HOST DEFENSE AGAINST VIRAL INFECTION-

ANIMALS
• Primary defenses- physical and chemical barriers
-skin
-mucous secretions
-tears
-acid pH
-surface cleansing mechanisms (swallowing,
blinking)

   
IMMUNE DEFENSES
• Three critical processes in the immune defense:
-recognition
-amplification
-control
• The immune response to viral infection consists of:
Innate (nonspecific) defense: first line of immune
defense, responds to any infection, recognizes
characteristics common to microbial invaders,consists
of interferons, complement, natural killer cells, dictates
the adaptive response
Adaptive (specific) defense: antibody response and the
lymphocyte-mediated response also called the humoral
and
  cell-mediated responses 
Innate and adaptive
immune responses:

   
 The innate immune response:
• Can be activated rapidly and functions within hours
of a viral infection.
• Continued activity is damaging to the host.
• Considerable interplay occurs between the adaptive
and innate immune defenses.
Important components are:
-cytokines
-complement
-collectins
-natural killer (NK) cells

   
 The adaptive immune response:

• Differentiates self from nonself, tailored to the

particular invader
• Has ‘memory’; subsequent infection by the same
agent are met with a robust and highly specific
response that stops the infection
• Consists of the:
antibody response - humoral response
lymphocyte mediated response- cell-mediated
response

   
   
O’Neill, Scientific American, Jan 2005
pp. 38-45
 The inflammatory response:

• Essential in initiating immune defenses

• Cell and tissue damage caused by infection

induces the inflammatory response

• Provides communication with the components of

the immune system

• Characterized by redness, heat, swelling and pain

   
The inflammatory assault is initiated by Toll-like
receptors

   
Inflammation can be initiated in
several ways:
• By interferon released by
immature dendritic cells
• Locally produced cytokines,
such as interleukin-1, tumor
necrosis factor- α and
interferon-γ control the
reactions that occur during
inflammation
• Inflammatory cytokines also
activate B and T cells that are
needed for the adaptive
response
   
Cytokines:
• Regulatory proteins that mediate intercellular
communication during an antiviral defense.
• Their presence is one of the first indicators that the
host has been infected.
• They act locally, near the cells that make them.
• They control inflammation, induce and antiviral state
in cells and regulate the adaptive immune response.
• They exert their activities by binding to specific
receptors and activating gene expression.
• Three types of interferons are the most important
cytokines in the innate response to viral infection.

   
 Interferons

Ifn-γ is induced only when certain lymphocytes are

stimulated to replicate and divide after binding a
foreign antigen
Ifn-α and Ifn-β are induced by viral infection of any cell
type
   
 Interferons
• Ifn is induced by accumulation of double stranded
RNA (dsRNA).
• Ifn induces gene expression at the transcriptional
level after binding to specific cell surface receptors.
• A cell that is bound to interferon and responds to it
is in an antiviral state.
• Ifn induces expression of more that 100 genes,
products of many of these genes possess broad
spectrum antiviral activity.
• They lead to cell death by apoptosis or programmed
cells death, limiting cell to cell spread of virus.
• Production of large amounts if Ifn causes common
symptoms such as fever, chills, nausea, etc.
   
Interferon induced antiviral responses:
• Both viral and cellular protein synthesis stops in Ifn
treated cells.
• This is dues to two cellular proteins, ds-RNA activated
protein kinase (Pkr) and ribonuclease L (RNase L).
• Pkr is a serine/threonine kinase that has antiviral
properties, as well as antiproliferative and antitumor
functions.
• Activated Pkr phosphorylates the alpha subunit of the
translation initiation factor eIF2, inhibiting translation.
• RNase L is a nuclease that can degrade cellular and viral
RNA; its concentration increases after Ifn treatment.

   
 Interferon induced antiviral responses:
• RNase L concentration increases 10-1,000 fold after
Ifn treatment, but is inactive unless 2’-5’-oligo(A)
synthetase is produced.
• 2’-5’-oligo(A) synthetase produces 2’, 5’ oligomers of
adenylic acid, only when activated by dsRNA.
• These poly(A) oligomers then activate RNase L,
which degrades all host and viral mRNA in the cell.
• RNase L participates not only in Ifn-mediated
antiviral defense, but also in apoptosis.
• Ifn is a broad spectrum, highly effective antiviral
agent. However, viruses have developed numerous
mechanisms for inhibiting interferon action.
   
   
 The adaptive immune response:
• Humoral response
Consists of lymphocytes of the B-cell lineage
Interaction of a specific receptor on precursor B
lymphocytes with antigens promotes differentiation
into antibody secreting cells (plasma cells).
• Cell-mediated response
Consists of lymphocytes of the T-cell lineage
Cytotoxic T cells (Tc cells) and T-helper cells (Th
cells) are the key effectors of this response.

   
 Cell-mediated response cont.

• T lymphocytes recognize antigens on the surface of

self cells.
• The antigens on self cells can be recognized only by a
receptor on the surface of T cells when they are bound
to the MHC family of membrane proteins.
• The Th cells recognize antigens bound to MHC class II
molecules and produce powerful cytokines that affect
other lymphocytes (B and T cells) by promoting or
inhibiting cell division and gene expression.
• Once activated by Th cells, Tc cells differentiate into
CTLs that can kill virus infected cells.

   
   
The antigen receptors on the surface of B and T cells

B cells have about 100,000

molecules of a single antibody
receptor per cell, which has
specificity for one antigen
epitope.

