Medical virology

By Rebie K

Email;- rebiekedir11@gmail.com
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 Chapter I
Introduction to Virus
Learning objectives
Upon completion of this chapter, the student will be able to:

1. Define virus.

2. Describe structure & properties of medically important viruses

3. Illustrate classification of medically important viruses

4. Explain virus infectious cycle

5. Describe antiviral agents and their mechanism of action

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 1.1. Introduction to Viruses
Discovery of viruses and brief history
• Concepts of viruses - less than 100 years
• Nature began to be understood , less than 50
years
• Diverse, represent a definite entity with shared
properties and concepts
• Every species of life carries viruses
• Many are harmful , some symbiotic and give
advantages to the host (drug resistance, gene
transfer, virulence factor)

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Poliomyelitis Through the Ages

1200 BC 1968
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Reaktivierung von Virusinfektionen

HSV VZV

VZV

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Viruserkrankungen

Masern
Mumps
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 Discovery of viruses…
• As tools for understanding the cellular and
molecular biology of the host organism
− Chemical nature of genes, transcription control, DNA
repair, translation of RNA, RNA processing, protein
modification, protein-protein interaction
− Cancer
− Function of cells

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 Discovery of viruses…
• Concept of Vaccination
− 1780’s E. Jenner (Cow pox vaccine)
− 1880’s L. Pasteur ( Rabies Vaccine)
− 1892 Tobacco mosaic virus
− 1898 Foot & Mouth disease virus
− 1900 Yellow fever virus
− 1916-17 Discovery of bacteriophage
− 1940 The replication of cycle of bacteriophage

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 Discovery of viruses…
• Bacteriophages and the beginning molecular
virology
• Viruses in human history
− Rabies
− Polio 1500 B.C.
− Small pox 650 B.C.
− Rabies
− Newly emerging viruses

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 Features of Viruses

Definition: Viruses are sub-microscopic, intracellular parasites

with only one type of nucleic acid

· Viral genome either DNA or RNA

· Viruses exploit host cell machinery for protein

synthesis and cellular enzymes for energy

· Viruses redirect host cell metabolism to primarily

synthesise viral proteins

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 VIRUS
• Viruses consist of a nucleic acid surrounded by
one or more proteins.
• Some viruses also have an outer-membrane
envelope. Viruses are obligate
• Intracellular parasites: they can replicate only
within cells since →
their nucleic acids do not encode many
enzymes necessary for replication or energy
production

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 Virus like Infectious particles
• Virusoids are nucleic acids that depend on helper viruses
to package their nucleic acids into virus-like particles.
• Viroids are small naked ss RNA molecules
restricted to plants, spread from cell to cell. RNA molecule
contains no protein encoding genes. So totally dependent on
host cell functions for replication
• II. Prions are abnormal infectious protein molecules that
can spread and change the structure of their normal
counterparts (cellular proteins) →transmissibility

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 1. Introduction to virus

1.1 Definition and properties of a virus

• Viruses are filterable agents
• Viruses are the smallest infectious agents

(Size: 20 – 400 nm in diameter)

• Are obligatory intracellular parasites

• Cannot make energy or proteins independent of a host cell

• Viral genomes may be RNA or DNA , but not both

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 Nature of Viruses
• Infectious agent and obligate intracellular parasite
• Have infectious cycle
• Able to be transmitted: Transmissible
• Able to redirect genetic and metabolism apparatus
• Genome either DNA or RNA
• Have major features of Cellular organisms:
• life cycle, defined stage of development,
• organization, genetic variation
• Do not possess some features:
• No machinery of metabolisms and protein synthesis
• Can not reproduce outside the host cell

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 Nature of Viruses…
• Viruses differ from toxins
• Toxins cause disease, do not multiply, do not
increase in quantity as a biological disease
• Differ from plasmids: Self replicate but lack
protective structures
• Differ from other intracellular parasites:
infection maintained by cell and does not have
eclipse period

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 Introduction…

• Viral components are assembled & do not replicate by

division
• Viruses are not living
• Virus must be infectious to endure in nature

1.2 Evolutionary origin of viruses

• How did these become independent genetic entities?

► Not clearly well elucidated, however three hypothesis:

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 Introduction…

A. Regressive theory

• Viruses are degenerate forms of intracellular parasites

• E.g. Leprosy bacillus, rickettsiae & chlamydia have all

evolved in this direction

• Lack their own rRNAs or protein synthesis machinery

Controversy on how RNA virus evolved???

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B. Progressive theory

• From cellular RNA and DNA components

• Normal cellular nucleic acids that gained the ability to

replicate autonomously & hence to evolve

• DNA viruses came from plasmids or transposable elements

►They then evolved coat proteins & transmissibility

• Retroviruses derived from retro-transposons & RNA virus

from mRNA
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C. Co - evolution theory

• Viruses coevolved with life ( free living cells)

• All of the above theories could be could be correct!

• No compelling reason to think that RNA viruses have

evolved in the same way as DNA viruses.

