VIROLOGY

MCB 4206

Prepared
By

Professor Usman Aliyu Dutsinma

Department of Microbiology
Bayero University, Kano
 VIRUS REPLICATION
As viruses are obligate intracellular
pathogens they cannot replicate
without the machinery and metabolism of
a host cell Although the replicative life
cycle of viruses differs greatly between
species and category of virus, there are
six basic stages that are essential for
viral replication.
• There are two types of replication cycles in
viruses:
1.Lytic cycle and
2.Lysogenic cycle

1. Lytic cycle involves lysing host cell to release newly

assembled viral population (Cell destruction)

2. Lysogenic cycle where the virus remains latent or

dormant incorporating its genome into the host’s genome
as provirus
LYTIC CYCLE
1. Attachment
Viral proteins on the capsid or phospholipid envelope
interact with specific receptors on the host cellular
surface. This specificity determines the host range (tr
opism) of a virus.
Receptor molecules differ for different viruses. e. g.
poliovirus is able to attach only to cells in CNS/ and
intestinal tract of primates, HIV binds to the CD4
receptor on cells of human immune system.
2. Penetration (Entry)
•This follows attachment adsorption
•Viruses, enter host cell via receptor mediated membrane
fusion.
•This is called viral entry and the process is called
pinocytosis or Viropexis.

•It isaccomplished by receptor- mediated endocytosis with

uptake of the ingested virus particles within endosomes.
Three main mechanisms are involved:
1. Translocation of the entire virus particle across the
cytoplasmic membrane of the cell. It must be mediated
by proteins in the virus capsid and specific
membrane receptors.
Translocation of entire virus particles across the cell membrane by cell-
surface receptors
2. Endocytosis of the virus into
intracellular vacuoles this is probablythe
most common mechanism of virus entry
into cells. It does not require any specific
virus proteins (other than those already
utilized for receptor binding) but relies
on the normal formation and
internalization of coated pits at the cell
membrane.
3. Fusion of the virus envelope (only
applicable to enveloped viruses)
with the
cell membrane, either directly at the cell
surface or following endocytosis in a
cytoplasmic vesicle which requires the
presence of a specific fusion protein in the
virus envelope—for example, influenza
haemagglutinin or retrovirus
transmembrane (TM) glycoproteins.
3. Uncoating
•Uncoating occurs concomitantly with or
shortly after penetration.
•At this stage, there is physical separation of the
viral nucleic acid from the outer structural
components of the virion.
•The infectivity in the parental virus is lost at
this point
•Uncoating is done enzymatically(from
lysosome).
•During uncoating, viral capsid is degraded
by viral enzymes or host enzymes, releasing
the viral genomic nucleic acid, which is
integrated in a host cells chromosome.
This is called Prophage.
4. Replication / Transcription
•At his stage, a specific mRNA must be transcribed for
successful expression and duplication of genetic information
contained in the viral genome.
•The mRNA produced serves as replicative intermediate
from the viral genome.
•The process transcription could be carried out by either the
host cell mechanism or a virus-specified enzyme.
•There is viral protein synthesis
•Assembly of viral proteins also takes place
•Viral genome replication takes place
•The virus nucleic acid instructs the host cell to replicate the
virus DNA or RNA
Viral Maturation (Morphogenesis)
•The viral capsid proteins and genomes are present in
multiple copies after the replication process.

•The capsid proteins and genomes are assembled into

new infectious virus particles.

•This is the stage where the virus uses the host cell
component to translate the viral mRNA.

• All the virus specified macromolecules are

Synthesized in a highly’ organized sequences.
•The virus mRNA is translated on cell ribosomes
to produce two types of virus-protein:

(a) The structural proteins which make up the

virus particle.

(b) Non-structural protein, mainly enzymes for

virus genome replications.

This type of protein is not found in the viral

particle.
5. Assembly
•The newly synthesized viral genomes and capsid
Polypeptides assemble together to form progeny
viruses.

