Retroviruses

Prepared By Wondmagegn D.
(Assistant Professor)

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 Outline
• Introduction
• Structure and
genomic organization
• Replication Cycle
• Epidemiology
• Clinical Spectrum
• Diagnosis
• Treatment
• Prevention and Control
• Summary

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 Introduction
• The retroviruses are probably the most studied group of
viruses in molecular biology.

• Enveloped, positive-strand ribonucleic acid (RNA) viruses

with a unique morphology and means of replication.

• Retroviridae are distinguished from all other RNA viruses by

the presence of an unusual enzyme, reverse transcriptase.

• These are RNA viruses that belong to family Retroviridae

(Re=Reverse, tr=transcriptase)..

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• The retroviruses are classified by
– the diseases they cause,
– tissue tropism and host range,
– virion morphology, and
– genetic complexity.
• The three subfamilies of human retroviruses are the
– Oncovirinae (HTLV-1, HTLV-2, HTLV-5),
– the Lentivirinae (HIV-1, HIV-2), and
– the Spumavirinae

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• The oncoviruses include the only retroviruses that can
immortalize or transform target cells.
– categorized by the morphology of their core and capsid as
type A, B, C, or D.

• The lentiviruses are slow viruses associated with neurologic

and immunosuppressive diseases.

• The spumaviruses, represented by a foamy virus, cause a

distinct cytopathologic effect but, as already noted, do not
seem to cause clinical disease.

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• Replication proceeds through a DNA intermediate
termed the provirus.

• The provirus integrates randomly into the host

chromosome and becomes a cellular gene.

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 HIV
What is HIV and AIDS??
HIV~ (Human Immunodeficiency Virus)
• is the virus that causes AIDS. Belongs to Lentivirus sub
group of family Retroviridae.
• It harms the body’s immune system by attacking certain
cells, known as helper T cells or CD4 cells. A weakened
immune system is not able to defend the body against
illnesses.

AIDS~ (Acquired Immune Deficiency Syndrome)

• It is related to HIV, but they are not one in the same.
• It is the name given for a variety of
disease manifestations caused by HIV infection.
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 Cont…
• AIDS was first reported in the United States in 1981.The
earliest cases of AIDS were seen in large urban centers,
such as Los Angeles, San Francisco, and New York City.

• Clusters of young, male, homosexual patients exhibited

a puzzling complex of symptoms, including
– severe pneumonia caused by Pneumocystis jiroveci
(ordinarily a harmless eukaryotic organism), Kaposi
sarcoma (ordinarily an extremely rare form of cancer),
sudden weight loss, swollen lymph nodes, and general
suppression of immune function.
• This constellation of signs and symptoms associated
with illness came to be known as acquired immune
deficiency syndrome (AIDS).
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 Cont…
• Early attempts at understanding the disease focused
on the possibility of immune suppression induced
by chronic injectable drug use (IDU) or infection.

• Soon, however, cases were reported in

nonhomosexual, non-IDU patients who had received
blood or blood products by transfusion.

• By 1984, AIDS was recognized as an infectious

disease caused by a virus, and eventually HIV was
isolated from AIDS patients.
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• Both HIV-1 and HIV-2 show considerable sequence variability,
– allowing their classification into a number of subtypes with marked
differences in geographical distribution and association with risk groups.
• HIV-1 variants are classified into three genetic groups:
– major (M),
– outlier (O) and
– non-M, non-O (N)
– P group.
• Group M viruses which dominate the pandemic are further classified
into subtypes A, B, C, D, F, G, H, J and K and several ‘circulating
recombinant forms’ (CRF) that comprise more than one subtype,
e.g. CRF01-A/E.

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• Subtype B is most frequently found in western countries,
whereas other genotypes such as C (Africa, parts of Asia), E
(Thailand) and F (South America) are the main subtypes
responsible for the recent epidemic spread.

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 Structure and Genomic Organization
• Despite their wide range of disease manifestations, all retroviruses
are similar in structure, genome organization, and mode of
replication.
• The viral envelope, formed from the host cell membrane, contains
spiked knobs. These consist of
– a transmembrane protein, TM (fusion protein, also called gp41),
– attachment protein SU (gp120) that binds to a cell receptor
during infection.
• The virion has a cone-shaped, icosahedral core containing the
major capsid protein (CA also called p24).
• Between the capsid and the envelope is an outer matrix protein,
MA (p17), which directs entry of the double-stranded DNA
provirus into the nucleus, and is later essential for the process of
virus assembly.
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 Cont…
• There are two identical copies of the positive sense, single-
stranded RNA genome in the capsid (that is, unlike other
viruses, retroviruses are diploid).

• The RNA is tightly complexed with a basic protein, NC (p7),

in a nucleocapsid structure that differs in morphology
among the different retrovirus genera.

• Also found within the capsid are the enzymes reverse

transcriptase and integrase (which are required for viral DNA
synthesis and integration into the host cell chromosome) and
protease (essential for virus assembly).

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 Viral genes and antigens
• The genome of HIV contains the three structural genes
(gag, pol and env) characteristic of all retroviruses, as
well as other nonstructural and regulatory genes
specific for the virus.

• The products of these genes, both structural and

nonstructural, act as antigens.

• Sera of infected persons contain antibodies to them.

• Detection of these antigens and antibodies is of great

value in the diagnosis and prognosis of HIV infections.
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 Cont…
• The classical structural scheme of a retroviral
genome is: 5’LTR-gag-pol-env-LTR 3’

• LTR (long terminal repeat regions):

 Represent the two end parts of the viral genome,
that are connected to the cellular DNA of the
host cell after integration
 Do not encode for any viral proteins

• Genes for additional regulatory and accessory

proteins of diverse function are located between
the pol and env genes.
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 Cont…

• Gag gene codes for core proteins & structural

virion components
• env genes codes for viral envelop glyco-
proteins
• pol gene codes for the reverse transcriptase &
other enzymes

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 HIV-1 genome organization & gene
products

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 Cont…

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 Cont…
gp120
 Outer envelope protein
 Responsible for attaching the virus to CD4
receptor
 Has cytoxic effect on nerve cells
gp 41
 Induces the hydrophobic interaction of viral & host
cell membrane to fuse
 Immuno-suppressive characteristics when released
as a peptide.

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 Replication Cycle
• The first phase of HIV replication, which includes
viral entry, reverse transcription, and integration of
the virus into the host genome, is accomplished by
proteins provided by the virus.

• The second phase of replication, which includes the

synthesis and processing of viral genomes, mRNAs,
and structural proteins, uses the host cell
machinery for transcription and protein synthesis.

• The end result of HIV replication in most cell types

is cell death.
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 Cont…
1. Binding of gp120 to CD4 molecules and depending
on the virus type also binds to the chemokine
receptors:
 CXCR4….........for T- cell tropic strains
 CCR5……………for macrophage tropic strains
N.B: Both macrophage &T-cell tropic viruses replicate
in CD4
2. Fusion of the virus to cell memb Via gp41
– Binding to a coreceptor activates the viral TM
(fusion) glycoprotein, triggering fusion between
the viral envelope and the cell membrane
– Results in uncoating and entery to the host cell
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 Fig . Mechanism of HIV attachment to its receptors on the
host cell. (Source: NIH reagent program)
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 Cont…
3. Reverse transcription of viral RNA:
• After entering the host cell, the HIV RNA is not
translated. Instead, it is transcribed into DNA by
reverse transcriptase an RNA-directed DNA
polymerase that enters host cells as part of the viral
nucleocapsid.
• This process takes place in the cytoplasm, within the
core structure.
• RT transcribes one of the RNA strands into DNA &
simultaneously RNAase H breaks down the 1st RNA
stand.
• The resulting linear molecule of double-stranded
DNA is the provirus (Proviral DNA).
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 Cont…
RT based transpiration
Occurs without proof reading hence:
 Transcription errors are not corrected
 Error rate is one in 10,000 transcription
 Results in mutation, which can cause drug
resistance or immune elimination.

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 Cont…
4. Integration of the provirus into host cell DNA:

• The dsDNA transported from the cytoplasm to

nucleus with the aid of MA (matrix protein).

• There, viral integrase cleaves the chromosomal DNA

and covalently inserts the provirus; the integrated
provirus thus becomes a stable part of the cell
genome

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 Cont…
5. Transcription and translation
• The provirus is transcribed into a full-length mRNA
by the cell RNA polymerase II.
• The genome-length mRNA has at least 2 functions:
– Some copies will be the genomes of progeny virus, and are
transported to the cytoplasm in preparation for viral
assembly.
– Some copies are translated to produce various proteins.
• Protein synthesis takes place:
 1st proteins….…become regulatory proteins
 2nd proteins…..become structural proteins
6. Assembly and maturation of infectious
progeny:
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 Epidemiology
HIV-is a global pandemic

Eastern Europe
& Central Asia
Western Europe 1.2 – 1.8 million
North America 520 000 – 680 000
East Asia & Pacific
790 000 – 1.2 million
North Africa & Middle 700 000 – 1.3 million
East
470 000 – 730 000 South
Caribbean & South-East Asia
350 000 – 590 000 Sub-Saharan Africa 4.6 – 8.2 million
25.0 – 28.2 million
Latin America
Australia
1.3 – 1.9 million & New Zealand
12 000 – 18 000

Adults and children estimated to be living with

HIV/AIDS (2003): 34 – 46 million total
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 Epidemiology of HIV

• There were an estimated 38.0 million people living with HIV at the
end of 2019.
• In 2019, 68% of adults and 53% of children living with HIV globally
were receiving lifelong antiretroviral therapy (ART).
• Due to gaps in HIV services, 690 000 people died from HIV-related
causes in 2019 and 1.7 million people were newly infected.
• In 2012, an estimated 2.3 million people were newly infected with
HIV. 260,000 were under the age of 15.
• Every day nearly 6,300 people contract HIV nearly 262 every hour.
• In 2012, 1.6 million people died from AIDS. 210,000 of them were
under the age of 15.
• Since the beginning of the epidemic, 76 million people have been
infected with the HIV virus and about 33 million people have died of
HIV/AIDS.
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 Transmission
• Sexual contact
 Homo sexual & hetero - sexual exposure
 STIs fuels rate of transmission
• Blood and blood products
 Blood transfusions
 Intravenous drug abusers
 Health care workers

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• Vertical transmission
– 10 - 30% of babies can acquire infection
– Infection may occur:
–In utero……….vertical transmission
–Perinatally….. infected birth canal &
umbilical cord
–Post natally…….via breast feeding
–PMTCT

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-blood
-semen
-vaginal fluid
-breast milk

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 Clinical spectrum
1. HIV infection
2. AIDS related complex include:
 Immunological, dermatological & neurological sins
 Fever
 Weight loss
 Night sweating
 Chronic diarrhea
3. AIDS
 Final stage of infection
 Complete failure of CMI
 Opportunistic infections & tumors
4. Death
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 WHO HIV/AIDS Classification System

Stage I Stage II Stage III Stage IV

Minor Moderate
Asymptomatic Symptoms Symptoms AIDS

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 Opportunistic infections (OIs)

1. Pneumocystis carnii jervocii

2. Toxoplasmosis
– Encephalitis, causing neurological disorders
3. Viral infection
 CMV retinitis…Blurred vision, reduced visual
acuity
 HSV…Herpetic nurolagia of craniospinal
ganglia
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4. Esophageal candidiasis
– Swallowing difficulties
– Nausea & vomiting
N.B: In > 80% of cases, both oral cavity & pharynx are affected
5. Kaposi's sarcoma (KS)
– About 30 - 50% of HIV infected homosexuals develop KS
– HHV 8: Triggers KS
6. Atypical Mycobacterium infection
– Cause Severe OIs in > 90% HIV pts (CD4 cell count < 50
cells /µl)

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7. Cervical carcinoma
– Pathological cyto smears (pap smears) occur 10
to 15 times greater frequency in sero pos.
women than sero neg.
– The problem is aggravated by HPV types 16 &
18
– Regular gynecological examination of HIV
infected women is highly recommended.

