VIRUS - HOST

INTERACTIONS
Dr.T.V.Rao MD
Professor of Microbiology

Dr.T.V.Rao MD

Agents That Cause Disease.

Pathogens

Viruses

Bacteria

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Parasites

Fungi

The Immune Response to

Infectious Disease

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Important General Features of Immunity to

Pathogens.
Defense against pathogens is mediated by both
innate and specific immunity.
The innate immune response to pathogens plays an
important role in determining the nature of the specific
immune response.
The immune response is capable of responding in
distinct and specialized ways to different pathogens in
order to combat these infectious agents most
effectively.
Dr.T.V.Rao MD

Viruses
Obligatory intercellular
pathogens that replicate
within cells.
Use the nucleic acid and
protein synthetic
machineries of the host
cell.
Infect a variety of cell
populations by utilizing
normal cell surface
molecules as receptors to
enter cell.
Dr.T.V.Rao MD

Virus - Host
Virus effect Host can cause
No effect
Cell damage or Death

Polio cell death paralysis

proliferation in Moll scum
contagiosum
Malignant transformation oncogenic
virus
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Humans react to Viral Infections

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Newton's

rd
3

Law works

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Dr.T.V.Rao MD

Non Immunological
responses

Phagocytosis
Body Temperature.
Hormones
Malnutrition
Age Young Old
are more prone for
viral Infections
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BARRIERS TO INFECTION
Inherent Barriers
The host has a number of barriers to infection that are
inherent to the organism. These represent the first line of
defense which function to prevent or limit infection.
Skin
The skin acts a formidable barrier to most viruses and
only after this barrier is breached will viruses be able to
infect the host.
Lack of Membrane Receptors
Viruses gain entry into host cells by first binding to
specific receptors on cells
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Mucus
The mucus covering an epithelium acts as a
barrier to prevent infection of host cells. In some
instances the mucus simply acts as a barrier but
in other cases the mucus can prevent infection
by competing with virus receptors on cells. For
example, orthomyxo- and paramyxovirus
families infect the host cells by binding to sialic
acid receptors. Sialic acid-containing
glycoproteins in mucus can thus compete with
the cell receptors and diminish or prevent
binding of virus to the cells.
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Ciliated epithelium
The ciliated epithelium which drives the
mucociliary elevator can help diminish
infectivity of certain viruses. This system
has been shown to be important in
respiratory infections since, when the
activity of this system is inhibited by drugs
or infection, there is an increased infection
rate with a given inoculum of virus.
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Low pH
Low pH
The low pH of gastric
secretions inactivate
most viruses.
However,
enteroviruses are
resistant to gastric
secretions and thus
can survive and
replicate in the gut.
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The Immune System

The principal function of the immune
system is to protect the host against
pathogenic microbes.
Immunity may be innate or specific.

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Components of Human Immune

System

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Virus can cause Chromosomal

Damage
Chromosomal
Injury damage
Measles Mumps,
Adenovirus,CMV
Varicella virus
Damage to
chromosomes of
the Host C 17
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Pathogens & Disease

Pathogens are defined as microbes
capable of causing host damage.
When host damage reaches a certain
threshold, it can manifest itself as a
disease.
The evolution of an infectious disease in an
individual involves complex interactions
between the pathogen and the host.
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Evasion of Immune
Mechanisms by Viruses
Viruses can also
escape immune
attack by changing
their antigens.
A large number of
viruses evade the
immune response by
causing generalized
immunosuppression.
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Inclusion Bodies
Inclusion bodies are
nuclear or cytoplasmic
aggregates of stainable
substances, usually
proteins. They typically
represent sites of viral
multiplication in a
bacterium or a eukaryotic
cell and usually consist of
viral capsid proteins.

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Inclusion Bodies
( Elementary Bodies )
Inclusion bodies differ
size, shape, Location,
staining ,properties,
Some are seen under
microscope
In Cytoplasm Pox virus
Nucleus is affected
Herpes virus

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Negril Bodies

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Staining the Virus in Inclusion

bodies
Giemsa staining can
produce Acidophilic /
Basophilic inclusions
Esinophilic inclusions
Negril bodies in brain
cells in Rabies
Mollusca Bodies
moll scum
contagiosum
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Pathogenesis of Viral Infection

In apparent ( Sub clinical )
Apparent
( Clinical )
Acute
Sub acute
Chronic
Can produce latency
Herpes zoster virus in nerve toot ganglion,
Kuru Human slow virus infection.
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How Virus enter the Host

Through
Respiratory tract
Gastro Intestinal
tract
Skin,
Conjunctivae
By sex contact
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Respiratory tract
Small pox
Chicken pox
Influenza
Rhinovirus
Rhinovirus