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STRUCTURE OF HIV

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Structural Components of Viruses

lipid membrane (envelope)

envelope proteins

nucleic acid

Protein shell (nucleocapid)

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Definitions:
Virion - physical particle of the virus
Core - nucleic acid and tightly associated proteins within the virion VIRUS
Capsid - protein shell around NA or core STRUCTURE
Capsomere - protein subunit making up the capsid
Nucleocapsid - core and capsid
Envelope - lipid membrane found on some viruses,
often derived by budding from infected cells.
Peplomer - ("spike”)- morphological unit projecting
from the envelope or surface of a naked virion

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 VIRUS STRUCTURE
• The capsid (coat) protein is the basic unit of structure;
functions that may be fulfilled by the capsid protein are to:
– Protect viral nucleic acid
– Interact specifically with the viral nucleic acid for packaging
– Interact with vector for specific transmission
– Interact with host receptors for entry to cell
– Allow for release of nucleic acid upon entry into new cell
– Assist in processes of viral and/or host gene regulation

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1.2 Definitions of terminologies

• Capsid - Protein coat that enclose the NA genome

• Nucleo - capsid - Capsid plus NA

• Structural units - Basic protein building blocks of the
coat

• Capsomers :
 are viral morphological units

 Seen in electron microscope on the surface of

Icosahedral viral particles
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  Represent clusters of polypeptides

 Their number vary from one virus group to the other

• Envelope

 lipid containing membrane that surrounds some viral

particles

 Acquired during viral maturation by a budding process

 Virus encoded glycoprotein are exposed on the surface

of the envelop
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• Virion
 Complete and infective viral particle
 Transfers viral NA from cell to cell
• Defective virus

Viral particle functionally deficient in some aspects of

replication E.g. HDV requires helps from HBV
• Prions are abnormal infectious protein molecules that
can spread & change the structure of their normal
counterparts (cellular proteins) →transmissibility
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1.3 Structure of medically important viruses
• Components of complete virus particles

Enveloped viruses comprise:

Capsomers, envelope, nucelo- capsid
(nucelic acid plus capsid)

Naked capsid viruses comprise:

Capsomers & nucelo- capsid
( See fig 1 &2 …)
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 Properties of naked capsid viruses

Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005.
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 Properties of enveloped viruses

From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005, Fig. Box 6-5.
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• Viral glycoproteins
Envelope contains glycoproteins
They are virus- encoded but sugars from host cell
1 Glycoproteins attach virus to host cell by interacting with a
cellular receptor
2 Involved with membrane fusion step of infection
3 Serve as important viral antigens
4 Involved with neutralizing antibodies
5 Serve specific functions like HA and NA influenza virus

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 Basic virus structure

Fig. 1. From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005,

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Fig 2. components of complete virus particle : A) Enveloped virus with icosahedral
symmetry. B) Virus with helical symmetry.
Source : Medical Microbiology, 24 th ed; 2000

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 Basic virus structure

http://www.med.sc.edu:85/mhunt/intro-vir.htm

From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc.,
2005, Fig. 6-5.
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 Structures compared

Fig. Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005

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 Viral Protein Shell

cubic complex helical

icosahedral (Variola virus) (Influenza virus)
(Herpes simplex virus)

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 Capsid Symmetry : 3 types

(1) Cubic symmetry: Isometric or spherical

− Icosahedron pattern: the most efficient
arrangement subunits in closed shell

− Advantage
- The subunits can be smaller and thus
economizing on genetic information
- Avoids physical restraints which prevents
the tight packing o subunits in other
symmetry

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 Capsid Symmetry…

2) cosahedral or Cubic Symmetry

Most efficient arrangement for subunits in a closed shell
Animal viruses in microscopy appear spherical but are
icosahedral
An icosahedron has 20 faces, 12 vertices Five fold, three
fold or twofold axes
Exactly 60 identical structural subunits

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Symmetry of virus
 Capsid Symmetry…
(2) Helical symmetry
− Rod like or thread like
− Filamentous viruses

Helical (Influenza virus)

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 Helical Symmetry--
E.g. Helical animal viruses
• The simplest way to arrange multiple, identical protein subunits is to use rotational

symmetry & to arrange the irregularly shaped proteins around the circumference of a circle

to form a disk

• Multiple disks can then be stacked on top of one another to form a cylinder, with the virus

genome coated by the protein shell or contained in the hollow centre of the cylinder

• This category includes many of the best known human pathogens, e.g. influenza virus,

mumps & measles viruses, & Rabies virus

• All helical animal viruses possess single-stranded, negative-sense RNA genomes 40

Classification of viruses

1. Type of Nucleic Acid

 DNA or RNA (Ss /Ds)

 Strategy of replication

2. Size & morphology

 Type of symmetry ( Icosahedral, Helical . Complex )

 Number of capsomers

 Presence or absence of envelope

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3. Presence of specific enzymes

E.g: - RNA and DNA polymerase

- Neuraminidase

- Reverse transcriptase

4. Host tissue or cell tropism

E.g: Hepatitis viruses, HIV, etc

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5.Mode of transmission e.g. Arboviruses
6. Host range E.g. Animal, bacteria, plant
7. Type of disease E.g. encephalitis

