ORTHOMYXOVIRUSES

(Influenza virus A, B and C)

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BY

D R EI LU

BMS 3.1CLINICAL VIROLOGY. SEPT, 2022

Orthomyxoviruses (Influenza viruses)
Name is from the Greek 'myxa' = mucus!
‘Orthos’- means “straight”
Belongs to the family Orthomyxoviridae.
Most important virus in this family is the
Influenza Virus.
The virus was first isolated in 1933 in
ferrets(kind of wild cats)
There are three antigenic types of influenza
viruses; A,B & C.
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 Introduction cont.
Influenza virus
 Influenza A- pandemics and epidemics;
humans,animals and birds
 Influenza B- epidemics; human
virus(almost an exclusive human
pathogen).
 Influenza C- mild respiratory tract
infection( infects humans and also pigs).

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 Properties
Linear segmented (8 segments) (-) ssRNA
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Helical capsid (spirally-shaped protein coat)

Enveloped
 Envelope contain 2 spikes
 Hemagglutinin
Binds to cell surface receptors( neuraminic acid/sialic
acid
 Neuraminidase
Enzymatic activity

There are 4 antigens present, the haemagglutinin (HA), neuraminidase

(NA), matrix (M-protein) and the nucleocapsid proteins.
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 The NP is a type-specific antigen which occurs in 3

forms, A, B and C, which provides the basis for the
classification of human influenza viruses

 The matrix protein (M protein) surrounds the

nucleocapsid and makes up 35-45% of the particle
mass.
Structure

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 Structure
Shape: pleomorphic, usually spherical
about 80-120 nm in diameter, enveloped.
Genome: single-stranded, RNA, negative
sense, segmented (8 segments in types A & B,
7 segments in type C lacking the
neuraminidase gene).
Envelope: derived from the cell, lipid, two
glycoprotein spikes project from its outer
surface: haemagglutinin (HA) &
neuraminidase (NA).
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 Classification
There are three antigenic types A, B & C,
classified according to the NP & M1
proteins.
Type A is divided into subtypes according
to HA & NA.
There are 15 types of HA & 9 types of NA
in different combinations. Three HA (H1-
H2, H3) & two NA (N1, N2) subtypes have
been recovered from humans.
Influenza A subtypes & B subtypes are
further divided into strains.

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 Antigenic variation
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Antigen variation of hemaglutinin and

neuroaminidase is responsible for epidemics and
pandemics

2 types of antigenic variation are known; antigenic

shift and antigenic drift.
Antigenic drift
 Point mutations in the genes specifying the
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surface proteins of a virus
 Sudden Change in the amino acid
sequence of the H Ag
 Step-by-step change whereby surface antigens of
haemagglutination (H) and neuraminidases (N)
change slightly or more considerably over a period
of time
 Occur both in A & B
 Antigenic shift
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Sudden appearance of a virus with widely divergent

antigenic structure
 Undergoes reassortment
 Results in changes of the H and N antigen
 Pandemics and epidemics
 Occurs with influenza A only

Usually occurs in one geographical region

There is genetic reassortment between two subtypes of a

viral species resulting in emergency of a new subtype with a
completely different surface protein
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Common among viruses with segmented

genomes

 Epidemics due to new virus strains arising due to

antigenic drift is not as great as for those showing
antigenic shift, since partial immunity is present in
persons with cross- reacting antibody induced by
previous infection.
Insert figure 25.3
Antigenic shift

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 Pathogenesis of Influenza viruses.
Spread:
 Person to person spread is primarily via aerosols.
 Incubation period is: 18-72 hrs.
 Virus concn in nasal & tracheal secretions is high for 24-48
hrs after contact, but may be take longer in children.
Site of infection:
 Influenza virus infects resp. tract epithelial cells.
 The ciliary cells and mucus producing cells are affected
 Efficiency of ciliary clearance is reduced & reduced clearance
of infectious agents.
 The symptoms of influenza are caused by viral pathologic
and immunopathologic effects due to over production of IFN
and also cytotoxic T-cells
 Viremia is very rare
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 Pathogenesis
When influenza virus is introduced into the respiratory tract, by aerosol or
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by contact with saliva or other respiratory secretions from an infected
individual
The virus particles binds to cells of the respiratory epithelium which are
rich in viral receptors.
Neuraminidase present on the virus particles aid the infectious process by
releasing virus particles which have been bound by the mucous present on
the surface of epithelial cells
The virus replicates in cells of both the upper and lower respiratory tract.
In primary viral pneumonia, the virus replicates in alveolar epithelial cells,
leading to rupture of walls of alveoli and bronchioles.
Viral replication combined with the immune response to infection lead to
destruction and loss of cells lining the respiratory tract.
Influenza H5N1 viruses frequently cause primary viral pneumonia
characterized by diffuse alveolar damage and interstitial fibrosis.
As infection subsides, the epithelium is regenerated, a process that can take
up to a month. Cough and weakness may persist for up to 2 weeks after
infection.
Recovery: Pathogenesis cont.
IFN may play a role by reducing virus
replication.
Many of the symptoms of uncomplicated
influenza (muscle aches, fatigue, fever) are of
IFN.
The cell-mediated immune response is
important in viral clearance.
The Ab response is usually not significant
until after virus has been cleared.
Repair of the resp. epithelium begins
rapidly, but may take some time to complete.
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 Vertical transmission:
The H5N1 bird flu virus can also pass through a
pregnant woman's placenta to infect the
fetus→affects the lungs, but also passes
throughout the body into the gastrointestinal
tract, the brain, liver, and blood cells.