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 Laboratory Diagnosis

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• Tests for HIV infection are performed for
one of four reasons:
– (1) to identify those with the infection so that
antiviral drug therapy can be initiated,
– (2) to identify carriers who may transmit infection
to others (specifically blood or organ donors,
pregnant women, and sex partners),
– (3) to follow the course of disease and confirm the
diagnosis of AIDS, or
– (4) to evaluate the efficacy of treatment

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 Laboratory Diagnosis
• The presence of HIV infection in individuals can be
ascertained only through the use of laboratory
tests.
• Specimens include: Whole blood, plasma , serum
• WHO -UNAIDS have established an algorithm for
the use of various tests for:
 Screening
 Surveillance
 Diagnostic purposes

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Diagnosis of HIV infection can be carried out by
detecting any of the following:
– Nucleic acid-based test (PCR for pro-viral DNA
& RT - PCR for viral RNA)
– p24 antigen testing
– Cell culture
– Antibodies to HIV

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• Antibodies to HIV are detectable: Within 4 to 6 weeks of
infection by commonly employed tests

• Once antibodies appear in the blood, they persist for the

lifetime.

• The level of viraemia immediately after the acute primary HIV

infection or the viral load set point, is a predictive marker of
HIV disease progression

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Fig .3 Laboratory markers during HIV-1 disease progression : WHO, 2004
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 HIV diagnosis in infants
• Diagnosis of HIV infection in babies born to HIV-
infected mothers cannot be established by
conventional antibody tests.

• The presence of anti-HIV Ab in the newborn may

not necessarily indicate primary infection:
 It may be due to passive transmission of anti-HIV
Ab from mother to uninfected child.
 These maternal antibodies may persist even up
to 18 months.

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• Hence, diagnosis in children less than 18 months of
age is possible only by the detection of :
 HIV nucleic acids
 Viral culture
 Detection of the p24 antigen

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 Monitoring of ART
• ART aims at :
 Reducing the viral load
 Augmenting the immune response of patients
• Monitoring of patients on chemotherapy is essential
in all infectious diseases
• It is of greater importance in HIV because of:
 Severity of illness
 Potential of the virus to mutate & become
resistant to drugs
 Cost of treatment
• Hence, lab. are bound to play a critical role in the
successful implementation of ART programme
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HIV viral load measurement
• Useful in monitoring treatment
• Requires for establishment of a baseline plasma
viral load before starting ART
• The viral load in the case of successful ART
becomes undetectable in 4 to 6 months of therapy
• A number of assays for viral load are in use,
however
 Need rigorous standardization
 Continued quality assurance

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• The assessment of viral load, though possible, is :
 Very expensive
 Complex
 Sophisticated procedure
 Hence, not recommended by WHO

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 Monitoring of ART

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 CD4+ T cells count:

• Are primary target cells attacked by HIV

• Depleted during the course of the disease
• The utility of CD4+ T cell measurements
involves:
 Considerations for HIV disease staging &
AIDS definition
 Assessment of prognosis
 Design of clinical trials

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Accurate enumeration of CD4+ T cell counts is very
crucial for:
• Monitoring the rate of progression to AIDS, both
for:
- initiating prophylaxis for OIs
- monitoring the impact /effectiveness of ART
• The CD4 number does not diagnose HIV infection.
This is because CD4 + T-cells may be reduced in
other :
 viral infections not associated with HIV
 immune deficiency states e.g.
Agammaglobulinemia
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 Fig: CD4+ T-cell count and viral load during HIV infection

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 CD4 T- cells & HIV
Decrease in CD4 + T-cells is associated with :
• Increased risk for opportunistic infections
• Disease progression
– CD4 counts of < 200 cells /ul are associated with greater
risk for pneumocystis infections.
– CD4 counts <100 cells /ul are associated with greater risk
for Mycobacterial infections.
• CD4 +T-cell count is used for clinical prognosis,
monitoring therapeutics & for inclusion criteria for
ART.

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 Time Course of CD4+ T- cell Decrease
• HIV infection will progress to AIDS at different rates
in different people.
• The CD4 + T-cell count may be used as a tool to
monitor individuals to determine if they are:
 Fast progressors,
 Slow progressors, or
 Non-progressors

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 Clinical Consequences of HIV Infection (Fast Progressor)

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Clinical Consequences of HIV Infection (Slow Progressors)

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Reference range for CD4-positive T-cells
Reference range for CD4 + T-cells varies with:
 patient age
 geographic location
• Adult: 500 -1500 cells/ul (for white population)
• Ethiopian population: 350 -1350 cells/ul
(Aster et al; 1999)

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 Treatment
1. Non- specific
• Vitamins & minerals are essential (supportive care):
• Chronic HIV infection exerts persistent metabolic
strain on the body
• Link b/n serum conc. of vit E & B12 with disease
progression
2. Treatment of OIs
• Bacterial, fungal, viral, protozoal & others

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3. Specific anti -viral agents
RT inhibitors
- Abacavir - Zidovudine
- Didanosine - Zalcitabine
- Lamivudine
Protease inhibitors
- Indinavir - Ritonavir
- Nelfinavir - Ampernavir
- Saquinavir

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Binding and Fusion Inhibitors
• CCR5 inhibitor (maraviroc)
• T-20 (enfuvirtide/Fuzeon)
Integrase Inhibitor
• Raltegravir (Isentress)

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Major side effects
• Hypersensitivity reactions
• Nausea
• Neuropathy
• Rise in transamianases
• Diarrhea
• Disorder of fat metabolism
• Others

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 Prevention & control
 ABC (abstinence, be faithful & use of condom)
 Blood screening
 ART (PMTCT)
 Supportive care
 Avoiding stigma & discrimination
 Education (behavioral change)

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 Orthomyxoviruses
• All known orthomyxoviruses are influenza viruses.
• Contains negative sense single stranded RNA with
eight segments.
• Three immunologic types are known (A, B and C).
• Only influenza A and B viruses cause significant
human disease.
• Only influenza A can be a zoonosis
• Antigenic changes continually occur within the type
A group and to a lesser degree in B group, C appears
antigenically stable.

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• Influenza B and C have limited genetic diversity and occur
almost exclusively in humans, which are the natural animal
reservoir.
• Occasional transmission of influenza B to mammalian species
such as seals and influenza C to pigs has been described.
• The orthomyxoviruses are enveloped and have a segmented
negative-sense ribonucleic acid (RNA) genome.
• The segmented genome … facilitates development of new
strains through mutation and reassortment of the gene
segments
– among different human and animal (influenza A) strains of virus.

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• This genetic instability is responsible for the
– annual epidemics (mutation: drift) and for
influenza A
– periodic pandemics (reassortment: shift) of
influenza infection worldwide.
• the most famous influenza
pandemic(worldwide) is
– the Spanish influenza that swept the world in
1918-1919, killing 20 to 40 million people.
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• Pandemics caused by novel influenza viruses occurred in
1918, 1947, 1957, 1968, 1977, and 2009.
• Fortunately, prophylaxis with vaccines and antiviral drugs is
available.

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 Orthomyxoviruses
Properties
• Single-stranded RNA, Types A, B, C
• Diameter 80 - 120 nm፣ Pleomorphic, spherical,
filamentous particles
• Segmented genome, 8 segments in A and B
• The segmented genome: facilitates the development
of new strains through the mutation and
reassortment of the gene segments among different
human and animal (influenza A) strains of virus.
• Hemagglutinin and Neuraminidase on surface of
virion

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• Influenza is one of the most prevalent and significant
viral infections.
• Prophylaxis in the form of vaccines and antiviral drugs is
now available for people at risk for serious outcomes.

• Influenza viruses cause respiratory symptoms and the

classic flulike symptoms of fever, malaise, headache, and
myalgias (body aches).

• The term flu, however, has been mistakenly used to refer

to many other respiratory and viral infections

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 Influenza virus structure

• Influenza virions are pleomorphic, appearing spheric or

tubular
• The surface of the particle contains three kinds of spike
proteins:
– hemagglutinin (HA),
– neuraminidase (NA), and
– matrix (M2) protein internally lined by the matrix (M1)
protein..

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 Influenza virus structure

• The ribonucleoprotein complex making up the

core consists of
– at least one of each of the eight single-
stranded RNA segments associated with the
nucleoprotein (NP) and
– the three polymerase proteins (PB2, PB1, PA).
• RNA segments have base pairing between their
3´ and 5´ ends forming a panhandle.
• Their organization and the role of NS2 in the
virion remain unresolved.

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 Orthomyxoviruses
HA - hemagglutinin

NA - neuraminidase

helical nucleocapsid (RNA plus

NP protein)

lipid bilayer membrane

polymerase complex

M1 protein
type A, B, C : NP, M1 protein
sub-types: HA or NA protein
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 Influenza virus- RNA segments

• RNA segments encode one or more protein

• Segment
1-PB2
2-PB1
3-PA
4-HA
5-NP
6-NA
7-M - M1, M2
8-NS-NS1, NS2
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 Hemagglutinin HA

• HA binds virus particle to host cell

• promotes fusion of the envelope to the cell
membrane
• Agglutinates erythrocytes
• elicits the protective neutralizing antibody
response
• Mutation-derived changes in HA are responsible
for the minor (“drift”) and major (“shift”) changes
in antigenicity.

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 Neuraminidase NA

• NA functions at end of viral replication cycle

• NA is sialidase enzyme removes sialic acid from
glycoconjugate
• Facilitates release of virus particle from infected
cell surface
• Help virus negotiate throሀgh mucin layer of RT
→ target epithelial cell.

• A target for two antiviral drugs, zanamivir

(Relenza), and oseltamivir (Tamiflu)

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 Influenza A hemagglutinin and neuraminidase subtypes

Fields Virology, 4th ed, Knipe & Howley, eds, Lippincott Williams & Wilkins, 2001, Table 47-1

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 M1, M2, and NP
• The M1, M2, and NP proteins are type specific
and are therefore used to differentiate influenza
A from B or C viruses.
• The M1 proteins line the inside of the virion and
promote assembly.
• The M2 protein forms a proton channel in
membranes and promotes uncoating and viral
release.
• The M2 of influenza A is a target for the antiviral
drugs amantadine and rimantadine.

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 EPIDEMIOLOGY

• TRANSMISSION
• The virus is spread from person- to- person through
respiratory secretions either as droplets (close contact)
or as airborne infection by droplet nuclei suspended in
the air.
• Incubation period 1-3 days

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 Local Practices

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 Avian Influenza – H5N1

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 Pandemic Influenza

• Result of an Antigenic shift

• Absence of natural immunity
• Year Pandemic No. Deaths
• 1918 Spanish Influenza 40-50 million
• 1957 Asian Influenza 2 million
• 1968 Hong Kong Influenza 1 million

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 Pandemic Influenza Future

• All countries will be effected

• Medical supplies will be Inadequate
• Large number of deaths will occur

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 THE IMPACT OF INFLUENZA

• recently some increase in morbidity and

mortality - possible factors?
– more elderly people
– more high risk neonates
– more immunosuppressed patients

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• Strains of influenza A virus are classified by the
following four characteristics:
– Type (A)
– Place of original isolation
– Date of original isolation
– Antigen (HA and NA)
• A/Bangkok/1/79 (H3N2),

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• Strains of influenza B are designated by
– (1) type,
– (2) geography, and
– (3) date of isolation
• (e.g., B/Singapore/3/64), but without specific
mention of HA or NA antigens, because
influenza B does not undergo antigenic shift or
pandemics as does influenza A.