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 DNA VIRUSES

DOUBLE STRANDED SINGLE STRANDED COMPLEX ds

NON-ENVELOPED ENVELOPED

ENVELOPED NON-ENVELOPED PARVOVIRIDAE POXVIRIDAE

HERPESVIRIDAE
HEPADNAVIRIDAE
CIRCULAR LINEAR

PAPILLOMAVIRIDAE ADENOVIRIDAE All families shown are

POLYOMAVIRIDAE icosahedral except for
(formerly grouped together as the poxviruses
PAPOVAVIRIDAE)

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 RNA VIRUSES

SINGLE STRANDED + SINGLE STRANDED DOUBLE STRANDED

negative sense
sense

ENVELOPED NONENVELOPED ENVELOPED NONENVELOPED

ICOSAHEDRAL HELICAL ICOSAHEDRAL HELICAL

ICOSAHEDRAL

FLAVIVIRIDAE CORONAVIRIDAE PICORNAVIRIDAE ORTHOMYXOVIRIDAE REOVIRIDAE

TOGAVIRIDAE CALICIVIRIDAE PARAMYXOVIRIDAE
RETROVIRIDAE RHABDOVIRIDAE
FILOVIRIDAE
BUNYAVIRIDAE
ARENAVIRIDAE

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Positive & negative sense RNA
 Positive sense RNA can be translated directly into
protein upon uncoating of the virion in the cell
 Negative sense RNA must be transcribed by a virus
coded, virion packaged RNA dependent RNA
polymerase immediately following uncoating

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 Classification of Human Viruses
"Group" Family Genome Genome size (kb) Capsid Envelope
dsDNA
Poxviridae dsDNA, linear 130 to 375 Ovoid Yes
Herpesviridae dsDNA, linear 125 to 240 Icosahedral Yes
Adenoviridae dsDNA, linear 26 to 45 Icosahedral No
Polyomaviridae dsDNA, circular 5 Icosahedral No
Papillomaviridae dsDNA, circular 7 to 8 Icosahedral No
ssDNA
Anellovirus ssDNA circular 3 to 4 Isometric No
Parvoviradae ssDNA, linear, (- or +/-) 5 Icosahedral No
Retro
Hepadnaviridae dsDNA (partial), circular 3 to 4 Icosahedral Yes
Retroviridae ssRNA (+), diploid 7 to 13 Spherical, rod or cone shaped Yes
dsRNA
Reoviridae dsRNA, segmented 19 to 32 Icosahedral No
ssRNA (-)
Rhabdoviridae ssRNA (-) 11 to 15 Helical Yes
Filoviridae ssRNA (-) 19 Helical Yes
Paramyxoviridae ssRNA (-) 10 to 15 Helical Yes
Orthomyxoviridae ssRNA (-), segmented 10 to 13.6 Helical Yes
Bunyaviridae ssRNA (-, ambi), segmented 11 to 19 Helical Yes
Arenaviridae ssRNA (-, ambi), segmented 11 Circular, nucleosomal Yes
Deltavirus ssRNA (-) circular 2 Spherical Yes
ssRNA (+)
Picornaviridae ssRNA (+) 7 to 9 Icosahedral No
Calciviridae ssRNA (+) 7 to 8 Icosahedral No
Hepevirus ssRNA (+) 7 Icosahedral No
Astroviridae ssRNA (+) 6 to 7 Isometric No
Coronaviridae ssRNA (+) 28 to 31 Helical Yes
Flaviviridae ssRNA (+) 10 to 12 Spherical Yes
Togaviridae ssRNA (+) 11 to 12 Icosahedral Yes
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 Baltimore Classification
• The Baltimore scheme of classification distinguishes
between viruses whose genomes can be utilized directly
as mRNA (positive stranded RNA viruses) vs those that
require a virion-associated “transcriptase” to produce
mRNAs (negative stranded RNA and dsRNA viruses)

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 Baltimore Classification-
• Positive-sense rna viruses vs other rna viruses
• All RNA viruses must encode their own polymerase
because cells do not have RNA-dependent RNA
polymerase activity

