patient's specimen can be detected by

cytopathic effect in cell culture but CPE is not specific. A specific
identification of the virus usually involves an antibody based test
as fluorescent antibody, complement fixation or ELISA.
Microscopic identification directly in the specimen
Electron microscopy is the most common method used to study
the morphology of viruses.

• Inclusion bodies, formed by aggregates of many virus particles, can be

seen in either the nucleus or cytoplasm of infected cells.
• Multinucleated giant cells are formed by several viruses e.g. herpes,
respiratory syncytical virus and measles virus.
• Fluorescent antibody staining of cells obtained from the patients or of the
cells infected in culture can provide a rapid specific diagnosis.
• Electron microscopy is not often used in clinical diagnosis but is useful in
the diagnosis of certain virus e.g Ebola (characteristic appearance and are
dangerous to grow in culture).
Serologic Procedures
• The presence of Ig M antibody can be used to diagnose current infection.
• The presence of Ig G antibody cannot be used to diagnose current infection. Rise in
antibody titer that is 4 fold or greater in the convalescent serum sample compared to the
acute sample can be used to make a diagnosis.

Detection of Viral Antigen & Nucleic Acids

• The presence of hepatitis B surface antigen is commonly used in diagnosis.

• The presence of vial DNA or RNA is increasingly becoming the gold standard in viral
diagnosis. Molecular diagnostic procedures have been available since 1970s,when
researchers first began using cloned DNA probe to detect viral nucleic acid. The new
molecular diagnostic methods predicted that nucleic acid tests would rapidly replace
traditional virus detection methods.
 Molecular Diagnostic Methods
• The goal is in the detection of non culturable agents such as human papilloma
virus, human parvovirus,
• Detecting viruses difficult to culture, including enteric adenovirus, some coxsackie
viruses,
• Detecting viruses that are dangerous to culture such as HIV,
• Detecting viruses that are present in low numbers, for example, HIV in antibody
negative patients or CMV in transplanted organs.
• Important when a tiny volume of specimen is available (forensic samples or intra-
ocular fluid specimens).
• Allow laboratory to predict antiviral drug susceptibilities and to detect infections
when viable virus cannot be obtained (latent viral infection or viruses that are
present in immune complexes).
• May also used to differentiate antigenically similar viruses such as adenovirus
types 40 and 41 and to detect viral genotypes that are associated with human
cancers (human papilloma virus).
 Recombination
• Process of intermolecular exchange, of chromosomes combining genetic
information from different sources, typically two genomes of a given
species.
• This kind of break/join recombination is common in DNA viruses or those
RNA viruses which have a DNA phase (retroviruses).
 PREVENTION AND TREATMENT OF VIRAL INFECTIONS
1. VIRAL VACCINES
– Killed-virus vaccines
– Attenuated live-virus vaccines
– Future prospect :
- attenuation of viruses by genetic
mapping
- avirulent viral vectors
- purified proteins produced using
cloned genes
- synthetic peptides
- subunit vaccines
- DNA vaccines SUNARYATI
2. INTERFERONS
IFNs :
Mhost-coded proteins of large cytokine
family
Minhibit viral replication
Mproduced by intact animal or cell culture in
response to viral infection or other
inducers
Mfirst line of defense against viral infection
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3. ANTIVIRAL CHEMOTHERAPY
A. Nucleoside analogs
• Acyclovir &
– Ribavirin
valacyclovir
• Didanosine – Stavudine (d4T)
• Gancyclovir – Trifluridine
• Idoxuridine – Vidarabine
• Lamivudin (3TC) – Zalzitabine (ddC)
– Zidovudine (AZT)
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B. Nucleotide analogs
Cidofovir : active against CMV & HSV
inhibit s viral DNA polymerase
C. Nonnucleoside reverse transcriptase inhibitor
Nevirapine : inhibit reverse transcriptase of HIV
D. Protease inhibitors
Ritonavir, Saquinavir HIV
E. Other types
Amantadine & rimantadine
Foscarnet
Methiasone
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