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Batterjee Medical College
Batterjee Medical College
Basic Virology: Introduction
• Viruses are not cells
• They are not capable of independent replication
• They synthesize neither their own energy nor their
own proteins
• They are too small to be seen in light microscope.
Batterjee Medical College
Basic Virology: Introduction
Comparison of Viruses and Cells
Property Viruses Cells
Type of nucleic acid DNA or RNA but not both DNA and RNA
Proteins Few Many
Lipoprotein membrane Envelopeviruses present
in some
Cell membrane present in all
cells
Ribosomes Absent Present
Mitochondria Absent Present in eukaryotic cells
Enzymes None or few Many
Multiplication by binary
fission or mitosis
No Yes
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Virus Size & Structure
• Viruses range in size from (~20 nm to ~300 nm) .
• Most viruses appear as spheres or rods in the
electron microscope.
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• All viruses have protein coat called capsid that covers genome.
• Capsid is composed of repeating subunits called capsomers virus symmetric appearance used for classification.
• Some viruses, capsid is covered with lipoprotein envelope.
Virus Size & Structure
Capsid
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• It composed of nucleic acid genome & capsid
proteins.
Virus Size & Structure
Nucleocapsid
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• Viral nucleocapsids have : -Spherical (icosahedral) symmetry (enveloped or naked) -Helical symmetry (enveloped).
Virus Size & Structure
Nucleocapsid
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• Viruses contain either DNA or RNA, but not both.
• These DNA & RNA genomes can be either single-stranded or double-stranded.
• Some RNA viruses (e.g. influenza virus & rotavirus), have segmented genome.
• All viruses have one copy of their genome (except retroviruses, which have two copies).
Viral Nucleic Acids
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• It mediate attachment to host cell receptors (determines host & organ specificity of virus).
• Surface proteins are targets of antibody which "neutralizes" (inhibits) viral replication.
• Some viruses produce antigenic variants of their surface proteins evade host defenses.
Internal proteins :
They are DNA or RNA polymerases.
Viral Proteins
Viral surface proteins
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It mediates interaction between viral nucleocapsid
proteins & envelope proteins.
Viral Proteins
• Antibody against one antigenic variant (serotype) will not neutralize different serotype. • Some viruses have one serotype; others have multiple serotypes.
Matrix protein
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• Envelope consists of membrane that contains lipid
derived from host cell & proteins encoded by virus.
• Envelope is acquired as virus exits from cell in
process called budding.
Viral Envelope
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Viral Envelope
• Enveloped viruses :
-Easily inactivated
-transmitted by direct contact
via blood & body fluids.
• Naked viruses
- Survive longer in
environment
- Transmitted by indirect
means e.g. fecal-oral route
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•One virion infects cell & hundreds of progeny
virions are produced within hours.
•This is remarkable amplification & explains rapid
spread of virus from cell to cell.
The eclipse period
It is time when no virus particles are detected within
infected cell & occurs soon after cell is infected.
Viral Growth Curve
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The steps in viral replication are as follows:
1.Attachment
2.Penetration and uncoating.
3.Transcription & Translation
4.Assembly
5.Release
Viral Replication
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• Virus particles can only infect cells possessing
surface “receptors” specific to particular virus
species.
• Virus attached to cell with:
- Capsid (Naked viruses)
- Envelope proteins
(enveloped viruses)
Viral Replication
1-Attachment
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Viral Replication
• Viruses penetrate into cell by means of pinocytosis
(viropexis).
• In enveloped viruses, envelope fuse with cell
membrane, releasing virus into cytoplasm.
2-Penetration
Adsorption of such an enveloped virus to two cells at the same time may result in cell fusion
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Viral Replication
• It involves release of nucleic acid from capsid
• It is activated by cellular enzymes & contribution
from cell membranes (except smallpox virus)
3- Uncoating
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• All DNA viruses genome is double-stranded
(except parvoviruses, single-stranded)
• They use host cell RNA polymerase to synthesize
viral mRNA.
• All DNA viruses replicate in nucleus (except
poxviruses in cytoplasm).
Viral Replication
4- Replication of nucleic acid
DNA viruses
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Viral Replication
•Genome of all RNA viruses is single-stranded
(Except reoviruses, e.g., rotavirus double-
stranded).
• All RNA viruses replicate in cytoplasm, except
retroviruses, influenza virus, & hepatitis D virus,
which require intranuclear step in their replication.
4- Replication of nucleic acid
RNA viruses
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•Viral proteins:
•Early proteins are typically enzymes used in
synthesis of viral components, whereas late
proteins are typically structural proteins of progeny
viruses.