•Icosahedral capsid can condense in the absence

of nucleic acid,

•whereas nucleocapsids of viruses with helical

symmetry cannot form without viral RNA.
6. Release
•New virus particles are released from the
host cell either by lysis or by budding
Lysis
•The cell is killed by bursting of its membrane.
•Bursting is achieved when the virus directs the
production of an enzyme which damages the
host cell wall, causing the host cell to swell
and burst.
 Budding
 The newly assembled virus pushes out ("buds") from the
host cell.
• During budding, the HIV envelope also acquires host
membrane proteins and lipid bilayer
 Nonenveloped and complex viruses are released when the
cell lyses or ruptures
 Enveloped viruses are liberated by budding or exocytosis
 Anywhere from 3,000 to 100,000 virions may be released,
depending on the virus
 Entire length of cycle- anywhere from 8 to 36 hours
 VIRUSES AND HOST DEFENCE

HOST DEFENSE AGAINST VIRUSES

SPECIFIC/ NON- SPECIFIC/
ADAPTIVE INNATE
IMMUNE IMMUNE
RESPONSE RESPONSE

• Humoral • Cellular • Interferons(IFN) • Mx NK

Cells
a. B- cells a. T- helper cells a. Leucocyte IFN (IFNÞ) proteins
(Antibody b. Cytotoxic T- b. Fibroblast IFN (IFNß) (Mx genes)
Production)
Lymphocytes c. Immune IFN (IFN y)
Complements
Immune response to Viral Infections

 A number of specific immune effector mechanisms, toget

her with nonspecific defense mechanisms, are called int
o play to eliminate an infecting virus.

1. Innate immune response to viral infection

 Interferon
 A group of proteins produced in response to virus infe
ction which stimulates cells to make proteins that bloc
k viral transcription, and thus protects them from infec
tion.
 Immune response to Viral Infections

 dsRNA produced during viral replication induce the expre

ssion of interferons by the infected cells.

 Monocytes , macrophages & fibroblasts also synthesize i

nterferons .

 Anti-viral activity of interferons (IFNs)

 Virus infected cells produce INF-α;

 INF-α inhibit intracellular replication of viruses

Immune response to Viral Infections

 IFN-α activate NK-cells to kill virus infected cells

 IFNs have no direct effect on extracellular virus
 IFNs act early in viral diseases before antibody
 INFs activity is not specific
 NK cells
 Destroy some virus-infected cells, and are not MHC r
estricted.
 Natural killer cells lyse virally infected cells
Immune response to Viral Infections

Specific immune response

 Humoral immunity
 Anti-viral antibodies produced by B - lymphocyte :
 prevent spread during acute infection.
 protect against reinfection .
 Virus neutralization:- In viraemic infections,
antibodies neutralize virus, preventing its
attachment to receptor sites on susceptible cells
e.g. Poliovirus, mumps, measles, rubella
Antibody production by B - Lymphocytes

Antibodies

B- Lymphocyte
•These antibodies mainly prevent viral infection
by preventing or hindering virus adsorption to
the host cells. This is called ―Neutralization”
•The neutralizing effect of antibodies is strongest
when they react with the receptor-binding sites
on the capsids thereby blocking them, rendering
the virus in capable of combining with the
cellular receptors.
•Humoral immunity is mainly extracellular
Immune response to Viral Infections

In superficial non-viraemic infections (infleunza)

Secretory IgA neutralizes virus infectivity at the
mucous surfaces.

 Antibodies destroy free virus particles directly by:

i- Aggregation of virus and opsonization
ii- Complement mediated lysis

Both mechanisms also act on virus infected cells

Immune response to Viral Infections

 Cell mediated immunity(CMI)

 Cell – mediated immunity is important for control and
clearance of viral infections.

 CMI acts on virus infected cells through:

- Cytotoxic T-cells (CTLs)
- NK cells
- Activated macrophages

 CTLs kill virus infected cells directly after recognition

of viral antigens on cell surface in association with
Major Histocompatibility Complex I (MHC I)
Immune response to Viral Infections

T – Helper cells stimulated by viral antigens release

cytokines. Cytokines attract and activate macrophages
to kill virus infected cells

 Nk-cells destroy virus infected cells early in infection

before appearance of antibodies

 Antibody-dependent cell mediated cytotoxicity (ADCC):