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Gastro Intestinal tract

Enterovirus,
Adenovirus
Reovirus
Hepatitis A E
Rota virus
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Skin
Papilloma virus
Cow pox
Molloscom
contagiosum
Animal Bite Rabies
Injections Hepatitis
B infections.
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Most common Viral Infections

Arbovirus
Mosquito
bite
Dengue
Chikungunya
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Other Routes of Entry

Conjunctiva
Adenovirus,
Genital tract
sexual contact
HIV
Congenital
Infection
Rubella and CMV

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Spread of Virus
Spread from various
sources
Lymph nodes, Blood
stream, Reach target
organs.
Viremia locate to
various organs.

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Incubation period
May be short,
long
Variable.
Depend on site of entry and
site of lesions.
Common cold very rapid
onset.
Chicken pox and
Poliomyelitis 10 20
days
Arbovirus 5 6 days
Hepatitis B 2 6 months
AIDS ?
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Slow Virus many years

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How Host Responds

Non Specific
Immunity
Humoral
Cell
mediated.
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Specific Immune Response to Viruses

Mediated by a combination of humoral and cell
mediated immune mechanisms.
Humoral mediated immune response.
Antibodies specific for viral surface antigens are

often crucial in containing the spread of a virus

during acute infection and in protecting against reinfection.
Specific antibodies are important in defense
against viruses early in the course of infection and
in defense against cytopathic viruses that are
liberated from lysed infected cells.
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Specific Immune Response to

Viruses
Cell-mediated
immune responses.
Most important in host
defense, once a viral
infection is established.
CD8+ Tc cells
(Cytotoxic T
lymphocytes; CTLs)
and CD4+ th1 cells
(helper T lymphocytes)
are the main
components of cell
mediated antiviral
defense.
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CD8+ T and CD4+ T

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Humoral Immunity
Immunoglobulin Ig G
Ig M Ig A

Ig A Mucosal
surface secretary
immunoglobulin
Antibodies
neutralize with
help of
complement.
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HIV destroys CD 4 cells

Cell Mediated
Immunity
Delayed
Hypersensitivity

HIV destroys
CD 4
Lymphocytes.
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Evasion of Immune
Mechanisms by Viruses
Viruses have evolved
numerous
mechanisms for
evading host
immunity.
A number of viruses
have strategies to
evade complementmediated destruction.
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Evasion of Immune
Mechanisms by Viruses
Viruses can also
escape immune
attack by changing
their antigens.
A large number of
viruses evade the
immune response by
causing generalized
immunosuppression.
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HUMORAL COMPONENTS INVOLVED IN

RESISTANCE TO VIRAL INFECTIONS
Nonspecific
A number of humoral components of the nonspecific
immune system function in resistance to viral infection.
Some of theses are constitutively present while others
are induced by infection.
Interferon (IFN)
IFN was discovered over 40 years ago by Issacs and
Lindemann who showed that supernatant fractions from
virus-infected cells contained a protein that could confer
resistance to infection to other cells. This substance did
not act directly on the virus, rather it acted on the cells to
make them resistant to infection (Figure 1).
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Immunity Notsustanbale in Influenza Infection

Need Vaccination to New strains

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Types of Interferon's

Alpha
Beta
Gamma

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Interferon's
There are three types of interferon, IFNalpha (also known as leukocyte
interferon), IFN-beta (also known as
fibroblast interferon) and IFN-gamma (also
known as immune interferon). IFN-alpha
and IFN-beta are also referred to as Type I
interferon and IFN-gamma as Type II.
There are approximately 20 subtypes of
IFN-alpha but only one IFN-beta and IFNgamma. Molecular
wt. MD17,000
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Interferon's
Isaacs Linder Mann