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 Nomenclature
• Name of influenza virus strain includes:
– Type
– Host of origin
– Geographic origin
– Strain number
– Year of isolation
– Antigenic description of HA and NA for type A
• Host of origin not in human isolates

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 Nomenclature

• A/Hong Kong/03/68(H3N2)
• A/swine/Iowa/15/30(H1N1)
• So far 15 subtypes of HA (H1-15)
• Nine subtypes of NA (N1-9)
– Many different combination
– Possible 15 X 9

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 Reservoirs of Influenza Viruses

• Aquatic birds
• Pigs
• Humans

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 Influenza A reservoir
• The reservoir of influenza A
viruses.

• The working hypothesis is that

wild aquatic birds are the
primordial reservoir of all
influenza viruses for avian and
mammalian species.

• Transmission of influenza has

been demonstrated between
pigs and humans (solid lines).

• There is extensive evidence for

transmission between wild
ducks and other species.

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 Antigenic Variation
• Influenza viruses tend to undergo changes from
time to time.
• There are two types of changes:
• (1) antigenic shift,
• (2) antigenic drift.

• These changes in the antigenic characteristics of

influenza viruses determine the extent and
severity of influenza epidemics

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 Antigenic drift
• Minor antigenic changes in HA and NA →
antigenic drift
• This results from mutation in the RNA segments
coding for either the HA or NA→ AA changes in
protein---RNA virus escape recognition of host’s
IR
• immune response no longer protects fully
• sporadic outbreaks, limited epidemics
• This involves no change in serotype; there is
merely an alteration in amino acid sequence of
HA or NA leading to change in antigenicity.

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 Antigenic shift

• This term denotes MAJOR changes in

hemaglutinin and neuraminidase resulting from
reassortment of gene segments involving two
different influenza viruses.
• Genetic reassortment especially doubly infected
cells
• Not in Type B and C
• “new” HA or NA proteins
• When this occurs, worldwide epidemics may be
the consequence since the entire population is
susceptible to the virus.

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 A Pandemic Strain Emergence:
Reassortment of Influenza Viruses
Antigenic Shift

Avian Human
virus virus
Avian
Reservoir

Other
mammals?
New
Swine
Reassorted
virus
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 Influenza replication

From Medical Microbiology, 5th ed., Murray, Rosenthal & Pfaller, Mosby Inc., 2005, Figure 60-2.
8/23/2020 118
 Clinical Feature

• Influenza is an acute respiratory illness

characterized by fever, headache, myalgia,
coryza, sore throat and cough.
• It can cause mild to severe illness, and at times
can lead to death.
• The flu is different from a cold.
• The flu usually comes on suddenly.
• Cough is frequently severe and protracted.
• Duration of illness is usually 2-7 days

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 ‘FLU’

• True influenza
– Influenza virus A or influenza virus B (or
influenza virus C infections - much milder)
• Febrile respiratory disease with systemic
symptoms caused by a variety of other
organisms often inaccurately called ‘flu’.

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• People who have the flu often feel some or all of these
symptoms:
– Fever or feeling feverish/chills
– Cough
– Sore throat
– Runny or stuffy nose
– Muscle or body aches
– Headaches
– Fatigue (tiredness)
– Some people may have vomiting and diarrhea,
though this is more common in children than adults.

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 – very young
– elderly
– immuno-compromised
– heart or lung disease

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 Pulmonary complications

• croup (young children)

• primary influenza virus pneumonia
• secondary bacterial infection
– streptococcus pneumoniae
– staphlyococcus aureus
– hemophilus influenzae

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 Non-pulmonary complications
• Myositis
• Cardiac complications
• Recent studies report encephalopathy
• Liver and CNS
– Reye’s syndrome
• peripheral nervous system
– Guillian-Barré syndrome
_______________________________________________
* Myositis=muscle disease in which
inflammation and degenerative changes occur

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 Pneumonia
• Serious complication
• Elderly, debilitated
• Pregnancy
• Chronic diseases

8/23/2020 125
 MORTALITY

• Major causes of influenza virus- associated death

– Bacterial pneumonia
– Cardiac failure
• 90% of deaths in those over 65 years of age

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 Diagnosis
• Since the clinical picture of influenza is
nonspecific, its specific diagnosis must be
confirmed by laboratory tests.
• Influenza viruses are obtained from respiratory
secretions taken early in the illness.
• Serology antibodies to HA,NA,NP and M during
infection
• ELISA
• PCR
• Rapid tests
• Provisional - clinical picture + outbreak

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 Treatment/prevention
• Immunoprophylaxis with vaccine
• Chemoprophylaxis and chemotherapy
• Amantidine and Rimantidine target matrix;
• Zanamivir and Oseltamivir target NA
• oseltamivir also approved for prophylaxis
• killed and live vaccines need constant
updating

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 Paramyxoviridae

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 Outline

• Introduction
• Structure and replication
• PIV
• Measles Virus
• Mumps Virus
• RSV

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 Introduction

• The family Paramyxoviridae consists of three genera:

1. Paramyxovirus, which includes the parainfluenza viruses
and mumps virus;
2. Pneumovirus, which includes respiratory syncytial virus;
and
3. Morbillivirus, which includes the measles virus.

• A new group of highly pathogenic paramyxoviruses, including

two zoonosis-causing viruses, Nipah virus and Hendra
virus, was identified in 1998 after an outbreak of severe
encephalitis in Malaysia and Singapore. .

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• Their virions have similar morphologies and protein
components.
• Importantly, paramyxoviruses induce cell-to-cell fusion
(syncytia formation and multinucleated giant cells).

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 Cont…
• These agents cause some well-known major
diseases.

• Measles virus causes a potentially serious

generalized infection characterized by a
maculopapular rash ( rubeola).

• Parainfluenza viruses cause upper and lower

respiratory tract infections, primarily in children,
including pharyngitis, croup, bronchitis, bronchiolitis,
and pneumonia.
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 Cont…
• Mumps virus causes a systemic infection whose
most prominent clinical manifestation is parotitis.

• RSV causes mild upper respiratory tract infections in

children and adults but can cause life-threatening
pneumonia in infants.

• Measles and mumps viruses have only one serotype,

and protection is provided by an effective live
vaccine.

• Paramyxoviridae are transmitted in respiratory

droplets and initiate infection in the respiratory tract.
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 Cont…

• Paramyxoviruses earlier grouped with

Orthomyxoviruses because of HA and NA
activities but they do have the following
differences
– RNA non segmented
– RNA is RNase resistant
– Transcription of viral RNA in cytoplasm
– Fusion of virus with cell membrane
– Genetic reassortment-rare

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 Structure and Replication
• Paramyxoviruses are relatively large viruses with a
negative sense, single-stranded ribonucleic acid (RNA)
genome in a helical nucleocapsid surrounded by a
pleomorphic envelope.
• The envelope contains two glycoproteins
• The first, the HN protein (H stands for hemagglutinin,
and N for neuraminidase), is involved in the binding of
the virus to a cell; measles virus lacks the neuraminidase
activity.
• The second, the F protein (F stands for fusion), functions
to fuse viral and cellular membranes, thus facilitating
virus entry into the cytoplasm where viral replication
occurs
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 Cont…

• Envelope Glycoproteins
• HN glycoprotein → HN - hemagglutinin +
neuraminidase activities → Parainfluenza viruses
• Measles - referred to as H protein - no
neuraminidase activity;
• RSV - G protein - neither activity.
• FUSION (F) Protein – infection and pathogenesis-
fusion of viral envelope with plasma membrane
of host cell →cell to cell fusion

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8/23/2020 139
 Cont…

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 Cont…
pleomorphic HN/H/G glycoprotein
SPIKES
F glycoprotein
SPIKES

helical nucleocapsid (RNA plus

NP protein)

lipid bilayer membrane

polymerase
complex

M protein
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 Recent Classification

GENUS GLYCOPROTEINS TYPICAL MEMBERS

PARAMYXOVIRUS SUBFAMILY
Paramyxovirus HN, F HPIV1, HPIV3
Rubulavirus HN, F HPIV2, HPIV4, mumps virus
Morbillivirus H, F measles virus
PNEUMOVIRUS SUBFAMILY
Pneumovirus G, F respiratory syncytial virus
Metapneumovirus G, F metapneumoviruses

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 Virus H N Fusion protein

Measles virus + - +

mumps + + +

RSV - - +

Parainfluenza + + +

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 Replication
• Replication of the paramyxovirus genome, takes place in the
cytoplasm of the host cell.
• Virus attaches to host cell surface receptors through HN, H or
G glycoproteins.
• Fusion with the plasma membrane; ribonucleocapsid is
released in the cytoplasm.
• The RNA of the virus is translated into positive sense RNA
through use of the enzyme RdRp, which is short for RNA-
dependent-RNA-polymerase.
• The positive sense RNA can then act as mRNA which is read
by the host cell, and the host cell’s mechanisms build the
necessary proteins, as well as gets copied into more negative
sense RNA to use in the daughter virus.

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 Cont…

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 Parainfluenza virus
• Four serologic types within the parainfluenza genus are
human pathogens.
• are respiratory viruses that usually cause mild cold like
symptoms but can also cause serious respiratory tract
disease.
• Serologic types 1, 2,and 3 are second only to RSV as
important causes of severe lower respiratory tract
infection in infants and young children.
• They are especially associated with
laryngotracheobronchitis (croup).
• Type 4 causes only mild upper respiratory tract infection
in children and adults.
• limited to U.R.T. (no viremia)

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 Cont…

• Transmission
– Aerosols-Person to person contact
• IP 2-6 days
• Wide distribution
• Type 3 most prevalent- pneumonia infants
• Type 1 and 2 – croup infants
• Most children get infection-no serologic
variation-rare infection in adults.

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 Pathogenesis
• Infect epithelial cells of URT
• The virus replicates more rapidly than measles and
mumps viruses and can cause giant cell formation
and cell lysis.
• Unlike measles and mumps rarely viremia
• The viruses generally stay in the upper respiratory
tract, causing only cold like symptoms.
• In approximately 25% of cases, the virus spreads to
the lower respiratory tract, and in 2% to 3%, disease
may take the severe form of
laryngotracheobronchitis.

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 PIV Epidemiology

• Parainfluenza viruses are ubiquitous, and

infection is common.
• The virus is transmitted by person to person
contact and respiratory droplets.
• Primary infections usually occur in infants and
children younger than 5 years.
• Reinfections occur throughout life, indicating
short-lived immunity.
• Type 3 most prevalent

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 Clinical Manifestations

• Croup (laryngotraheobroncitis) - most common

manifestation of parainfluenza virus infection.
However other viruses may induce croup e.g.
influenza and RSV.
• Other conditions that may be caused by
parainfluenza viruses include Bronchiolitis,
Pneumonia, Flu-like tracheobronchitis, and
Corza-like illnesses.

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 PIV Recovery

• All infants- antibodies but not protective

• Immunity short lived
• Cell mediated immunity both cell damage and
confers protection
• IgA protective but short lived
• Antibodies to F Neutralize prevent cell to cell
spread
• Multiple serotypes and short live immunity ↓
– Reinfection more common →milder
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 Laboratory Diagnosis

• Detection of Antigen - a rapid diagnosis can be made by

the detection of parainfluenza antigen from
nasopharyngeal aspirates and throat washings.

• Virus Isolation - Parainfluenza virus is isolated from nasal

washings and respiratory secretions and grows well in
primary monkey kidney cells.

• The presence of virus-infected cells in aspirates or in cell

culture is indicated by the finding of syncytia and is
identified with immunofluorescence.

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 Laboratory Diagnosis

• Serology - a retrospective diagnosis may be

made by serology. CFT most widely used.

• Rapid RT-PCR techniques are becoming the

method of choice to detect and identify
parainfluenza viruses from respiratory secretions.