 Antibody binds to virus infected cells such cells are
lysed by NK cells, macrophages and polymorphs
Cellular Immunity (Cell – mediated immunity)
•This involves fighting viral infections to attain immune
defence.
• T lymphocytes (killer cells) recognize virus-infected cells
by the viral antigens on their surfaces and destroy them.
• T-lymphocytes activate phagocytes (e.g. macrophages) to
destroy microbes that have been ingested (applicable to all
microbes).
•Other T-lymphocytes kill any type of host cells that are
harboring viruses and other microbes in the cytoplasm.
•Cellular immunity is mainly intracellular.
Non – specific / Innate Host Defence against Viruses
The Main Host defences in viral infections are
1. Interferons 2. Mx Proteins and 3.Complements and
4. Natural Killer cells (NK cells)
Interferons
 Anti-viral proteins produced by virus-infected cells
(eventually died)
 Alert system to prevent virus from infecting other
cells and to stimulate certain lymphocytes
•It induces the expression of Interferon – stimulated genes
within the ―recipient‖ or ―target‖ cell by means of signal
cascade which inhibit viral replication.
•Species-specific for host cells
Anti-viral activities triggered by α- and β-interferons
include the following:
• Activation of genes that encode antiviral proteins,
such as dsRNA-dependent protein kinase R and
RNase L.
• Stimulation of production of major histocompatibility
(MHC) class I molecules and proteasome proteins;
these molecules enhance the presentation of viral peptides on
the infected cell surface to T cells.
• Activation of NK cells
• Induction of apoptosis

Gamma- (γ-) interferon,is produced mainly by T cells and NK

cells when triggered by certain molecules (e.g. interleukin-2)
released during immune responses.
The complement pathway can be
activated by either of three different
pathways:
1. Classical pathway (specific immune
system)
2. Lectin Pathway
3. Alternative (non-specific immune
system)
Natural killer (NK) cells
NK cells are present throughout the body, but mainly
in the blood.
They recognize changes in the surface molecules of virus-
infected cells as a result of infection, though they do not
recognize specific antigens, unlike B cells and T cells.
After recognizing virus-infected cells as target cells, NK
cells are able to bind to them and kill them.
NK cells kill their target cells either by releasing
perforins, which are proteins that are inserted into the
plasma membrane of the virus-infected cell, or by
inducing apoptosis. Also, on binding to infected cells, NK
cells release γ-interferon.
 VIRAL EVATION OF HOST DEFENSE
Viruses might be very tiny and have no brains, but when it
comes to survival games no one can call them dumb.
1. Mutation of T- cell target
•Some viruses, such as HIV, are too smart for our immune
system and learned a trick or two to fool our T-cells.
• HIV constantly changes (mutates) the regions of its
proteins that are normally targeted by T-cells and antibodies.
•So the body immunity acquired against such a disease lasts
only until the next mutation of the virus. The influenza virus
is similar to HIV in that regard, but luckily not as lethal.
• Making an effective vaccine against these viruses is
therefore very difficult.
2. Inhibition of Interferons
Epstein-Barr virus (the virus responsible for
glandular fever), produce molecules to inhibit the
action of interferons (the anti-viral cytokines)

3. Interfering with antigen presentation

Another strategy involves interfering with the
process of antigen presentation, so the T-cells
never get a chance to meet and identify the virus
and stay unaware that the body is infected.
4. Flagging protein complex
•Some viruses use to produce their own
version of the flagging (marking) protein
complex that is presented to the T-cells.
•Unlike the human protein, this viral
version does not bind to T-cells
lymphocytes.
•This way the process of presentation
completely fails and the viruses evade
detection by the lymphocytes.
5. Viral Cytokines production
•Some can even produce their own
cytokines so they compete with the human
ones and confuse the immune response.
•They can also produce receptors for
human cytokines to remove them quickly
from the infection site and silence signal,
so defence cells are not attracted to
infected area.
6. Evasion of Compliments
The strategies adopted by some viruses to evade compliments
action include:
a. Targeting the recognition molecules for the avoidance of
detection (The complement system is primarily activated by
the recognition of pathogens by the pattern recognition
molecules of this system),
Recognition molecules are: antibodies (IgM, IgG3, and
IgG1), C-reactive protein (CRP), serum amyloid P
(SAP) or SIGN-R1 (a C-type lectin)
b. Targeting key enzymes and complexes of the complement
pathways like C3 convertases and C5b-9 formation – either by
encoding complement regulators or by recruiting membrane-
bound and soluble host complement regulators,
c. Cleaving complement proteins by encoding protease, and
inhibiting the synthesis of complement proteins.