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Baltimor Classification of animal virus
 Major diseases caused by human viruses
"Group" Family Human pathogens (disease)
dsDNA
Poxviridae Variola (smallpox); Orf (pustular dermatitis); Molluscum contagiosum (pustular dermatitis)
Herpesviridae Herpes simplex 1,2 (oral, genital herpes); Varicella-zoster (chickenpox); Epstein-Barr (mononucleosis);
Cytomegalovirus (neonatal abnormalities); HHV6 (roseola); HHV8 (Kaposi's sarcoma)
Adenoviridae Adenovirus (respiratory infection, conjunctivitis)
Polyomaviridae Polyomavirus (benign kidney infection, respiratory disease, leukoencephalopathy)
Papillomaviridae Papillomavirus (warts, genital carcinoma)
ssDNA
Anellovirus Unknown
Parvoviradae B-19 (fifth disease, fetal death)
Retro
Hepadnaviridae Hepatitis B ("serum" hepatitis)
Retroviridae HIV (aids); HTLV (leukemia)
dsRNA
Reoviridae Rotavirus (infantile gastroenteritis)
ssRNA (-)
Rhabdoviridae Rabies virus (rabies)
Filoviridae Ebola virus (ebola)
Paramyxoviridae Parainfluenza virus (respiratory infection); Mumps virus (mumps);
Respiratory syncytial virus (respiratory infection); Measles virus (measles)
Orthomyxoviridae Influenza virus (influenza)
Bunyaviridae Hantaan virus (hemorrhagic fever with renal syndrome)
Arenaviridae Lassa fever virus (hemorrhagic fever)
Deltavirus Hepatitis D (fulminant acute hepatitis)
ssRNA (+)
Picornaviridae Poliovirus (polio), rhinovirus (URI), Hepatitis A ("infectious" hepatitis)
Calciviridae Norwalk (gastroenteritis)
Hepevirus Hepatitis E (acute hepatitis)
Astroviridae Astrovirus (gastroenteritis)
Coronaviridae Coronavirus (respiratory infection)
Flaviviridae Yellow fever virus (yellow fever); Hepatitis C (hepatitis)
Togaviridae Eastern Equine encephalitis virus (encephalitis); Rubella virus (rubella) 51
Virus infectious cycle

Steps in Virus Replications

1. Adsorption or attachment

• Reactive sites on viruses surface interact with specific

receptors on susceptible host cells

• Receptors on the virus capside or envelope irreversibly

binds to cellular receptors on the cell member

► Limit the host spp. & cells infected

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 Introduction…
2. Penetration
• The coat of enveloped viruses fuse with host cell
member
& release the virus nucleo- capsid into host cytoplasm
• Other viruses enter into cell by endocytosis

3.Uncoating

• Viral capsid is broken by viral or cellular enzyme;

• Viral NA is released;

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 Introduction…
• Viral NA transported to within the host cell

► Transcribed to form new progeny virions

4. Biosynthesis or genomic activation

• m-RNA transcribes from viral DNA or; formed directly from

some RNA viruses & codes for viral proteins such as:

 Capsid / Envelope: - Encode structural proteins

- Are building blocks of virion

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 Introduction…
 Enzymes encode for:

- DNA / RNA polymerase

- Other replication enzymes

 NA replication produces new viral genomes

Note:

-Except Poxviruses, DNA viruses replicate in the nucleus;

- RNA viruses mainly in cytoplasm 55

 Introduction…
5. Assembly

• Assembly of viral nucelo - capsid may take place in:

A) Nucleus: E.g: Herpes virus, Adeno virus & others

B) Cytoplasm: E.g: Poliovirus

C) At the cell surface E.g: Influenza virus
N.B: Accumulation of virons at sites of assembly may form

inclusion bodies
► Visible in stained cells with light microscope
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 Introduction…

6. Release

• Release of new intact infectious virions

• May occur by:

 Budding E.g: Enveloped viruses

 Lysis of infected host cells/ tissues

(See diagram )
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 ANTIVIRAL CHEMOTHERAPY

• Obligate intracellular parasites

• replicate intracellularly and often employ host cell
enzymes,macromolecules, and organelles for synthesis of viral
particles. Therefore,
useful antiviral compounds must discriminate between host
and viral functions with a high degree of specificity;

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 ANTIVIRAL CHEMOTHERAPY

• Many viruses encode activities (virulence factors) that

promote the efficiency of viral replication, viral transmission,
the access and binding of the virus to target tissue, or escape
of the virus from host defense and immune resolution.
• Loss of virulence factors results in attenuation of the virus.
• Viruses also encode enzymes that are not present in un-
infected cell.
• These enzymes are critical to viral replication but unnecessary
for cellular function. It is now possible to find specific
inhibitors for viral growth.
 Viruses-contd.
• Many rounds of virus replication take place
during IP ↓
• Virus spreads before symptoms appear ↓
anti viral drug ↓
Relatively ineffective

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Anti-Viral Chemotherapy
Bacteria
Many antibiotics
Highly selective

Viruses
Use host cell metabolism
Selectivity difficult
Toxicity
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 Antiviral drugs
• Needed for viruses for which
No vaccine –availability or not highly
effective(rhinoviruses-serotypes,constantly
changing-influenza / HIV
• To reduce
• Morbidity and mortality
• Immunosuppressed patients

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• DRUG TREATMENT FOR VIRAL INFECTIONS Specific
antiviral drugs have revolutionized
treatment of herpes and HIV infections.
• Effective drugs have also been developed for
influenza A and moderately effective drugs for
respiratory syncytial virus and HCV infections.
• However,the emergence of drug-resistant strains in
treated patients can limit therapeutic efficacy.