•poliovirus & retroviruses, translate their mRNA into
precursor polyproteins, which must be cleaved by
proteases to produce functional proteins.
Viral Replication
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Assembly and release:
•All enveloped viruses acquire their envelope by
budding through the external cell membrane as they
exit the cell,
•herpesviruses, acquire their envelope by budding
through nuclear membrane.
Viral Replication
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Viral Replication
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Lysogeny
•It is the process by which viral DNA becomes
integrated into host cell DNA, replication stops, and
no progeny virus is made.
• Later, if DNA is damaged by, for example, UV light,
viral DNA is excised from the host cell DNA and
progeny viruses are made.
•The integrated viral DNA is called a prophage.
Viral Replication
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Lysogeny
Bacterial cells carrying a prophage can acquire new
traits, such as the ability to produce exotoxins such
as diphtheria toxin.
Transduction is the process by which viruses carry
genes from one cell to another.
Lysogenic conversion is the term used to indicate
that the cell has acquired a new trait as a result of
the integrated prophage.
Viral Replication
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Viral Replication
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Genetics
• Mutations in the viral genome can produce antigenic variants and drug-resistant variants.
• Mutations can also produce attenuated (weakened) variants that cannot cause disease but retain their antigenicity and are useful in vaccines.
• Temperature-sensitive mutants can replicate at a low (permissive) temperature but not at a high (restrictive) temperature.
• Temperature-sensitive mutants of influenza virus are used in vaccines against this disease.
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Genetics
Reassortment (exchange) of segments of the genome RNA of influenza virus is important in the pathogenesis of the worldwide epidemics caused by this virus.
•Complementation occurs when one virus produces a protein that can be used by another virus.
•A medically important example is hepatitis D virus that uses the surface antigen of hepatitis B virus as its outer coat protein.
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•Phenotypic mixing occurs when two different
viruses infect the same cell and progeny viruses
contain proteins of both parental viruses.
• This can endow the progeny viruses with the ability
to infect cells of species that ordinarily parental
virus could not.
Genetics
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The Infected Patient•Viral infection in the person typically has four stages:- incubation period,-prodromal period- specific-illness period,- recovery period.
•The main portals of entry are the respiratory, gastrointestinal, and genital tracts, but through the skin, across the placenta, and via blood are important as well.
Pathogenesis
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The Infected Patient•Transmission from mother to offspring is called vertical transmission; all other modes of transmission, e.g., fecal–oral, respiratory aerosol, and insect bite, are horizontal transmission.•Transmission can be from human to human or from animal to human. •Most serious viral infection are systemic, i.e., the virus travels from the portal of entry via the blood to various organs. •some are localized to portal of entry as common cold & involves only upper respiratory tract.
Pathogenesis
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Pathogenesis
•The symptoms of viral diseases are usually caused
by death of the infected cells and a consequent loss
of function.
• For example, poliovirus kills neurons, resulting in
paralysis.
Pathogenesis
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Pathogenesis
Immunopathogenesis
•It is the process by which the symptoms of viral
diseases are caused by the immune system rather
than by the killing of cells directly by the virus.
•One type of immunopathogenesis is the killing of
virus-infected cell by the attack of cytotoxic T cells
that recognize viral antigens on the cell surface
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Pathogenesis
•Damage to the liver caused by hepatitis viruses
occurs by this mechanism.
•Another is the formation of virus–antibody
complexes that are deposited in tissues.
•Arthritis associated with parvovirus B19 or rubella
virus infection occurs by this mechanism.
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Pathogenesis
•Viruses can evade host defenses by producing multiple antigens, thereby avoiding inactivation by antibodies.
•and by reducing the synthesis of class I MHC proteins, Thereby decreasing the ability of a cell to present viral antigens and blunting the ability of cytotoxic T cells to kill the virus-infected cells.
•Viruses produce receptors for immune mediators, as IL-1 and TNF, thereby preventing the ability of these mediators to activate antiviral processes
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Carrier state refers to people who produce virus for long periods of time and can serve as a source of infection for others. The carrier state that is associated with hepatitis C virus infection is a medically important example.
Latent infections are those infections that are not producing virus at the present time but can be reactivated at a subsequent time. The latent infections that are associated with herpes simplex virus infection are a important example.
Persistent Viral Infections
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•Slow virus infections refer to those diseases with a long incubation period, often measured in years.
•Some, such as progressive multifocal leukoencephalopathy, are caused by viruses, whereas others, such as Creutzfeldt-Jakob disease, are caused by prions.