T cells bearing the surface

membrane protein CD4 always
recognize peptides bound to
MHC class II proteins and
function as Th cells.

T cells bearing the surface

membrane protein CD8 always
recognize
  peptide antigens
 
bound to MHC class I proteins
 Endogenous antigen processing: MHC class I peptide
presentation
• Intracellular proteins of host and virus are
marked for
degradation by ubiquitination and are degraded
by the
Proteasome.

• The resulting viral peptides are transported

into the ER
lumen by the Tap1-Tap2 heterodimeric
transporter.

• In the ER lumen, viral peptides associate with

newly
synthesized MHC class I molecules.
   
• MHC class I-peptide complex is transported to
Endogenous antigen processing: MHC class I peptide presentation

   
 Exogenous antigen processing: MHC class II peptide
presentation
• MHC class II complex is prevented form
binding to viral peptides in the ER by
association with the invariant chain.
• The complex is transported through golgi
where the invariant chain is removed, activating
the MHC class II complex.
• The peptides are derived from extracellular
proteins that enter the cell by endocytosis.
• Viral proteins are degraded in the lysosomes
by proteases that are activated by low pH.
• Endosomes fuse with vesicles containing MHC
class II.
•   On the surface of the   cell the MHC class II
 Exogenous antigen processing: MHC class II peptide
presentation

   
ANTIVIRAL DRUGS

•Because viruses are obligate intracellular parasites,

identification of safe and effective antiviral therapies is
difficult.

•The best antiviral drugs inhibit a specific step in viral

replication or pathogenesis.

•Drug discovery can be accomplished by screening or

rational design.

•The emergence of virus mutants resistant to antiviral

drugs is a serious problem.

•Combination of targeted delivery strategies to control

toxicities
 
and resistance.  
   
 The pathway for drug discovery

   
Antiviral
compounds
Many well-known antiviral
compounds are
nucleoside and nucleotide
analogs:
• Acyclovir is a nucleoside
analog similar to
guanosine, but contains
an acyclic sugar group
• AZT is a nucleoside
analog similar to
thymidine, but contains an
azide group
   
Acyclovir:
•Close to a perfect antiviral drug (specific, nontoxic).
•Highly effective against herpes simplex virus (HSV),
less so against varicella-zoster virus (VZV).
•Higly selective and extremely safe.
•Acyclic guanine derivative that inhibits viral DNA
synthesis.
•It is a prodrug, a precursor of the antiviral
compound.
•Activation of the drug requires three kinase
activities to be present in the cell to convert
acyclovir to a triphosphate derivative, the actual
antiviral
  drug.  
Activation of acyclovir:
• Acyclovir has no effect
on host DNA replication
because the first kinase
activity is not found in an
noninfected cell.
• Cellular enzymes
complete the synthesis.
• Incorporated into viral
DNA, lacks the 3’ OH of
the sugar ring.
• Growing DNA chain
terminates, replication is
blocked.
   
Chemical structures of effective antiviral compounds:

   
Commercially available antiviral drugs:

   
Steps in replication of HIV targeted by antiviral drugs:

   
Antivirals against HIV:
• Azidothymidine (AZT)
Dideoxy analog of thymidine
Inhibits viral DNA synthesis
• Efficiently phosphorylated
to triphosphate by cellular
kinases
• AZT monophosphate competes with thymidine
monophosphate
• Much less selective than acylovir and has side effects
Does not eliminate previously incorporated provirus
   
 Protease inhibitors
•Uncleavable mimics of gag-pol polyprotein
•Inhibits HIV protease
•Does not eliminate previously incorporated provirus
but does prevent further spread
•Resistance due to protease alterations

   
 Prevention & Treatment of Influenza:
Beyond Chicken Soup  
Zanamivir (Relinza): Blocks
active site of all NA proteins.
Must be inhaled as a spray,
but prevents flu symptoms if
given before infection
& reduces symptoms if given
shortly after infection. Has no
side effects.
Other Anti NA Drugs in
development can be taken as
pills and appear to be very
effective in reducing illness if
taken early in infection. These
drugs look very promising in
clinical trials and have no
adverse effects.
Neuraminidase Inhibitors:
Zanamivir & Oseltamivir
• Mechanism: blocking of the active site of
neuraminidase; prevents removaal of
sialic acid residues and results in
clumping of viral progeny
• Effective against influenza A & B.
• Effective when flu symptoms are < 2 days
old.
• Inhibitors reduce disease syndrome by 1
day.
• May decrease influenza secondary
complications
• Antiviral resistance can occur, but much
  less frequently than  with the ion channel
 Influenza Treatment with Ion
Channel Blockers Amantadine &
Rimantadine
• Effective against influenza A but not B.
• Mechanism: inhibit ion channels & viral
uncoating.
• Can reduce severity & duration of illness if
started within 48 hrs of onset of
symptoms.
• Treated persons may shed resistant virus
after 5-7 days of treatment (sometimes as
early as 2-3 days).
• Treatment should be discontinued after 3-
5 days of treatment or within 24-48 hrs
after
  disappearance  of signs/symptoms).
 Antiviral Agents for
Influenza
• Neuraminidase inhibitors appear to have
similar efficacy to the amantidine &
rimantidine ion channel blockers for
prevention & treatment of influenza
• Neuraminidase inhibitors have Less
Central Nerveous System side effects,
but more Gastro-Intestinal effects
• Neuraminidase Inhibitors are more
expensive, but there is less risk of
inducing virus resistance.
Very rapidly growing field of research.