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 Anti-Viral Chemotherapy
Reasons for
At present nocontinuing search
drug completely for anti-virals
suppresses viral replication
(with vaccines:
versus possible exception of anti-HIV protease inhibitors)

For many established diseases there is still no effective

vaccine
Rapid mutation (retroviruses)
Or there are problems with the current vaccine
Reassortment (influenza)
New and emerging diseases - no vaccine available
Vaccine development takes many years
Disease that involve immunosuppression (AIDS, cancer,
transplantation) 65
The greatest success in antiviral chemotherapy has been achieved by using:
• inhibitors of nucleic acid synthesis.
• The stages of attachment of virus to host cell
• Stage of uncoating of the viral genome
• Reverse transcription of certain viral genomes
• Regulation of viral transcription
• Replication of viral nucleic acid
• Translation of viral proteins
• Assembly
• Maturation
• Release of progeny virus
→ The mechanism of action varies among anti-virals.
Antiviral agents and their targets

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Antiviral agents ….

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 Interferon
• IFN →binds → cell surface receptors →
block synthesis of several enzymes →block
viral replication inhibiting translation of
mRNA→ protein synthesis
• Problems in clinical use of Interferon
• High doses of interferon required
• Cost
• Given prior to virus exposure or sypmtoms

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• Amantadine and rimantadine
Specific for Influenza by blocking viral
uncoating
Foscarnet : Inhibit viral polymerases and RT
Methiasazone: Historical for inhibitor of
poxviruses, First antiviral agent

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 Examples of Targets for Antiviral Agents
Replication target Agent
• Attachment Peptide analogues of attachment proteins
Neutralizing antibodies
Dextransulfate, Heparin

• Penetration and uncoating Amantadine, rimantadine

• Transcription Interferons

• Protein synthesis Interferon

• DNA replication Nucleoside analogues

(polymerase)

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• Genome replication Nucleoside analogues :Acyclovir, Ganiclovir

• Gene Expression Less amenable

• Assembly / Maturation / Release Relenza taken as an aerosol and

Tamiflu as pill in Influenza
• Virion integrity Nonoxynol-9

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 Viral vaccines
Purpose of viral vaccines is to utilize immune response
of the host to prevent viral disease

Several remarkably effective

– Small pox eradicated
– Reducing annual incidence several viral diseases

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 How Do Vaccines Work?
• Stimulates adaptive immune response
 Natural infection prevents reoccurrence of disease
 Humoral & Cellular responses

• Vaccines prevent or modify disease. Most do NOT

prevent infection.
• Herd immunity reduces spread of disease.
 Disease agents require a certain level of transmission
to be maintained

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 Requirements of a Vaccine
To be effective a vaccine should be capable of eliciting the
following ;-
• Activation of Antigen-Presenting Cells to initiate antigen processing
and producing interleukins.
• Activation of both T and B cells to give a high yield of memory cells.
• Generation of Th and Tc cells to several epitopes, to overcome the
variation in the immune response in the population due to MHC
polymorphism.
• Persistence of antigen, probably on dendritic follicular cells in
lymphoid tissue, where B memory cells are recruited to form
antibody-secreting cells that will continue to produce antibody.
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 Presently available Viral vaccines
• Common Hepatitis A, Hepatitis B, Influenza A
and B, Measles, Mumps, Pliovirus, rabies,
Rubella, Varicella zoster

• Special situations: Adenovirus, Japaneses B

encephalitis, yellow fever

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Viral Diseases Without Vaccines
• HIV
• Smallpox
• Dengue
• RSV / PIV / Metapneumovirus
• Ebola
• West Nile
• SARS
• Rotavirus
• Hepatitis C
• Avian Influenza

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 Types of viral Vaccines
 Live whole virus vaccines
 Killed whole virus vaccines
 Subunit vaccines;- purified or recombinant viral antigen
 Recombinant virus vaccines
  Anti-idiotype antibodies
 DNA vaccines

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 LIVE ATTENUATED VACCINES

Ideal Condition:
Virus alive and fully immunogenic but no virulence

Utilizes virus mutants restricted in some steps in

pathogenesis of disease

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 Live Vaccines
• Attenuated strains

1. Use of a related virus from another animal

2. Administration of pathogenic or partially attenuated

virus by an unnatural route

3. Passage of the virus in an "unnatural host" or host cell

Development of temperature sensitive mutants (used in

conjunction with the above mentioned methods)

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Live Attenuated Virus Vaccines
• Methods of Attenuation:
– repeated passage in a different host
– repeated passage in cold
– Reassortment with attenuated genes

• Mechanisms of attenuation:
– receptor interaction with host cell
– gene expression and replication
– virion mutation

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 Live Attenuated Virus Vaccines
“Weakened” virus which replicates sufficiently in the host to
induce a protective immune response without causing disease

Rotavirus (1998-1999; 2006)

CA-Influenza (2003)
Varicella (1995)
Measles
Mumps MMR (1971)
Rubella
Polio-Sabin (1960)
Adenovirus
Yellow Fever
Smallpox
viral vaccines- dp monga 83
 Live Attenuated Virus Vaccines
• Advantages
– Stimulates a broad immune response
• neutralizing antibody
• secretory IgA for mucosal tissues.
• Cell mediated immunity (CTL)
– All antigens are expressed
– Production costs are lower

• Disadvantages
• Potential for genetic instability
• Risk of reversion to greater virulence in host
– Potential for contamination
– Infection can persist or be more severe in the
immunocompromised.