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8/23/2020 156
8/23/2020 157
Infected cell fusion caused by paramyxoviridae

Uninfected Infected
8/23/2020 158
 PIV Treatment

• Ribavirin-promising-aerosol form
• Experimental- killed virus vaccine-good antibody
titer - no protection.
• Subunit vaccines and Live attenuated vaccine-
intranasal being tested.
• Careful monitoring of the upper airway.

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 Measles
• Single serotype & Humans are the natural host
• Measles virus causes a potentially serious
generalized infection characterized by a
maculopapular rash (rubeola).

• Measles, also known as rubeola, is one of the five

classic childhood exanthems, along with rubella,
roseola, fifth disease, and chickenpox.

• Historically, measles was one of the most common

and unpleasant viral infections, with serious
potential sequelae.

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• Before 1960, more than 90% of the population younger than
20 years had experienced the rash, high fever, cough,
conjunctivitis, and coryza of measles.

• Measles is still one of the most prominent causes of disease

(>10 million cases per year) and death (120,000 deaths in
2012) worldwide in unvaccinated populations.

• The development of effective vaccine programs has made

measles a rare disease in developed countries, but children
remain unvaccinated or do not receive their boosters and
outbreaks of measles occur.

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 Measles

8/23/2020 162
 Disease Mechanisms of Measles Virus

• Virus infects epithelial cells of respiratory tract.

• Virus spreads systemically in lymphocytes and by viremia.
• Virus replicates in cells of conjunctivae, respiratory tract,
urinary tract, lymphatic system, blood vessels, and central
nervous system (CNS).
• Rash is caused by T-cell response to virus-infected
epithelial cells lining capillaries.
• Virus causes immunosuppression.
• Cell-mediated immunity is essential to control infection.
• Sequelae in the CNS may result from immunopathogenesis
(postinfectious measles encephalitis) or development of
defective mutants (subacute sclerosing panencephalitis).

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 Measles pathogenesis

FIGURE 59-4

FIGURE 59-3

8/23/2020 164
 Clinical Syndromes
• Measles is a serious febrile illness .

• The incubation period lasts 7 to 13 days, and the

prodrome starts with high fever and “CCC and P”—
cough, coryza, conjunctivitis, and photophobia.

• The disease is most infectious during this time.

• After 2 days of prodromal illness, the typical mucous

membrane lesions known as Koplik spots appear.

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8/23/2020 166
 Measles rash

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8/23/2020 168
 Laboratory Diagnosis

• The clinical manifestations of measles are usually so

characteristic that it is rarely necessary to perform
laboratory tests to establish the diagnosis.
• The measles virus is difficult to isolate and grow,
although it can be grown in primary human- or monkey-
cell cultures.
• Respiratory tract secretions, urine, blood, and brain
tissue are the recommended specimens.
• It is best to collect respiratory and blood specimens
during the prodromal stage and until 1 to 2 days after
the appearance of the rash.

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 Laboratory Diagnosis

• Measles antigen can be detected in pharyngeal cells or

urinary sediment with immunofluorescence;
• The measles genome can be identified by RT-PCR.

• Characteristic cytopathologic effects, including

multinucleated giant cells with cytoplasmic inclusion
bodies, can be seen in Giemsa stained cells taken from
the upper respiratory tract and urinary sediment.

• Antibody, especially immunoglobulin M (IgM), can be

detected when the rash is present.

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Treatment, Prevention, and Control
• a live attenuated measles vaccine
• Because measles is strictly a human virus with
only one serotype,
– it is a good candidate for eradication,
– but this is prevented by difficulties in distributing
the vaccine to regions that lack proper refrigeration
facilities (e.g., Africa) and distribution networks.

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 MUMPS

• Mumps virus is the cause of acute, benign viral

parotitis (painful swelling of the salivary glands).
• H and N + fusion protein on envelope spikes
• Humans are the natural host.
• Thermo labile
• Mumps is rarely seen in countries that promote
use of the live vaccine, which is administered
with the measles and rubella live vaccines.

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8/23/2020 173
 Mumps – clinical presentation

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8/23/2020 175
8/23/2020 176
 Time course of Mumps infection

FIGURE 59-8

8/23/2020 177
 LABORATORY DIAGNOSIS
• Virus can be recovered from saliva, urine, the pharynx,
secretions from Stensen’s duct, and cerebrospinal fluid.
• Virus is present in saliva for approximately 5 days after the
onset of symptoms and in urine for as long as 2 weeks.
• Mumps virus grows well in monkey kidney cells, causing the
formation of multinucleated giant cells.
• Clinical diagnosis can be confirmed by serologic testing. A
fourfold increase in the virus-specific antibody level or the
detection of mumps-specific IgM antibody indicates active
infection.
• Enzyme-linked immunosorbent assay, immunofluorescence
tests, and hemagglutination inhibition can be used to detect
the mumps virus antigen or antibody.

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 TREATMENT & PREVENTION
• Vaccines provide the only effective means for
preventing the spread of mumps infection.
• Antiviral agents are not available.

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 Respiratory Syncytial Virus
• Most important cause of pneumonia and
bronchiolitis in infants
• Fusion proteins- syncytia formation
• Humans and chimpanzees- natural host
• 2 serotype: A & B
• is a member of the Pneumovirus genus.

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• lacks hemagglutinin and neuraminidase
activities.
• It is the most common cause of fatal acute
respiratory tract infection in infants and
young children.
• It infects virtually everyone by 2 years of
age, and reinfections occur throughout
life, even among elderly persons.

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8/23/2020 182
8/23/2020 183
8/23/2020 184
 LABORATORY DIAGNOSIS

• RSV is difficult to isolate in cell culture.

• The presence of the viral genome in infected
cells and nasal washings can be detected by RT-
PCR techniques, and
• commercially available immunofluorescence and
enzyme immunoassay tests are available for
detection of the viral antigen.

8/23/2020 185
 TREATMENT & PREVENTION

• In otherwise healthy infants, treatment is supportive,

consisting of the administration of oxygen,
intravenous fluids, and nebulized cold steam.
• Ribavirin, is administered by inhalation.
• Passive immunization with anti-RSV immunoglobulin
is available for premature infants.
• Infected children must be isolated.
• No vaccine is currently available for RSV prophylaxis.

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Picornaviri
dae
Prepared By Wondmagegn D.(MSC.)

187
Objectives

• Explain the properties of Picornavirus

• Describe the pathogenesis and clinical features
Picornavirus infection
• Illustrate epidemiology Picornavirus
• Describe the diagnosis Picornavirus infection

188
• Picornaviridae is one of the largest families of viruses and
includes some of the most important human and animal
viruses.

• As the name indicates, these viruses are small (pico)

ribonucleic acid (RNA) viruses that have a naked capsid
structure.

• The family has more than 230 members divided into nine
genera including Enterovirus, Rhinovirus, Hepatovirus,
Cardiovirus, and Aphthovirus.

189
• At least 90 serotypes of human enteroviruses exist; they are
members of the polioviruses, group A or B Coxsackie viruses,
or echoviruses.

• Several different disease syndromes may be caused by a

specific serotype of enterovirus. Likewise, several different
serotypes may cause the same disease, depending on the target
tissue affected.

• The enteroviruses are distinguished from the rhinoviruses by

the stability of the capsid at pH 3, the optimum temperature for
growth, the mode of transmission, and their diseases
190
 Cont…
• The capsids of enteroviruses are very resistant to harsh
environmental conditions and the conditions in the GIT,
– facilitates their transmission by the fecal-oral route.

• Although they may initiate infection in the gastrointestinal

tract, the enteroviruses rarely cause enteric disease.

• Coxsackieviruses are named after the town of Coxsackie,

New York, where they were first isolated.
– They are divided into two groups, A and B, on the basis of certain
biological and antigenic differences and
– are further subdivided into numeric serotypes on the basis of additional
antigenic differences.

191
• The name echovirus is derived from enteric cytopathic human
orphan
– because the disease associated with these agents was not initially
known.
• Since 1967, newly isolated enteroviruses have been
distinguished numerically.

192
 Cont…
• The human rhinoviruses consist of at least 100 serotypes and
are the major cause of the common cold.

• They are sensitive to acidic pH and replicate poorly at

temperatures above 33° C.

• These properties usually limit rhinoviruses to causing upper

respiratory tract infections.

193
 Structure and properties

• Virion is a naked, small (25 to 30 nm), icosahedral capsid

enclosing a single-stranded positive RNA genome.
• Enteroviruses are resistant to pH 3 to pH 9, detergents, mild
sewage treatment, and heat.
• Rhinoviruses are labile at acidic pH; optimum growth
temperature is 33° C.
• Genome is a messenger ribonucleic acid (mRNA).
• Naked genome is sufficient for infection.
• Virus replicates in cytoplasm.
• Viral RNA is translated into polyprotein, which is then cleaved
into enzymatic and structural proteins.
• Most viruses are cytolytic. 194
 Picornaviridae

• Two genera of Picornaviridae :

• Enteroviruses
• Rhinovirus groups

• Have an identical morphology but can be

distinguished based on clinical, biophysical, and
epidemiological studies.

195
 Picornaviridae

• Enteroviruses
– grow at a wide pH range (i.e, 3-10).
– After initial replication in the oropharynx,
enteroviruses survive the acidic environment
of the stomach.
– The small intestine is the major invasion site of
enteroviruses, which replicate best at 37°C.

196
 Picornaviridae

• Rhinoviruses
– replicate at a pH of 6-8.
– After initial replication in the nasal passages,
the acidic environment of the stomach
destroys rhinoviruses.
– Rhinoviruses optimally replicate at 33°C and
primarily infect the nasal mucosa.

197
 Disease Mechanisms of Picornaviruses

• Enteroviruses enter via the oropharynx, intestinal

mucosa, or upper respiratory tract and infect the
underlying lymphatic tissue; rhinoviruses are restricted to
the upper respiratory tract.

• In the absence of serum antibody, enterovirus spreads

by viremia to cells of a receptor-bearing target tissue.
• Different picornaviruses bind to different receptors, many
of which are members of the immunoglobulin
superfamily (i.e., intercellular adhesion molecule-1).

• The infected target tissue determines the subsequent

198
disease.
 Disease Mechanisms of Picornaviruses

• Viral, rather than immune, pathologic effects are usually

responsible for causing disease.

• The secretory antibody response is transitory but can

prevent the initiation of infection.

• Serum antibody blocks viremic spread to target tissue,

preventing disease.

• Enterovirus is shed in feces for long periods. Infection is

often asymptomatic or causes mild, flulike, or upper
respiratory tract disease.
199
200
201
General characteristics of enteroviruses

1- They are transmitted by the fecal oral route.

2- They are acid stable.
3- They replicate in the pharynx and small
intestine.
4- They cause neurological and non-neurological
diseases.
5- They shed in stool.
6- Do not cause diarrhea.

202
Endemicity

Enteroviruses are endemic in areas with:

• Low standard of hygiene and sanitation.
• Primitive sewage system.
• Absence of proper drinking water-pipe system.
• Low educational level.
• Crowded living condition.

203
 Poliovirus Infections
• There are three poliovirus types, with 85% of the cases
of paralytic polio caused by type 1.

• Reversion of the attenuated vaccine virus types 2 and 3

to virulence can cause vaccine-associated disease.

• Wild-type polio infections are rare because of the

success of polio vaccines

204
 Poliomyelitis

• Poliomyelitis (polio) is a highly infectious viral disease that

largely affects children under 5 years of age.
• The virus is transmitted by person-to-person spread mainly
through
• the faecal-oral route or, less frequently, by a common vehicle
(e.g. contaminated water or food) and
• multiplies in the intestine, from where it can invade the
nervous system and cause paralysis.