Interferon protection against viral

infection Contain Host coded proteins
Produced by viral and Non viral
inducer On exposure to Interferon
cell produce translation inhibiting
proteins TIP Inhibits translation of m
RNA But no effect on cell mRNA Viral
transcription inhibited .
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Biological effects of
Interferon's
Antiviral effects.
Antimicrobial effects.
Cellular effect ts Inhibition of cell growth
and proliferation. and of DNA and protein
synthesis Expansion MHC antigens
Immunoregulatory effects Enhanced
cytotoxic activity of NK, K and T
Suppression of DTH.
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Interferon's
Hum Interferon alpha leukocyte interferon
Produced by leukocytes 16 subspecies
Beta Interferon Inf Produced by Fibroblast and
epithelial cells.
Gamma Interferon Inf produce by
lymphocytes,
Produce Immuno modulation and ant
proliferative function
Resist heating at 56 - 600 c for 30 6o mt
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Interferon's
Interferon's are species specific
RNA viral infections are better inducers
than DNA virus
Potent are
Toga viridae Vesicular stomatitis virus
and Sendai virus.
Production starts in > 1 hour and increases
in 6 12 hours.
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Uses of Interferon's
Molecular wt. 17,000
Used in Prophylaxis
and treatment
Non toxic and Non
antigenic
Anti cancer agent
lymphomas
Used in Hepatitis B
and C infections.
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Complement
Most viruses do not fix complement by the
alternative route. However, the interaction
of a complement-fixing antibody with a
virus infected cell or with an enveloped
virus can result in the lysis of the cell or
virus. Thus, by interfacing with the specific
immune system, complement also plays a
role in resistance to viral infections.
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Cytokines
Cytokines other than IFN also may play a
role in resistance to virus infection. Tumor
necrosis factor alpha (TNF-), interleukin1 (IL-1) and IL-6 have been shown to have
antiviral activities in vitro. These cytokines
are produced by activated macrophages
but their contribution to resistance in vivo
has not been fully elucidated.
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Specific
Antibody produce by the specific immune system is
involved primarily in the recovery from viral infection and
in resistance to subsequent challenge with the virus.
IgG, IgM and IgA antibodies can all play a role in
immunity to virus infection but the relative contributions
of the different classes depends on the virus and the
portal of entry. For example, IgA will be more important
in viruses that infect the mucosa while IgG antibodies will
be more important in infections in which viremia is a
prominent feature. Antibodies can have both beneficial
and harmful effects for the host.
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Laboratory Diagnosis of
Viral Diseases
Different from Bacterial Infections

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Virological Tests
An Overview

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Why Difficult
In the past Growth of Virus was not rapid.
Diagnosis becomes routine today due to
availability of Rapid method.
Important in HBV and HIV infections.
Rubella in pregnant women.
Simple methods Microscopy and
detection of inclusion bodies.
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Diagnostic Methods in
Virology
1. Direct Examination

2. Indirect
Examination (Virus
Isolation)
3. Serology
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Microscopy
Light Microscopy
elementary bodies
Electron Microscopy
Rota viral detection
Florescent
Microscopy Direct /
Indirect

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Direct Examination
1. Antigen Detection

immunofluorescence, ELISA etc.

2. Electron Microscopy

morphology of virus particles

immune electron microscopy

3. Light Microscopy

histological appearance
inclusion bodies

4. Viral Genome Detection

hybridization with specific

nucleic acid probes
polymerase chain reaction (PCR)
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Indirect Examination
1. Cell Culture

cytopathic effect (CPE)

haemabsorption
immunofluorescence

2. Eggs

pocks on CAM
haemagglutination
inclusion bodies

3. Animals

disease or death

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Serology
Detection of rising titers of antibody between acute and
convalescent stages of infection, or the detection of IgM in
primary infection.
Classical Techniques

Newer Techniques

1.
2.
3.
4.
5.

1.
2.
3.
4.
5.

Complement fixation tests (CFT)

Haemagglutination inhibition tests
Immunofluorescence techniques (IF)
Neutralization tests
Counter-immunoelectrophoresis

Radioimmunoassay (RIA)
Enzyme linked immunosorbent assay (EIA)
Particle agglutination
Western Blot (WB)
RIBA, Line immunoassay

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Virus Isolation
Cell Cultures are most widely used for virus isolation, there are 3
types of cell cultures:
1. Primary cells - Monkey Kidney
2. Semi-continuous cells - Human embryonic kidney and skin
fibroblasts
3. Continuous cells - HeLa, Vero, Hep2, LLC-MK2, MDCK
Primary cell culture are widely acknowledged as the best cell culture
systems available since they support the widest range of viruses. However,
they are very expensive and it is often difficult to obtain a reliable supply.
Continuous cells are the most easy to handle but the range of viruses
supported is often limited.
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Cell Cultures
Growing virus may produce
1. Cytopathic Effect (CPE) - such as the ballooning of cells or
syncytia formation, may be specific or non-specific.
2. Haemabsorption - cells acquire the ability to stick to
mammalian red blood cells.
Confirmation of the identity of the virus may be carried out using
neutralization, haemabsorption-inhibition or immunofluorescence
tests.