84
 Potential safety problems --
Live vaccines
1. Underattenuation
2. Mutation leading to reversion to virulence
3. Preparation instability
4. Contaminating viruses in cultured cells
5. Heat liability-storage & storage problems
6. Should not be given to immunocompromized or
pregnant patients

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 Inactivated whole virus vaccines

• Easiest preparations to use

• Simply inactivated
• The outer virion coat should be left intact but the
replicative function should be destroyed.
• must contain much more antigen than live vaccines
• Extreme care –no residual live virulent virus ij vaccine

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 Inactivated whole virus …

• Inactivation by heat or chemicals

Chemicals: formaldehyde or beta- propiolactone

Caution: Excessive treatment can destroy immunogenicity

whereas insufficient treatment can leave infectious virus

capable of causing disease.

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Inactivated Whole Virus Vaccines

Polio-Salk (1955)
Influenza
Hepatitis A
Rabies (1980)
Japanese Encephalitis

88
 Live vs Dead vaccines

Feature                 Live         Dead

 Dose low high
 no. of doses           single         multiple
 need for adjuvant     no               yes
 Duration of immunity   many years   less

 antibody response      IgG          IgA, IgG

 CMI                       good             poor

 Reversion to virulence   possible not possible
89
 Subunit Vaccines
• Now possible to purify virus and viral antigens ↓
identify the peptide sites encompassing the major
antigenic sites of viral antigens ↓
highly purified subunit vaccines can be
produced
High purification → loss of immunogenicity ↓
coupling with immunogenic carrier protein or
adjuvant
Exapmles:Influenza A and B, and HBsAg derived from
the plasma of carriers.  

90
 Synthetic Peptides
• Defined protective viral antigen / s
• Example: Foot and mouth disease in which linear
sequence of 20 amino acids are used
• Identification of immunogenic sites needed
• antigens are precisely defined and free from
unnecessary components which may be associated
with side effects. They are stable and relatively cheap
to manufacture

91
 Recombinant viral proteins
Recombinant hepatitis B vaccine is the only recombinant vaccine
licensed at present.
• Alternate to recombinant vaccines are hybrid virus vaccines
e.g. vaccinia; the DNA sequence coding for the foreign gene is
inserted into the plasmid vector along with a vaccinia virus
promoter and vaccinia thymidine kinase sequences ↓
• The resultant recombination vector is then introduced into
cells infected with vaccinia virus to generate a virus that
expresses the foreign gene ↓
• The recombinant virus vaccine can then multiply in infected
cells
• The genes of several viruses can be inserted, so the potential
exists for producing polyvalent live vaccines. HBsAg, rabies,
HSV and other viruses have been expressed in vaccinia.

92
 Virus host interaction & viral pathogenesis

Effects of viral infection on host cell

• On entry in to the body:

 The virus may replicate and remain at the primary site

 May disseminate to other tissues via the blood stream

or the mononuclear phagocyte and lymphatic system
 Or may disseminate through neurons.

• The blood stream and the lymphatic system are the

predominant means of viral transfer in the body.
 Effects of viral infection…

• The transport of virus in the blood is termed viremia.

• The virus may be either free in the plasma or may be cell

– associated in lymphocytes or macrophages.
• Replication of a virus in macrophages, the endothelial
lining of blood vessels, or the liver can cause the
infection to be amplified and initiate the development of
secondary viremia.
• In many cases, a secondary viremia precedes delivery
of the virus to the target tissue (e.g. liver, brain, skin)

94
Determinants of viral disease
A. Nature of the disease and target tissue
 Portal of entry of virus
 Access of virus to target tissue

 Tissue tropism of viruses

 Permissiveness of cells for viral replication

 Viral pathogen (strain)

B. Severity of disease and Cytopatic ability of virus
• Immune status
– Competence of the immune system & prior immunity to
the virus
• Immune pathology

• Virus inoculum size

• Length of time before resolution of infection
• General health of the person

– Nutrition & other disease influencing immune status

• Genetic make up of the person & age
C. Interaction of virus with target tissue
• Access of virus to target tissue
– Stability of virus in the body
 Temperature
 Acid and bile of the GIT

– Ability to cross skin or mucous epithelial cells (e.g. cross the GIT
in to the blood stream)
– Ability to establish viremia
• Ability to spread through the RES
• Target tissue
 Specificity of viral attachment proteins.
 Tissue –specific expression of receptors
D. Cytopathologic activity of the virus
• Efficiency of viral replication in the cell
 Optimum temperature for replication
 Permissiveness of cell for replication
• Cytotoxic viral proteins

• Inhibition of cell’s macromolecular synthesis

• Accumulation of viral proteins and structures (inclusion
bodies)
• Altered cell metabolism (e.g. cell immortalization)
E. Host protective responses
• Antigen – non specific antiviral responses
 Interferon