205
• Wild poliovirus cases have decreased by over 99% since 1988,
from an estimated 350 000 cases in more than 125 endemic
countries then to 175reported cases in 2019.
• Of the 3 strains of wild poliovirus (type 1, type 2 and type 3),
• wild poliovirus type 2 was eradicated in 1999 and
• no case of wild poliovirus type 3 has been found since the last
reported case in Nigeria in November 2012.
• Both strains have officially been certified as globally
eradicated.
• As at 2020, wild poliovirus type 1 affects two countries:
Pakistan and Afghanistan.

206
207
 Pathogenesis

• Entry through mouth

• Replication in pharynx, GI tract, local lymphatic
• Hematologic spread to lymphatic and central
nervous system
• Viral spread along nerve fibers
• Destruction of motor neurons

208
 Poliovirus Epidemiology

• During epidemic outbreaks, type I is most frequently

isolated (in 65-95 per cent of cases) while types II and III
account for the remaining 5-35 per cent of cases.
• Reservoir Human
• Transmission Fecal-oral
– Ingestion via contaminated food and water
– Contact with infected hands
– Inhalation of infectious aerosols
• Communicability
– 7-10 days before onset
– Virus present in stool 3-6 weeks

209
 POLIO-CLINICAL

• Poliomyelitis is an acute infectious disease that in its

serious form affects the central nervous system. The
destruction of motor neurons in the spinal cord results in
flaccid paralysis (less than 0.1%).
• However, most poliovirus infections are subclinical.
• Incubation Period
3 days to 35 days
Usually 7-14 days
• Asymptomatic infection, to mild fever, to severe and
permanent paralysis

210
• Poliovirus may cause one of the following four outcomes in
unvaccinated people, depending on the progression of the
infection
• Asymptomatic illness results if the viral infection is limited to
the oropharynx and the gut. At least 90% of poliovirus
infections are asymptomatic.

• Abortive poliomyelitis, the minor illness, is a nonspecific

febrile illness occurring in approximately 5% of infected
people. Fever, headache, malaise, sore throat, and vomiting
occur in such persons within 3 to 4 days of exposure.

211
• Nonparalytic poliomyelitis or aseptic meningitis occurs in
1% to 2% of patients with poliovirus infections.
– the virus progresses into the central nervous system and the meninges,
causing back pain and muscle spasms in addition to the symptoms of
the minor illness.

• Paralytic polio, the major illness, occurs in 0.1% to 2.0% of

persons with poliovirus infections and is the most severe
outcome.

212
213
214
Child with polio sequelae
215
216
 Polio- Diagnosis

• Recovery of virus throat swab/rectal swab

• Specimen frozen during transit
• Human / monkey cells
• CPE 3-6 days
• Serology: Very rarely used for diagnosis since
cell culture is efficient Paired serum

217
 Polio- Immunity

• Infection-Permanent type specific

• Low degree heterotypic
• Infants protected by maternal antibodies
• Passive immunity mother to offspring- maternal
ab → disappears 6 months

218
 Treatment, Prevention, and Control

• Pleconaril is available on a limited basis. The drug inhibits

penetration of picornaviruses into the cell. It must be
administered early in the course of the infection.
• The two types of poliovirus vaccine are
– (1) inactivated polio vaccine (IPV), developed by Jonas Salk, and
– (2) live attenuated oral polio vaccine (OPV), developed by Albert
Sabin.
• Both vaccines incorporate the three strains of polio, are stable,
are relatively inexpensive, and induce a protective antibody
response

219
OPV
Advantages
• Effectiveness
• Induces long lasting immunity.
• Induction of secretory antibody response similar to
that of natural infection( gut immunity).
• Possibility of attenuated virus circulating in
community by spread to contacts (indirect
immunization)(herd immunity)
• Ease of administration
• Lack of need for repeated boosters

220
OPV
Disadvantages
• The only disadvantage of this vaccine is the vaccine strain
particular type 3 strain can reverts to virulerence and cause
paralysis in those who just been vaccinated.

• It is estimated that vaccine induced poliomyelitis is seen in

rate of 1 in 3000,000 vaccinations.

• Unsafe administration for immunodeficient patients

221
IPV

Advantages
• Good stability during transport and in storage
• Safe administration in immunodeficient patients
• No risk of vaccine-related disease

222
IPV

Disadvantages
• Lack of induction of local (gut) immunity
• Need for booster vaccine for lifelong immunity
• Fact that injection is more painful than oral administration
• Fact that higher community immunization levels are needed
than with live vaccine

223
 Polio Eradication
• Polio cases have decreased by over 99% since 1988, from an
estimated 350 000 cases then, to 406 reported cases in 2013.
• The reduction is the result of the global effort to eradicate the
disease.
• In 2014, only 3 countries (Afghanistan, Nigeria and Pakistan)
remain polio-endemic, down from more than 125 in 1988.
• In 1994, the WHO Region of the Americas was certified polio-
free, followed by the WHO Western Pacific Region in 2000
and the WHO European Region in June 2002.

224
• On 27 March 2014, the WHO South-East Asia Region was
certified polio-free,

• This achievement marks a significant leap forward in global

eradication, with 80% of the world’s population now living in
certified polio-free regions.

• Of the 3 types of wild poliovirus (type 1, type 2 and type 3),

type 2 wild poliovirus transmission has been successfully
stopped (since 1999).

225
Coxsackieviruses
• The coxsackieviruses comprise a large subgroup of the
enteroviruses.

• They produce a variety of illnesses in human beings,

including aseptic meningitis, herpangina, hand, foot, and
mouth disease, myo- and pericarditis, common colds,
and possibly diabetes.

• Coxsackieviruses are named after the town of Coxsackie,

New York, where they were first isolated.

226
Coxsackieviruses
• They are divided into two groups, A and B, on the basis
of lesions observed in suckling mice, certain biologic and
antigenic differences

• and are further subdivided into numeric serotypes on the

basis of additional antigenic differences.

• coxsackieviruses are distinguished from other

enteroviruses by their pathogenicity for suckling rather
than adult mice.

227
Coxsackieviruses
• Group A viruses produce a diffuse myositis with
acute inflammation and necrosis of fibers of
voluntary muscles.

• Group B viruses produce

– focal areas of degeneration in the brain,
– necrosis in the skeletal muscles, and
– inflammatory changes in the dorsal fat pads,
the pancreas and occasionally myocardium.
228
 Cont…

At least 29 immunological types

• 23 group A and
• 6 group B
• Each has a type specific antigen.

229
 Herpangia

• Severe febrile pharyngitis

• Coxsackie A virus (2-6, 8, 10)
• No relation with herpes virus
• Abrupt onset
• Discrete vesicles on posterior half of palate
• Self limiting-small children

230
 Lab Diagnosis

• Isolation of virus : throat washings and stools ↓

–Tissue culture-CPE 5-14 days/suckling
mice – illness 3-8 days group A 5-14
days group B
• Serology : persist for years
• Serological tests difficult to evaluate –
multiplicity of types

231
 Echoviruses
• Enteric, cytopathic, human, orphan viruses.
• More than 32 serotypes (types 1-34; echovirus
10 and 28) -not all pathogenic
• Causes:-
– Aseptic meningitis
– Rashes types 4,9,16,18
– Conjunctivitis
– Infantile diarrhea
– Muscle weakness etc

232
 Rhinoviruses

• Rhinoviruses are the most important cause of the common

cold and upper respiratory tract infections.
• Such infections are self-limited, however, and do not cause
serious disease.
• More than 100 serotypes of rhinovirus have been identified.
• At least 80% of the rhinoviruses have a common receptor that
is also used by some of the coxsackieviruses.
– ICAM-1, a member of the immunoglobulin superfamily,
– which is expressed on epithelial, fibroblast, and B-lymphoblastoid
cells.

233
 Epidemiology
• Rhinoviruses cause at least half of all upper respiratory tract
infections.
• Other agents likely to cause the symptoms of the common cold
are enteroviruses, coronaviruses, adenoviruses, and
parainfluenza viruses.
• Rhinoviruses can be transmitted by two mechanisms: as
aerosols and on fomites (hands or inanimate objects).
• Hands appear to be the major vector, and direct person-to-
person contact is the predominant mode of spread.
• These non-enveloped viruses are extremely stable and can
survive on such objects for many hours.

234
 Clinical Findings.
• Common cold
• The incubation period is brief, from 2 to 4 days, and the
acute illness usually lasts for 7 days although a non-
productive cough may persist for 2-3 weeks.
• The average adult has 1-2 attacks each year.
• Usual symptoms in adults include irritation in the upper
respiratory tract, nasal discharge, headache, mild cough,
malaise, and a chilly sensation. There is little or no fever.
• The nasal and nasopharyngeal mucosa become red and
swollen, and the sense of smell becomes less keen.

235
 Laboratory Diagnosis
• The clinical syndrome of the common cold is usually
so characteristic that laboratory diagnosis is
unnecessary.
• Virus can be obtained from nasal washings.
• Rhinoviruses are grown in human diploid fibroblast
cells (e.g., WI-38) at 33° C.
• genome analysis by RT-PCR.
• The performance of serologic testing to document
rhinovirus infection is not practical.

236
 Treatment, Prevention, and Control

• There are many over-the-counter remedies for the

common cold.
• Inhaling hot, humidified air, and even the steam from
hot chicken soup, may actually help by increasing
nasal drainage.
• Rhinovirus is not a good candidate for a vaccine
program.
• Handwashing and disinfection of contaminated
objects are the best means of preventing viral spread.

237
By Wondmagegn D
 Objectives

• Explain the properties of Hepatitis Virus

• Describe the pathogenesis and clinical
features Hepatitis virus infection
• Illustrate epidemiology Hepatitis Virus
• Describe the diagnosis Hepatitis Virus
infection

8/23/2020 239
 Outline
• Properties
• Classification and structure
• Pathogenesis
• Clinical features
• Diagnosis

8/23/2020 240
•
Introduction
The hepatitis alphabet of viruses includes at least six viruses,
A through G.

• Although the target organ for each of these viruses is the

liver and the basic hepatitis symptoms are similar, they differ
greatly in their structure, mode of replication, mode of
transmission, and in the time course and sequelae of the
disease they cause.

• Hepatitis A virus (HAV) and hepatitis B virus (HBV) are the

classic hepatitis viruses, and hepatitis C, G, E, and hepatitis D
virus (HDV), the delta agent, are called non-A, non-B hepatitis
(NANBH) viruses.
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• Each of the hepatitis viruses infects and damages
the liver, causing the classic icteric symptoms of
jaundice and the release of liver enzymes.

• The specific virus causing the disease can be

distinguished by the course, nature, and serology
of the disease.

• These viruses are readily spread because infected

people are contagious before, or even without,
showing symptoms.
 What makes hepatitis a global health problem?

• About 1 million deaths per year are attributed to viral

hepatitis infections.
• Together, hepatitis B virus and hepatitis C are the leading
cause of liver cancer in the world, accounting for 78
percent of cases.
• Nearly one out of every three people in the world has been
infected by HBV, and
• one in twelve live with chronic HBV or HCV infection.
• HBV/HIV and HCV/HIV coinfections are an increasing
problem in countries with HIV epidemics and among
injecting drug users.
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 Hepatitis A virus (HAV)

• Hepatitis A, which is sometimes known as infectious

hepatitis,
• (1) is caused by a picornavirus, (RNA) virus; genus-
Hepatovirus
• (2) is spread by the fecal-oral route;
• (3) has an incubation period of approximately 1 month,
after which icteric symptoms start abruptly;
• (4) does not cause chronic liver disease; and
• (5) rarely causes fatal disease.
• Only a single serotype and Seven genotypes
• Humans only reservoir
 Cont…
• The virus has icosahedral symmetry and non-
enveloped.
• Contain linear single :stranded RNA genome with a
size of 7.5 kb.
• Capsid is more stable to acid and heat than other
enteroviruses.
• Virus is relatively resistant to common disinfectants.
• Long lasting immunity follows infection by HAV.
• No carrier state exists.