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Detection of Viral antigen

Use of
Immunofluorescence
Imunoelectrphoresis
Radio immunoassay
RIA
ELISA
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Serology
Since the isolation and identification of viruses is
not commonly done in the clinical laboratory, the
clinical picture and serology plays a greater role
in the diagnosis of viral disease. The major types
of antibodies that are assayed for are
neutralizing, haemagglutination inhibiting and
complement fixing antibodies. Complement
fixing antibodies follow the kinetics of IgM and
are most useful in indicating a current or recent
infection
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Serology
The development of
antibodies to different
components of the
virus is used in
staging the disease.
For example in
hepatitis B and HIV
infections this
approach is used.
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Serological Diagnosis
Detection of
Immunologlublins Ig
G. Ig M Ig A
Raise of titers Ist
sample later sample
(convalescent
sample) tested after
10 14 days Raise of
titer is diagnostic
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ELISA for HIV antibody

Microplate ELISA for HIV antibody: coloured wells indicate reactivity

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Western Blot
HIV-1 Western Blot

Lane1: Positive Control

Lane 2: Negative Control
Sample A: Negative
Sample B: Indeterminate
Sample C: Positive

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Polymerase Chain Reaction

Advantages of PCR:
Extremely high sensitivity, may detect down to one viral genome
per sample volume
Easy to set up
Fast turnaround time

Disadvantages of PCR
Extremely liable to contamination
High degree of operator skill required
Not easy to set up a quantitative assay.

A positive result may be difficult to interpret, especially with latent

viruses such as CMV, where any seropositive person will have virus
present in their blood irrespective whether they have disease or not.
.

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Schematic of PCR

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Each cycle doubles the copy number of the target

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Growing Technologies
Molecular
methods
Probes
Polymerase
chain reaction
Can produce
Rapid Highly
scientific Specific
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Regular Methods in Use

Egg inoculation
Pox virus,
Influenza
Into tissue
culture
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Advantages of Molecular
Methods
Increases
Sensitivity
and
Specificity.
PCR
RT PCR

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Antiviral Agents
Restricted spectrum
No standardized in-vitro susceptibility tests
Most inhibit replication. Cure depends on host
immune system to eradicate. If patients are
immunocompromized, may have recurrences.
Many need to be activated by viral and cellular
enzymes before exerting antiviral effect. Activity
of enzymes and concentration of substrates will
influence the efficacy.

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Nucleoside Analogues
General Mechanism of Action
1. Taken up by cells
2. Converted by viral and cellular enzymes to the
triphosphate form
3. The triphosphate form inhibits:
1. DNA polymerase
2. Reverse transcriptase
3. RNA polymerase

4. Or it may get incorporated into growing DNA

leading to abnormal proteins or breakage.

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Ganciclovir

Mechanism like Acyclovir

Active against all Herpes viruses including CMV
Low oral bioavailability given I.V.
Most common adverse effect: bone marrow
suppression (leukopenia 40%,
thrombocytopenia (20%) and CNS effects
(headache, behavioral, psychosis, coma,
convulsions).
1/3 of patients have to stop because of adverse
effects
Drug of choice for CMV infections: retinitis,
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pneumonia, colitisDr.T.V.Rao MD

Anti-retroviral Agents
Zidovudine (AZT)
Cellular enzyme phosphorylate to the triphosphate form
which inhibits RT and causes chain termination
Adverse effect:
Granulocytopenia and anemia: 45% in AIDS but 5% if
asymptomatic HIV
Severe headache, nausea, insomnia, myalgia
mortality & opportunistic infections, gain weight, better
quality of life, delays signs and symptoms of AIDS

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Protease Inhibitors
Produce non-infectious particles or virions
Reduces the number of new rounds of infection
in susceptible cells
To be effective must be prolonged, profound and
constant.
Pharmacokinetics important to maintain constant
concentrations within the effective range.
Metabolic adverse effects (DM, hyperglycemia)
and GI (diarrhea, pain vomiting).

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Drug Examples
Tamiflu Recently sold to 40
countries to battle avian flu
Prevents the mature viruses
from leaving the cell
It is a neuraminidase
inhibitor, it works on both
influenza A and B
Neuraminidase is an
enzyme found on the virus
which cleaves sialic acid
from cell membrane, leading
to a more effective release
of viruses. mechanism
Dr.T.V.Rao MD

Other Drugs
Amantadine

Interferon's

Prevents uncoating (?)

Antiviral, anticancer and
&/or assembly
immunomodulating
CNS Toxicity due to
Several sites of action in
dopaminergic action
viral cycle but mainly
Prophylaxis of Influenza A
inhibit translation of viral
during epidemics.
proteins
If used within 48 hours
Toxicity: flu-like
may help cure Influenza
syndrome, BM
infection
suppression; CNS
Rimantadine: analog with
less CNS toxicity
Hepatitis B and C
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Created by Dr.T.V.Rao MD
for Basic Learning of
Medical Virology
email
doctortvrao@gmail

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