 Natural killer cells and macrophages

• Antigen – specific immune responses
 T- Cell response
 Antibody Responses

• Viral mechanisms of escape of immune responses

 Viral pathogenesis
Cytopathogenesis
Three potential outcomes from a viral infection of a cell:
 Abortive infection (failed infection)
 Lytic infection (cell death)
 Persistent infection (infection without cell death)

Persistent infection include:

• Chronic(Non-Lytic or productive) infection

• Latent(Limited viral macromolecular but no virus synthesis)infections

• Recurrent infections
• Transforming(Immortalizing)infections

100
 Productive, Abortive, and Latent Infection

• Infection is the process by which a virus

introduces its genome into a cell. Infection is
productive if new infectious virus is made and
abortive if no new infectious virus is produced
• Infection is latent if the production of
infectious virus does not occur immediately,
but the virus retains the potential to initiate
productive infection at a later time.

101
• The process of re-initiating a productive
infection cycle from the latent state is termed
reactivation.
• Latency is not merely a slow, productive
replication cycle; latency represents a unique
transcriptional and translational state of a
virus in which the productive replication cycle
is not operative but can become operative
when the need arises.
• A cell is permissive if it can support
productive infection, and not permissive if
infection cannot occur at all or is abortive. 102
 Viral Persistence: Chronic & Latent Virus Infections

• Infections are acute when a virus first infects a

susceptible host.
• Viral infections are usually self-limiting.
Sometimes, however, the virus persists for
long periods of time in the host.
• Long-term virus–host interaction may take
several forms.

103
• Chronic infections (also called persistent
infections) are those in which replicating virus
can be continuously detected, often at low
levels; mild or no clinical symptoms may be
evident.
• Latent infections are those in which the virus
persists in an occult (hidden or cryptic) form
most of the time when no new virus is
produced.
104
• There will be intermittent flare-ups of clinical
disease; infectious virus can be recovered
during flare-ups.
• Viral sequences may be detectable by
molecular techniques in tissues harboring
latent infections.
• Inapparent or subclinical infections are those
that give no overt sign of their presence.

105
 Viral pathogenesis…..

Host cell permissiveness to virus

• Non-permissive cell: not allow replication of a particular
type or strain of virus
• Permissive cell: cell provides the biosynthetic machinery
to support complete replicative cycle of virus
• Semi-permissive cell: cell may be very inefficient or
support some but not all steps of virus replications

106
Effect of virus on host cells
A. Direct and indirect damage
– Some viruses are able to shut down host
macromolecules synthesis and thus directly
damage host cells. The damaged cells lyse, which is a
cytopathic effect that can be detected in the laboratory.
– Some virus infections indirectly affect the function of
tissues or organs in the host. e.g. the influenza virus
damages the respiratory epithelium and ciliary’s activity
is severely affected. This results in the accumulation of
bacteria that normally would be eliminated by ciliary’s
action.
B. Inclusion body formation
• The replication of virus in the cytoplasm or nucleus of
infected cells often results in the accumulation of viral as
well as cellular products. These accumulations, which
may be nucleic acids, proteins, and so on, can be stained
and are referred to as inclusion bodies. Some inclusion
bodies are so distinctive that their presence is diagnostic.
• Example

Negribodies- rabies virus.

Owl’s eye-Cytomegalovirus

Cowdry type A-Herpes simplex virus

C. Cell fusion (syncytia formation)
• Enveloped viruses release specific proteins that become
incorporated in to cytoplasmic membrane of the infected cell.
These proteins act as magnets on the infected cell and attract
uninfected cells to their surface. This results in infection of the
originally un infected cell.
• Repetition of this process results in the aggregation of several
infected cells. These aggregated cells eventually fuse,
producing a giant multinucleated cell or syncytium
D. Changes in the surface antigens
• Viruses insert antigens in to the cell membrane of
infected cells.
• These antigens make the cell a target to immunological
destruction by virus – specific antibodies

110
 Host defense against Viral infection

Non-specific Immune defense

• Provide a local rapid response
• Activate the specific immune defense
• Often sufficient to control viral infection
Body temperature and fever
Interferon and other cytokines
Mononuclear phagocytice system
NK-cells

111
Host defense against Viral infection……

• Body temperature and Fever: Limit replication &

destabilized some virus
• Mononuclear phagocytice system:
phgocytized viral & cell debris from virus infected cells
filter many virus from the blood
Macrophage-present antigen to T-cells
• NK-cells_kill virus infected cells

112
 Host defense against Viral
infection……
• Interferon
the body first active defense
Interfere replication
Initiate an anti-viral state in cells
Block viral protein synthesis
Inhibit cell growth
Interferon alpha and beta activate NK cells
Interferon alpha and gamma activate macrophage
Increase MCH antigen expression
Regulate activities of T cells
113
 Host defense against Viral infection……

Specific host defense

• Antibody mediated
Neutralize extra cellular virus
Block viral attachment proteins
Destabilizes viral structure
opsonizes viruses for phagocytosis
promote killing of target cells by the complement &ADDC