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• It is the etiologic agent infectious hepatitis or
short incubation hepatitis.

• Estimated to be the cause of 40% of acute

hepatitis cases.

• There is no antigenic cross-reactivity with HBV or

with the other hepatitis viruses.

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 Cont…

Figure …. Structure of hepatitis A virus.

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 Epidemiology
• Causes approximately 40% of acute cases of hepatitis.
• The virus spreads readily in a community because
– most infected people are contagious 10 to 14 days before symptoms occur,
and
– 90% of infected children and 25% to 50% of infected adults have
inapparent but productive infections.
• The virus is spread via the fecal-oral route.
• Virus is spread in contaminated water, in food, and by
dirty hands.
• HAV is resistant to detergents, acid (pH of 1), and
temperatures as high as 60° C, and it can survive for
many months in fresh water and salt water.
 Distribution of HAV

High
High/intermediate
Intermediate
Low
Very low

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 Cont…

• Virus is abundant in the feces (with some

culturable from throat and saliva as well)
• Abrupt onset of symptoms (15 to 50 days
p.I.) and intensify 4 to 6 days before
icteric phase
• Clinical symptoms very similar to HBV
(malaise, lethargy) but may be less severe

8/23/2020 255
 Cont…
• Most infections (90%) occur in children who are
asymptomatic or an icteric (symptomatic without jaundice)

• Severity of the disease increases with age (50-75% of adult

infections are icteric)

• By the time “dark urine” appears, most of the virus is gone.

• Virus not cytopathic, liver damage due to cell mediated

immune response

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 Laboratory Diagnosis

• The diagnosis of HAV infection is generally made on the

basis of
– the time course of the clinical symptoms,
– identification of a known infected source, and
– most reliably, results of specific serologic tests.

• The best way to demonstrate an acute infection is by

finding anti-HAV IgM, as measured by ELISA or RIA.
• Virus isolation is not performed, because efficient tissue
culture systems for growing the virus are not available.
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 Treatment/Prevention
• Supportive- no specific role of antiviral therapy
• Lifelong immunity likely after infection only homologous
strains
• Interruption of fecal-oral spread
• Avoidance of contaminated water or food
(undercooked shell fish)
• Proper hand washing in day care and healthcare
facilities
• Prophylaxis with immune globulin before or early in
incubation (< 2wks post exposure) is 80 - 90% effective
• Killed vaccine is available for those at risk

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 Hepatitis B (HBV)

HBV
• HBV is a small enveloped DNA virus with several
unusual properties
• 1/3 of world population is infected (out of, ¼ end
up of complication)
• resulting in 1 to 2 million deaths per year.
• Hepatitis B virus (HBV) has been classified into 8
genotypes (A-H).
– Genotypes A, D and E predominate in Africa.
• It has not yet been possible to propagate the virus
in cell culture.
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• Hepatitis B, previously known as serum hepatitis,
• (1) is caused by a hepadnavirus with a deoxyribonucleic
acid (DNA) genome;
• (2) is spread parenterally by blood or needles, by sexual
contact, and perinatally;
• (3) has a median incubation period of approximately 3
months, after which icteric symptoms start insidiously;
• (4) is followed by chronic hepatitis in 5% to 10% of
patients; and
• (5) is causally associated with primary hepatocellular
carcinoma (PHCC).
• The incidence of HBV is decreasing, however, especially in
infants, because of the development and use of the HBV
subunit vaccine.
 structure
• Although a DNA virus, it encodes a reverse
transcriptase and replicates through an RNA
intermediate.
• The virion, also called the Dane particle, is 42 nm in
diameter.
• The virions are unusually stable for an enveloped
virus.
• They resist treatment with ether, low pH, freezing,
and moderate heating.
– assist transmission and hamper disinfection.
 Struc…
• The HBV virion includes a protein kinase and a
polymerase with reverse transcriptase and ribonuclease H
activity, as well as a P protein attached to the genome.
• All of this is surrounded by an icosahedral capsid formed
by the hepatitis B core antigen (HBcAg) and
• an envelope containing three forms of the glycoprotein
hepatitis B surface antigen (HBsAg).
• A hepatitis Be antigen (HBeAg) protein shares most of its
protein sequence with HBcAg
 Pathogenesis
• HBV can cause acute or chronic, symptomatic or
asymptomatic disease. Which of these occurs seems to be
determined by the person’s immune response to the
infection
• The major source of infectious virus is blood, but HBV can
be found in semen, saliva, milk, vaginal and menstrual
secretions, and amniotic fluid. The most efficient way to
acquire HBV is through injection of the virus into the
bloodstream
• Initial infection with HBV occurs through injection,
unprotected sex, and birth.
• The virus then spreads to the liver, replicates, induces a
viremia, and is transmitted in various body secretions in
addition to blood to start the cycle again.
• Symptoms are caused by
– cell-mediated immunity (CMI) and
– immune complexes between antibody and hepatitis B surface
antigen (HBsAg).
 Concentration of Hepatitis B Virus in Various
Body Fluids

Low/Not
High Moderate Detectable

blood semen urine

serum vaginal fluid feces
wound exudates saliva sweat
tears
breastmilk

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 Laboratory Diagnosis
• The initial diagnosis of hepatitis can be made on the basis of
the clinical symptoms and the presence of liver enzymes in
the blood.

• However, the serology of HBV infection describes the course

and nature of the disease.

• Acute and chronic HBV infections can be distinguished by

the presence of HBsAg and HBeAg in the serum and the
pattern of antibodies to the individual HBV antigens.
• HBsAg and HBeAg are secreted into the blood
during viral replication.

• The amount of virus in blood can be determined by

quantitative genome assays using polymerase chain
reaction (PCR) and related techniques.

• Knowing the viral load can help in following the

course of chronic HBV infection and antiviral drug
efficacy.
Treatment, Prevention, and Control

• Hepatitis B immune globulin may be administered

within a week of exposure and to newborn infants
of HBsAg positive mothers to prevent disease.

• Chronic HBV infection can be treated with drugs

targeted at the polymerase

• Pegylated interferon-α
• Transmission of HBV in blood or blood products
has been greatly reduced by
– screening donated blood for the presence of HBsAg and
anti-HBc.

• Additional efforts to prevent transmission of HBV

include safe sex and avoiding lifestyles that
facilitate spread of the virus.

• Vaccination is recommended for infants, children,

and especially people in high-risk groups
• at increased risk are,
– Household contacts,
– sexual partners of HBV carriers,
– patients undergoing hemodialysis,
– recipients of pooled plasma products,
– health care workers exposed to blood,
and
– babies born to HBV-carrier mothers.
CONTROL
 HDV
• Approximately 15 million people in the world are infected
with HDV (delta agent), and
• the virus is responsible for causing 40% of fulminant
hepatitis infections.
• is unique in that it requires actively replicating HBV as a
“helper virus” and
• occurs only in patients who have active HBV infection.
• HBV provides an envelope for HDV RNA and its antigens.
• HDV exacerbates the symptoms caused by HBV.
 Hepatitis C (HCV)

General Features
• HCV is the only member of the Hepacivirus genus
of the Flaviviridae family.

• Enveloped, (+) ssRNA genome, icosahedral capsid

• Originally referred to as “non-A, non-B hepatitis

(NANB)”

• Humans and chimpanzees only known reservoirs

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• There are six major genotypes of HCV (clades),
– between and within each genotype there is considerable
genetic and antigenic diversity.
• The genome of HCV (9100 nucleotides) encodes 10
proteins, including two glycoproteins (E1, E2).
• The viral RNA-dependent RNA polymerase is error
prone
– generates mutations in the glycoprotein and other genes.
• This generates antigenic variability….makes
development of a vaccine very difficult.
 Clinical feature
Disease
• Spread via infected blood and sexual contact- Target
organ liver
• 6 - 8 week incubation period
• most infections are sub-clinical
• Clinical infections are generally less severe than HBV,
damage due to cell mediated immune response.
• HCV has a higher incidence of chronic liver disease than
HBV (70% of patients remain viremic for more than 1
year)

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• HCV causes three types of disease:
– acute hepatitis with resolution of the infection
and recovery in 15% of cases,
– chronic persistent infection with possible
progression to disease much later in life for
70% of infected persons, and
– severe rapid progression to cirrhosis in 15% of
patients.
 Epidemiology of HCV
• Hepatitis C virus (HCV) is a major cause of liver
disease worldwide and a potential cause of
substantial morbidity and mortality in the future.
• 3% world population
• High—Africa, South America, Asia
• 170 million chronic carriers
• Transmission—pre-cutaneous exposure to blood
• Drug users, sex, health practitioners, mother to
infants.

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 Chronic Hepatitis C
Factors Promoting Progression or Severity

• Increased alcohol intake

• Age > 40 years at time of infection

• HIV co-infection

• Chronic HBV co-infection

8/23/2020 289
 Laboratory diagnosis
• ELISA recognition of anti-HCV antibody or detection of
the RNA genome.
– Seroconversion occurs within 7 to 31 weeks of infection.
– used for screening the blood supply from normal donors.

• Antibody is not always detectable in viremic people,

immunocompromised patients, or those receiving
hemodialysis.
• Genome detection and quantitation by RT-PCR, branched-
chain DNA, and related techniques
– is the gold standard for confirming a diagnosis of HCV and for
following the success of antiviral drug therapy.

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 Treatment/Prevention
• No vaccine and no post-exposure prophylaxis for HCV,
• the focus of primary prevention efforts should be;
– safer blood supply in the developing world,
• Screening and testing blood, plasma, organ, tissue and semen
donors
– safe injection practices in health care and other
settings, and
– decreasing the number of people who initiate
injection drug use.
• Counseling drug users and safe sexual practice
• Alpha-interferon is the only reliable treatment and only
moderately successful- serotype specific

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 Hepatitis E (HEV)

General Features
• HEV (E-NANBH) is predominantly spread by the fecal-
oral route, especially in contaminated water.
• HEV is unique but resembles the caliciviruses, based on
its size (27 to 34 nm), RNA genome, and naked capsid
structure.
• Although HEV is found throughout the world, it is most
problematic in developing countries.

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• The symptoms and course of HEV disease are
similar to those of HAV disease;
• it causes only acute disease.
• However, the symptoms for HEV may occur later
than those of HAV disease.
• The mortality rate associated with HEV disease is
1% to 2%, approximately 10 times that associated
with HAV disease.
• HEV infection is especially serious in pregnant
women (mortality rate of ≈20%).
 Cont…

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 Hepatitis G (HGV)

General Features
• Flavivirus
• Parenteral transmission (esp. i.v. drug use)
• Sexual transmission?
• Newly characterized NANBH
• Role ???

8/23/2020 295
 Pathogenesis

• Estimated to cause 0.3% of acute viral hepatitis

• 900 - 2000 infections per year, mostly
asymptomatic
• Chronic disease? Controversial
• Diagnosis RNA based methods

8/23/2020 296
 Cont…

Risk groups
• Transfusion recipients
• Injection drug users
• Frequent co-infection with hepatitis C

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 Viral Hepatitis - Overview
Type of Hepatitis
A B C D E

Source of feces blood/ blood/ blood/ feces

virus blood-derived blood-derived blood-derived
body fluids body fluids body fluids

Route of fecal-oral percutaneous percutaneous percutaneous fecal-oral

transmission permucosal permucosal permucosal

Chronic no yes yes yes no

infection

Prevention pre/post- pre/post- blood donor pre/post- ensure safe

exposure exposure screening; exposure drinking
immunization immunization risk behavior immunization; water
modification risk behavior
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Rhabdoviruses and Filoviruses
 Introduction
• The members of the family Rhabdoviridae include
pathogens for a variety of mammals, fish, birds, and
plants.