•The brain is often the main site of these diseases.
Persistent Viral Infections
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Host Defenses
•Host defenses against viruses fall into two major categories:
(1) Nonspecific, of which the most important are interferons and natural killer cells
(2) Specific, including both humoral and cell-mediated immunity.
• Interferons are an early, first-line defense, whereas humoral immunity and cell-mediated immunity are effective only later because it takes several days to induce the humoral and cell-mediated arms of the immune response
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Nonspecific DefensesInterferons
• Interferons inhibit virus replication by blocking production of viral proteins, by degrading viral mRNA.
• They induce the synthesis of a ribonuclease that specifically cleaves viral mRNA but not cell mRNA.
• Double-stranded RNA viruses are the most potent inducers of interferons.
• Many viruses induce interferons, and many viruses are inhibited by interferons, i.e., neither the induction of interferons nor its action is specific.
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•Interferons act by binding to a receptor on the cell surface that signals the cell to synthesize the ribonuclease and the other antiviral proteins.
•Interferons do not enter the cell and have no effect on extracellular viruses.
•Alpha & beta interferons have a stronger antiviral action than gamma interferon.
•Gamma interferon activates macrophages.
Nonspecific DefensesInterferons
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• Natural killer (NK) cells are lymphocytes that destroy cells infected by many different viruses, i.e., they are nonspecific.
• NK cells do not have an antigen receptor on their surface.
• NK cells recognize and destroy cells that do not display class I MHC proteins on the surface.
• They kill cells by secreting perforins and granzymes.
Nonspecific Defenses
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•Phagocytosis by macrophages and the clearance of
mucus by the cilia of the respiratory tract are also
important defenses.
Nonspecific Defenses
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• Increased corticosteroid levels suppress various
host defenses and predispose to severe viral
infections as disseminated herpesvirus infections.
• Malnutrition predisposes to severe measles
infections in developing countries.
• The very young and the very old have more severe
viral infections.
Nonspecific Defenses
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Specific Defenses
Active immunity to viral infection is mediated by both antibodies and cytotoxic T cells.It can be elicited either by exposure to the virus or by immunization with a viral vaccine.
Passive immunity consists of antibodies preformed in another person or animal.
The duration of active immunity is much longer than (years) that of passive immunity (weeks to a few months).
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Passive immunity is effective immediately, whereas it takes active immunity 7 to 10 days in the primary response (or 3–5 days in the secondary response) to stimulate detectable amounts of antibody.
Herd immunity is the protection of an individual that results from immunity in many other members of the population (the herd) that interrupts transmission of the virus to the individual. Herd immunity can be achieved either by immunization or by natural infection of a sufficiently high percentage of the population.
Specific Defenses
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Identification in Cell Culture
•The presence of a virus in a patient's specimen can
be detected by seeing a "cytopathic effect" (CPE) in
cell culture.
•CPE is not specific, i.e., many viruses cause it.
• A specific identification of the virus usually
involves an antibody-based test as fluorescent
antibody, complement fixation, or ELISA.
Laboratory Diagnosis
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Microscopic IdentificationInclusion bodies•They are formed by aggregates of many virus particles, can be seen in either the nucleus or cytoplasm of infected cells.
•They are not specific. •Two important examples are the nuclear inclusions formed by certain herpesviruses and the cytoplasmic inclusions formed by rabies virus (Negri bodies).
Laboratory Diagnosis
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Multinucleated giant cells
•They are formed by several viruses, notably certain herpesviruses, respiratory syncytial virus & measles virus. •Fluorescent antibody staining of cells obtained from the patient or of 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 viruses, as Ebola virus, that have a characteristic appearance and are dangerous to grow in culture.
Laboratory Diagnosis
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Serologic Procedures•The presence of IgM can be used to diagnose current infection. •The presence of IgG cannot be used to diagnose current infection because the antibody may be due to an infection in the past.•acute and convalescent serum sample should be analyzed. •An antibody titer that is fourfold or greater in the convalescent serum sample compared to the acute sample can be used to make a diagnosis.
Laboratory Diagnosis
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Detection of Viral Antigens & Nucleic Acids
•The presence of viral proteins, such as p24 of HIV and hepatitis B surface antigen, is commonly used in diagnosis.
•The presence of viral DNA or RNA is increasingly becoming the "gold standard" in viral diagnosis.
•Labeled probes are highly specific and rapidly.•Small amounts of viral nucleic acids can be amplified using reverse transcriptase to produce amounts detectable by the probes e.g."viral load" assay of HIV RNA.
Laboratory Diagnosis