Antibody resolves viral infections

Antibody blocks viremic spread to target tissue

114
• Cell mediated
Essential for controlling enveloped &non cytolitic
infection
Recognizes viral peptides presented by MCH molecules
on cell surfaces
CTLs(CD8) kills virus infected cells & as result eliminate
the source of new virus
It respond to viral peptide-class I MCH protein complex
CD4(TH cells) important for the maturation of antibody
response
respond to viral peptide-class II MCH protein complex

115
 Epidemiology of viruses

• Infection of a population is similar to infection of a person, in

that the virus must spread through the population and is
controlled by immunization of the population. To endure
(survive), viruses must continue to infect new,
immunologically naïve, susceptible hosts.
4.1. Exposure
• People are exposed to viruses through out their lives.
• Some situations, vocations, lifestyles, and living arrangement increase the
likelihood that a person will come in contact with certain viruses.
• Many viruses are ubiquitous (found every where), as borne out by the fact that
evidence of exposure (antibodies to the virus) can be detected in most young
children (HSV –1, HHV6, varicella – zoster virus, parvovirus B19) or early adult
hood (EBV and respiratory and enteric viruses).
• Poor hygiene and crowded living, school, and job conditions promote exposure
to respiratory and enteric viruses.
• Day care centers are consistent sources of viral infections, especially viruses
spread by the respiratory and fecal-oral routes.
• Travel, summer camp, and vocations that bring people in
contact with a virus vector such as mosquitoes, put them at
particular risk for infection by arbo-viruses and other
zoonoses.
• Sexual promiscuity also promotes the spread and acquisition
of several viruses.
• Health care workers, such as physicians, dentists, nurses, and
technicians, are frequently exposed to respiratory and other
viruses but are uniquely at risk for acquiring viruses from
contaminated blood (HBV, HIV) or vesicle fluid (HSV).
4.2. Transmission of viruses
• The route of transmission depends on the source of the virus
(the tissue site of viral replication and secretion) and the
ability of the environment and the body route to the target
tissue.
• Non-enveloped viruses (naked viruses) can withstand drying,
the effect of detergents, and extremes of PH and temperature.
• Non-enveloped viruses are generally transmitted by the
respiratory and fecal-oral routes and can often be acquired
from contaminated objects.
• Unlike the non-enveloped viruses, enveloped viruses
are comparatively fragile. They require an intact
envelope for infectivity. These viruses must remain wet
and are spread:

A. In respiratory droplets, blood, mucus, saliva, or semen.

B. By injection

C. By organ transplantation
• Animals can also act as vectors that spread viral disease
to other animals or humans.
• Animals can also be reservoirs.
• Viral diseases that are shared by animals or insects and
humans are called zoonoses.

• Arthropods, including mosquitoes, ticks, and sand f lies

can act as vectors for toga viruses, flavi-viruses, bunya
viruses, and reo-viruses. These viruses are often
referred to as arbo-viruses because they are

arthropod born.
• Other factors that can promote the transmission of
viruses are the potential for asymptomatic infection,
crowded living conditions, certain occupations, certain
lifestyle, day care centers, and travel.
• The persistence of a virus in a community depends on
the availability of a critical number of immunologically
naïve (sero negative), susceptible people.
• Immunization produced by natural means or by
vaccination, is the best way of reducing the number of
such susceptible people.
• The age of the person is an important factor in
determining his or her susceptibility to viral infection.
• Infants, children and the elderly are susceptible to different viruses.
• The competence of a person’s immune response and his or her
immune history determine how quickly and efficiently the infection is
resolved and can also determine the severity of symptoms.
• The geographic distribution of a virus is usually determined by
whether the requisite cofactors or vectors are present or whether
there is an immunologically naïve, susceptible population.
• Seasonal difference in the occurrence of viral disease corresponds
with behaviors that promote the spread of the virus.
• Out breaks of a viral infection often results from the introduction of a
virus (such as hepatitis A) in to a new location.
• The out breaks originate from a common source (e.g. food
preparation)
• Epidemics occur over a larger geographic area and
generally result from the introduction of a new strain of
virus in to an immunologically naïve population.
• Pandemics are worldwide epidemics, usually resulting
from the introduction of a new virus (example, HIV).
• The spread of a virus can be controlled by quarantine,
good hygiene, changes in lifestyle, elimination of the
vector, or immunization of the population.
• Quarantine is restriction of freedom of movement of
apparently well individuals who have been exposed to
infectious disease, which is imposed for the maximal
incubation period of the disease.
• The proper sanitation of contaminated items and
disinfections of the water supply are means of limiting
the spread of enteric viruses.
• The best way to limit viral spread, however, is to
immunize the population.
 Summary: structure,
classification & replication
• Structure
– Nucleic acid in a protein shell, +/- lipid envelope
– Structure impacts on biological properties
• Classification
– Many virus families, organized by structure and biology
• Replication
– Generic scheme
– Varying strategies depending on nucleic acid
 Summary: Pathogenesis

• Cycle of infection
• Effects on cells
– Abortive, lytic, persistent, latent, transforming
infections
• Effects on the organism