• The family contains

– Vesiculovirus (vesicular stomatitis viruses[VSVs]),
– Lyssavirus (rabies and rabies-like viruses),
– unnamed genus constituting the plant rhabdovirus
group, and
– ungrouped rhabdoviruses of mammals, birds, fish ...
• Rabies virus is the most significant pathogen

8/23/2020 300
 The rabies virus
• Are simple viruses encoding only five proteins and
appearing as bullet-shaped, enveloped virions

• The helical nucleocapsid, is composed of

• one molecule of SS (-)RNA and the
nucleoprotein (N), large (L), and nonstructural
(NS) proteins.
• Prototype for replication of negative-strand
enveloped viruses

8/23/2020 301
 The rabies virus
• The matrix (M) protein lies between the envelope and the
nucleocapsid.

• The N protein is the major structural protein of the virus.

It protects the RNA from ribonuclease digestion.

• The L and NS proteins constitute the RNA-dependent

RNA polymerase.

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• Until Louis Pasteur developed the killed –rabies
vaccine, a bite from a “mad dog” always led to the
characteristic symptoms of hydrophobia and certain
death.

• In 1944 Negri made the first histological diagnosis of

rabies when he observed inclusion bodies in the
cytoplasm of nervous tissue.

• The inclusion bodies bear his name Negir bodies.

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8/23/2020 304
 Disease Mechanisms of Rabies Virus
• Rabies is usually transmitted in saliva and is
acquired from the bite of a rabid animal.

• Virus replicates in the muscle at the site of the bite,

with minimal or no symptoms (incubation phase).

• The length of the incubation phase is determined

by the infectious dose and the proximity of the
infection site to the central nervous system (CNS)
and brain.
8/23/2020 305
 Disease Mechanisms of Rabies Virus
• After weeks to months, the virus infects the
peripheral nerves and travels up the CNS to the
brain (prodrome phase).

• Infection of the brain causes classic symptoms,

coma, and death (neurologic phase).

• During the neurologic phase, the virus spreads

to the glands, skin, and other body parts,
including the salivary glands, from where it is
transmitted.
8/23/2020 306
• Rabies infection does not elicit an antibody
response until the late stages of the disease.

• Administration of antibody can block the

progression of the virus and disease if given early
enough.

• The long incubation period allows active

immunization as a post exposure treatment.

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 Pathogenesis of Rabies
Virus

8/23/2020 308
 Epidemiology of Rabies Virus
• Rabies is the classic zoonotic infection spread from
animals to humans .

• It is endemic in a variety of animals worldwide,

except in Australia.

• Rabies is maintained and spread in two ways.

I. In urban rabies, dogs are the primary transmitter, and
II. in sylvatic (forest) rabies, many species of wildlife can
serve as transmitters
• Disease has long, asymptomatic incubation period
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8/23/2020 312
 Clinical Syndromes
• Acute viral disease of CNS
• Rabies is virtually always fatal unless treated by
vaccination.

1. Incubation Phase----long but highly variable

2. Prodrome phase----The patient has symptoms such

as fever, malaise, headache, pain or paresthesia
(itching) at the site of the bite, gastrointestinal
symptoms, fatigue, and anorexia. lasts 2 to 10 days,
8/23/2020 314
 Clinical Syndromes
3. Neurologic phase----The neurologic symptoms
specific to rabies appear. Hydrophobia, seizures,
disorientation, and hallucinations . The paralysis may
lead to respiratory failure.

4. The patient becomes comatose after the neurologic

phase, which lasts from 2 to 10 days. This phase
almost universally leads to death resulting from
neurologic and pulmonary complications.

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 Time course of Rabies infections

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 Laboratory Diagnosis
• Laboratory tests are usually performed to confirm
the diagnosis and to determine whether a suspected
individual or animal is rabid (postmortem).
Isolation of virus (saliva, CSF, brain )
Serology
Viral Ag detection ( infected tissue )
Viral RNA detection ( PCR )

8/23/2020 317
Diagnosis: Rabies virus Negri bodies

H&E staining of neural tissue

CDC Website: http://www.cdc.gov/ncidod/dvrd/rabies/diagnosis/diagnosi.htm

8/23/2020 318
 Treatment and Prophylaxis
• Clinical rabies is almost always fatal unless treated with post
rabies immunization.

• Post exposure prophylaxis is the only hope for preventing

overt clinical illness in the affected person.

• The first protective measure is local treatment of the wound.

• The wound should be washed immediately with soap and

water or another substance that inactivates the virus.

8/23/2020 319
 Treatment and Prophylaxis
• killed-virus vaccine: chemical inactivation of rabies infected–
tissue culture human diploid cells (HDCV).
• Preexposure vaccination(animal workers, laboratory workers
and people traveling to areas where rabies is endemic.

• HDCV administered intramuscularly or intradermally in three

doses is recommended and provides 2 years of protection.

• Ultimately the prevention of human rabies hinges on the

effective control of rabies in domestic and wild animals.

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 Filoviruses
• The Marburg and Ebola viruses were classified as
members of the family Rhabdoviridae but are now
classified as filoviruses (Filoviridae).

• They are filamentous, enveloped, negative-strand

RNA viruses.

• These agents cause severe or fatal hemorrhagic

fevers and are endemic in Africa.
 Structure and Replication
• Filoviruses have a single-stranded RNA
genome that encodes seven proteins.

• The virions form enveloped filaments

• The nucleocapsid is helical and enclosed in

an envelope containing one glycoprotein.
 Pathogenesis
• The filoviruses replicate efficiently, producing large
amounts of virus in endothelial cells, monocytes,
macrophage, dendritic cells, and other cells.
• Replication in monocytes elicits a cytokine storm of
proinflammatory cytokines
• Viral cytopathogenesis causes extensive tissue necrosis in
parenchymal cells of the liver, spleen, lymph nodes, and
lungs.
• Infection of endothelial cells leading to vascular injury and
leakage.
• The widespread hemorrhage that occurs in affected
patients causes edema and hypovolemic shock.
 Epidemiology
• These viruses may be endemic in bats or wild monkeys
and can be spread to humans and between humans.

• Contact with the animal reservoir or direct contact with

infected blood or secretions can spread the disease.

• These viruses have been transmitted by accidental injection

and through the use of contaminated syringes.
 Clinical Syndromes
• Marburg and Ebola viruses are the most severe causes of
viral hemorrhagic fevers.
• The illness usually begins with flulike symptoms such as
headache and myalgia.
• Nausea, vomiting, and diarrhea occur within a few days; a
rash also may develop.
• Subsequently, hemorrhage from multiple sites (especially
the gastrointestinal tract) and
• death occur in as many as 90% of patients with clinically
evident disease.
 Laboratory Diagnosis
• All specimens from patients with a suspected filovirus
infection must be handled with extreme care to prevent
accidental infection.
• Handling of these viruses requires level 4 isolation
procedures that are not routinely available.
• Viral antigens can be detected in tissue by direct
immunofluorescence analysis and in fluids by ELISA.
• RT-PCR amplification of the viral genome in secretions
can be used to confirm the diagnosis and minimize
handling of samples.
 Treatment, Prevention, and Control

• Antibody-containing serum, artificially produced antibody

and interferon and ribavirin therapies have been tried in
patients with filovirus infections.

• Infected patients should be quarantined, and contaminated

animals should be sacrificed.

• Handling of the viruses, infected individuals, dead bodies,

and contaminated materials requires very stringent
isolation procedures.
Reoviridae
• The Reoviridae consist of the orthoreoviruses, rotaviruses,
orbiviruses, and coltiviruses.
• The name was proposed for a group of respiratory and enteric
viruses that were not associated with any known disease
(respiratory, enteric, orphan).
• The Reoviridae are nonenveloped viruses with double-layered
protein capsids containing 10 to 12 segments of the dsRNA.
• These viruses are stable in detergents, over wide pH and
temperature ranges, and in airborne aerosols.
• The orbiviruses and coltiviruses are spread by arthropods and
are arboviruses.

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• Reoviruses are double-stranded, segmented RNA genome.

• The family includes human rotaviruses, the most

important cause of infantile gastroenteritis.

• In developing countries it is estimated to cause as many

as 1.5 million deaths of preschool children annually, of
which rotavirus is responsible for about 600,000 deaths.

• In the United States, acute gastroenteritis is second only

to acute respiratory infections as a cause of disease in
families.

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 Classification
• The family Reoviridae is divided into twelve genera. Four of
the genera are able to infect humans and animals:
Orthoreovirus, Rotavirus, Coltivirus, and Orbivirus.

• There are at least five species or groups of rotaviruses (A–E),

plus two tentative species (F and G), of which three species
(A, B, C) infect humans.

• Strains of human and animal origin may fall in the same

serotype.

• Other rotavirus groups and serotypes are found only in

animals.

• 3 serotypes of reovirus, 100 different orbivirus serotypes and

two coltivirus serotypes.
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 Rotaviruses

• Rotaviruses are a major cause of diarrheal illness in

human infants and young animals, including calves
and piglets.

• Infections in adult humans and animals are also

common.

• Rotaviruses resemble reoviruses in terms of

morphology and strategy of replication.

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 Animal Susceptibility

• Rotaviruses have a wide host range. Most isolates

have been recovered from newborn animals with
diarrhea.
• Cross-species infections can occur in experimental
inoculations, but it is not clear if they occur in
nature.
• Newborns often exhibit subclinical infection due
perhaps to the presence of maternal antibody.

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 Clinical Findings
• Rotaviruses cause the major portion of diarrheal illness in
infants and children worldwide but not in adults .
• There is an incubation period of 1–3 days.
• Typical symptoms include watery diarrhea, fever,
abdominal pain, and vomiting, leading to dehydration.
• In infants and children, severe loss of electrolytes and
fluids may be fatal unless treated.
• Patients with milder cases have symptoms for 3–8 days
and then recover completely.
• viral excretion in the stool may persist up to 50 days after
onset of diarrhea
• In children with immunodeficiency's, rotavirus can cause
severe and prolonged disease.
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 Cont…

• Adult contacts may be infected, as evidenced by

seroconversion, but they rarely exhibit symptoms,
and virus is infrequently detected in their stools.

• A common source of infection is contact with

pediatric cases.

• Epidemics of severe disease have occurred in adults,

especially in closed populations, as in a geriatric
ward.

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 Epidemiology & Immunity
• Rotaviruses are the single most important worldwide
cause of gastroenteritis in young children.
• Estimates range from 3 billion to 5 billion for annual
diarrheal episodes in children under 5 years of age
and as many as 1 million deaths.
• Developed countries --high morbidity but a low
mortality.
• Typically, up to 50% of cases of acute gastroenteritis
of hospitalized children throughout the world are
caused by rotaviruses. Nosocomial infections are
frequent.

• Transmission appears to be by the fecal-oral route.

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 Cont…

• By age 3 years, 90% of children have serum antibodies to

one or more types.
• Rotavirus reinfections are common; it has been shown
that young children can suffer up to five reinfections by
2 years of age.
• Asymptomatic infections are more common with
successive reinfections.
• Local immune factors, such as secretory IgA or
interferon, may be important in protection against
rotavirus infection.
• Asymptomatic infections are common in infants before
age 6 months, the time during which protective maternal
antibody acquired passively by newborns should be
present.
• Such neonatal infection does not prevent reinfection, but
it does protect against the development of severe
disease during reinfection
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 Laboratory Dx

• Laboratory diagnosis rests on demonstration of virus

in stool collected early in the illness and on a rise in
antibody titer.
• Virus in stool is demonstrated by IEM, latex
agglutination tests, or ELISA.
• Genotyping of rotavirus nucleic acid from stool
specimens by the polymerase chain reaction is the
most sensitive detection method.
• Serologic tests can be used to detect an antibody
titer rise, particularly ELISA.
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 Treatment & Control
• Treatment of gastroenteritis is supportive, to correct
the loss of water and electrolytes that may lead to
dehydration, acidosis, shock, and death.
• Management consists of replacement of fluids and
restoration of electrolyte balance either intravenously or
orally, as feasible.
• The infrequent mortality from infantile diarrhea in
developed countries is due to routine use of effective
replacement therapy.
• In view of the fecal-oral route of transmission,
wastewater treatment and sanitation are significant
control measures.

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 Treatment & Control

• A safe and effective vaccine remains the best hope for

reducing the worldwide burden of rotavirus disease.

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Togaviridae and
Flaviviridae
 Objectives

• Explain the properties of Toga and Fiavivirus

• Describe the pathogenesis and clinical features Toga
and flavivirus infection.
• Illustrate epidemiology Toga and Flavivirus infection
• Describe the diagnosis Toga and Flavivirus infection
• Describe the prevention and control of Toga and
Flavivirus infection

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 Outline
• Introduction
• Classification and structure
• Pathogenesis
• Clinical features
• Diagnosis

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 Introduction
• The members of the Togaviridae and Flaviviridae families
are enveloped, positive, single-stranded ribonucleic acid
(RNA) viruses
• Alphavirus and Flavivirus
– share similarities in the diseases that they cause and
in their epidemiology.
– Most are transmitted by arthropods and are
therefore arboviruses (arthropod-borne viruses).
– They differ in size, morphology, gene sequence, and
replication.

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 Togaviridae
1. Alphavirus
a) Sindbis
b) Semliki Forest
c) Venezuelan equine encephalitis
d) Eastern equine encephalitis
e) Western equine encephalitis
f) Chikungunya
2. Rubivirus
a) Rubella virus (German Measles)

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 Flaviviridae
1. Flavivirus includes mostly arthropod-borne viruses
a) Dengue fever
b) Yellow fever
c) Japanese encephalitis
d) West Nile encephalitis
e) St. Louis encephalitis
f) Russian spring-summer encephalitis
g) Powassan encephalitis
2. Hepaciviruses
a) Hepatitis C & G virus

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 ALPHAVIRUSES AND FLAVIVIRUSES
• The alphaviruses and flaviviruses are classified as
arboviruses because they are usually spread by
arthropod vectors.

• These viruses have a very broad host range,

including vertebrates (e.g., mammals, birds,
amphibians, reptiles) and invertebrates (e.g.,
mosquitoes, ticks).

• Diseases spread by animals or with an animal

reservoir are called zoonoses.
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 Cont…
• Togavirus replication includes early (nonstructural)
and late (structural) protein synthesis.

• Togaviruses replicate in the cytoplasm and bud at

the plasma membranes.

• Flaviviruses replicate in the cytoplasm and bud at

intracellular membranes.

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 Replication
1. Virion RNA acts as mRNA and is translated into
a polyprotein which is co-translationally cleaved
(post-translational cleavage for Togaviridae) to
yield non-structural (RNA dependent RNA
polymerase) and structural proteins.
2. Cytoplasmic site of replication
3. Genes for the structural and non-structural
proteins are located differently in the two virus
classes.
4. Buds through plasma membrane (togavirus) or
ER or Golgi (flavivirus)
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Arboviruses: Vectors, Hosts, and Diseases

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 Patterns of Alphavirus and Flavivirus Transmission

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 Disease Mechanisms of Togaviruses and Flaviviruses

• Viruses are cytolytic, except for rubella and hepatitis C.

• Viruses establish viremia and systemic infection.
• Viruses are good inducers of interferon, which can
account for the flulike symptoms during prodrome.
• Viruses, except rubella and hepatitis C, are arboviruses.
• Flaviviruses can infect cells of the monocyte-
macrophage lineage.
• These viruses are associated with mild systemic disease,
encephalitis, arthrogenic disease, or hemorrhagic
disease.

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 Cont…
• The initial viremia produces systemic symptoms, such as
fever, chills, headaches, backaches, and other flulike
symptoms, within 3 to 7 days of infection.
• Some of these symptoms can be attributed to the
effects of the interferon produced in response to the
viremia and Infection of host cells.
• The viremia is considered a mild systemic disease, and
most viral infections do not progress beyond this point.
• A secondary viremia can produce sufficient virus to infect
target organs, such as the brain, liver, skin, and
vasculature, depending on the tissue tropism of the virus

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 Epidemiology of Alphavirus and Flavivirus Infection

• Disease/Viral Factors
– can be inactivated by drying, soap, and detergents
– Virus can infect mammals, birds, reptiles, and insects
– Asymptomatic or nonspecific (flulike fever or chills),
encephalitis, hemorrhagic fever, or arthritis
• Transmission
– Specific arthropods characteristic of each virus
(zoonosis: arbovirus)
• Who Is at Risk?
– People who enter ecologic niche of arthropods

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 Cont…
• Geography/Season
– Endemic regions for each arbovirus are determined
by habitat of mosquito or other vector
– Aedes mosquito, which carries dengue and yellow
fever, is found in urban areas and in pools of water
• Modes of Control
– Mosquito breeding sites and mosquitoes should be
eliminated
– Live attenuated yellow fever virus and inactivated
Japanese encephalitis virus vaccines

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 Cont…
• Most flavivirus infections are relatively benign, but
serious aseptic meningitis and encephalitic or
hemorrhagic disease can occur.

• The encephalitis viruses include St. Louis, West Nile,

Japanese, Murray Valley, and Russian spring-
summer viruses.

• The hemorrhagic viruses are dengue and yellow

fever viruses.
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• Dengue virus is a major worldwide problem, with
up to 100 million cases of dengue fever and
300,000 cases of dengue hemorrhagic fever
(DHF) occurring per year.

• Dengue fever is also known as break-bone fever;

the symptoms and signs consist of high fever,
headache, rash, and back and bone pain that last
6 to 7 days

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 Cont…
• Yellow fever infections are characterized by severe
systemic disease, with degeneration of the liver,
kidney, and heart, as well as hemorrhage.

• Liver involvement causes the jaundice from which

the disease gets its name, but massive
gastrointestinal hemorrhages (“black vomit”) may
also occur.

• The mortality rate associated with yellow fever

during epidemics is as high as 50%.
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 Laboratory Diagnosis
• Cell Culture: The alphaviruses and flaviviruses can be
grown in both vertebrate and mosquito cell lines,
but most are difficult to isolate.

• Infection can be detected through the use of

cytopathologic studies, immunofluorescence, and
the hemadsorption of avian erythrocytes.

• Detection and characterization can be performed by

RT-PCR testing of genomic RNA or viral mRNA in
blood or other samples.
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Treatment, Prevention, and Control
• No treatments exist for arbovirus diseases,
other than supportive care.

• The easiest means of preventing the

spread of any arbovirus is elimination of its
vector and breeding grounds.

• A live vaccine against yellow fever virus

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• በድሬዳዋ ቺኩን ጉንያ የተባለ አዲስ ወረርሽኝ
ተከሰተ
• ድሬዳዋ — ቺኩን ጉንያ የተባለው በሽታ፣ ከዚህ
በፊት በአፋርና ሶማሌ ክልልም ታይቶ ነበር
ተብሏል፡፡ እስካሁን ከበሽታው ጋር ተመሳሳይ
ምልክት የታየባቸው 3756 ህሙማን
መገኘታቸውም ተገልጿል፡፡

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• Dire Dawa Health bureau reporting 3,756 cases with
similar symptoms of the Chikungunya virus in the
administration.

• In a statement today the bureau said it is working to

prevent the outbreak of Chikungunya in Dire Dawa.

• Since the disease has similar symptoms with malaria

and Dengue fever, the bureau did not confirm
whether all the patients are affected with the
Chikungunya virus.

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 Symptoms

• Symptoms usually begin 3–7 days after being

bitten by an infected mosquito.
• The most common symptoms are fever and joint
pain.
• Other symptoms may include headache, muscle
pain, joint swelling, or rash.
• Chikungunya disease does not often result in
death, but the symptoms can be severe and
disabling.
• Once a person has been infected, he or she is
likely to be protected from future infections.

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 Treatment
• There is no vaccine to prevent or medicine to treat
chikungunya virus.
• Treat the symptoms:
• Get plenty of rest.
• Drink fluids to prevent dehydration.
• Take medicine such as acetaminophen (Tylenol®) or
paracetamol to reduce fever and pain.

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 Rubella
• Rubella virus has the same structural properties and
mode of replication as the other togaviruses.

• However, unlike the other togaviruses, rubella is a

respiratory virus and does not cause readily
detectable cytopathologic effects.

• Rubella is one of the five classic childhood

exanthems, along with measles, roseola, fifth
disease, and chickenpox.
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 Pathogenesis
• Rubella infects the URT and then spreads to local lymph
nodes, which coincides with a period of lymphadenopathy.

• This stage is followed by establishment of viremia, which

spreads the virus throughout the body. Infection of other
tissues and the characteristic mild rash occur.
• The prodromal period lasts approximately 2 weeks .

• The infected person can shed virus in respiratory droplets

during the prodromal period and for as long as 2 weeks after
the onset of the rash.

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 Immune Response

• Antibody is generated after the viremia, and its appearance

correlates with the appearance of the rash.
• The antibody limits viremic spread, but cell-mediated immunity
plays an important role in resolving the infection.
• Only one serotype of rubella exists, and natural infection produces
lifelong protective immunity
• Most important, serum antibody in a pregnant woman prevents
spread of the virus to the fetus.
• Immune complexes most likely cause the rash and arthralgia
associated with rubella infection.

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 Congenital Infection
• Rubella infection in a pregnant woman can result
in serious congenital abnormalities in the child.

• If the mother does not have antibody, the virus

can replicate in the placenta and spread to the
fetal blood supply and throughout the fetus.

• Rubella can replicate in most tissues of the fetus.

The virus may not be cytolytic, but the normal
growth, mitosis, and chromosomal structure of
the cells of the fetus can be altered by the
infection.
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 Epidemiology of Rubella Virus
• Disease/Viral Factors
• Rubella infects only humans
• Virus can cause asymptomatic disease
• There is one serotype
• Transmission
• Respiratory route
• Who Is at Risk?
• Children: mild exanthematous disease
• Adults: more severe disease with arthritis or arthralgia
• Neonates younger than 20 weeks: congenital defects
• Modes of Control
• Live attenuated vaccine administered as part of the
MMR vaccine
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 Clinical Syndromes
• Rubella disease is normally benign in children. After a 14-
to 21-day incubation period, the symptoms in children
consist of a 3-day maculopapular or macular rash and
swollen glands.

• Infection in adults, however, can be more severe and

include problems such as bone and joint pain (arthralgia
and arthritis) and (rarely) thrombocytopenia or post
infectious encephalopathy.

• Congenital disease is the most serious outcome of rubella

infection. The fetus is at major risk until the 20th week
of pregnancy.
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• Maternal immunity to the virus resulting from
prior exposure or vaccination prevents spread of
the virus to the fetus.
• The most common manifestations of congenital
rubella infection are
• cataracts,
• mental retardation,
• cardiac abnormalities,
• Intrauterine growth retardation and
• deafness
• The mortality in utero and within the first year
after birth is high for affected babies.
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 Laboratory Diagnosis
• Isolation of the rubella virus is difficult and rarely
attempted.
• The presence of the virus can be detected by RT-PCR
detection of viral RNA.
• The diagnosis is usually confirmed by the presence of anti
rubella-specific IgM.
• A fourfold increase in specific IgG antibody titer between
acute and convalescent sera is also used to indicate a
recent infection.
• Antibodies to rubella are assayed early in pregnancy to
determine the immune status of the woman.

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 Reading assignment

BUNYAVIRIDAE AND ARENAVIRIDAE

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