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 Clinical features
Following a typical incubation period of 48 hours, the
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typical symptoms of influenza appears

 The onset is abrupt with a marked fever, headache,

photophobia, shivering, a dry cough, malaise, myalgia,
and a dry tickling throat, muscle aches, conjunctivitis

The fever is continuous and lasts around 3 days

Influenza B infection is similar to influenza A

 Infection with influenza C is usually subclinical or very

mild in nature
Complications
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1. Tracheobronchitis, bronchiolitis and croup
2. Pneumonia
 Secondary bacterial pneumonia - usually occurs
late in the course of disease, after a period of
improvement has been observed for the acute disease.

 S. aureus is most commonly involved although S.

pneumoniae and H. influenzae may be found.
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Pneumonia; inflammation of the air sacs (alveoli) in

one or both lungs, which may fill air sacs with fluid or pus
(purulent material)
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3. Myositis and myoglobinuria

4. Reye's syndrome - Reye's syndrome is characterized
by encephalopathy and fatty liver degeneration (swelling
in brain & Liver)

5. Other complications - influenza infection have been

implicated in acute viral encephalitis and Guillain-Barre
syndrome

-Influenza A was also associated with the cot death

syndrome.
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 Laboratory diagnosis
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1. Rapid Diagnosis - Nasopharyngeal aspirates are

the specimens of choice

Throat and nasal swabs are more commonly used

given the difficulties involved in taking
nasopharyngeal aspirates

Cells from pathological specimens may be examined

for the presence of influenza A and B antigens by
indirect immunofluorescence.
EIA tests for the detection of influenza A viral
antigens are available that24are easier but are less
sensitive

RT-PCR assays are available for the detection of

influenza RNA

Very sensitive and specific. Used in detection of

2009 pandemic H1N1 virus

RT-PCR assays are very expensive

Virus Isolation - Throat swabs, NPA and nasal
washings are used for virus isolation
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Nasal washings are the best specimens for virus

isolation

The specimen may be inoculated in embryonated

eggs or tissue culture

Pathological specimens can be inoculated on to tissue

cultures of kidney, chicks or a variety of other species.
Rhesus monkey cells are the most sensitive.
Newly produced virus can26be recognized by
haemadsorption and haemagglutination

 Influenza B virus and occasionally influenza A will

produce a CPE in MDCK cells

 Influenza viruses isolated from embryonated eggs or

tissue culture can be identified by serological or
molecular methods
Influenza viruses can be recognized as A, B, or C
types by the use of complement fixation tests (CFT)
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against the soluble antigen

The HA type is identified by HAI tests, the NA type

is also identified
3. Serology - Virus cannot be isolated from all cases
of suspected infection
Arising serum antibody to the infecting virus is
detected
 CFT is the most common method used using the
type specific soluble antigen
However, the CF test is thought to have a low
specificity
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A more specific test is the HAI test, but may not

accurately detect smaller increases in antibody

 A more precise method for measuring antibody is by

SRH.(single Rapid Hemolysis) SRH is more sensitive
than CF or HAI tests and has a greater degree of
precision

 A 50% increase in zone area represents a rise in

antibody and is evidence of recent infection.
 Treatment
Treatment of influenza is 29
entirely symptomatic

Salicylates should be avoided in children because of

the link with Reye's syndrome

 Neuraminidase inhibitors, in particular Oseltamivir

have replaced Amantidine as the anti-viral drug of
choice in the treatment of influenza infections.
 Oseltamivir (Tamiflu)
Potent inhibitor of influenza
30 neuraminidase

 Drug of choice for the treatment of influenza

Administered orally
Used for treatment of influenza A and B in persons 18
years or older

Approved for prophylaxis in persons 13 years or older

Its lack of side effects although its costly compared to
amantidine and rimantidine
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It is recommended to be given within 48 hours of the
onset of symptoms
However, some evidence of resistance to oseltamivir
has been reported in H5N1 viruses isolated from
some human H5N1 cases.

 Zanamivir - 
1st neuraminidase inhibitor available for clinical use
Effective against both influenza A and B in persons 12
years or older but not for prophylaxis
Has poor bioavailability hence administered by
inhalation.
Devoid of significant side effects
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in clinical trials
 Its popularity has been eclipsed by oseltamivir
because it cant be taken orally

 Amantidine - this compound inhibit the growth of

influenza viruses in cell culture and in experimental
animals.
Amantidine is only effective against influenza A, and
some naturally occurring strains of influenza A are
resistant to it
The mechanism of action of amantadine is not known.
It is thought to act at the level
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of virus uncoating

The compound has been shown to have both

therapeutic and prophylactic effects

 Rimantadine may be used in place of amantadine for

prophylaxis and the treatment of uncomplicated
influenza A infections.
Rimantidine –
 similar to amantidine but has fewer side effects
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Used for the treatment and prophylaxis of influenza A

infection in persons one year or older

It should be used for uncomplicated influenza A

infections only since it is thought to be less effective
than amantidine
 PREVENTION
Safety precautions with infected
35 birds.
2) Infection control measures with patients.
3) Vaccine
 Immunity to Influenza –
Immunity is induced by the host responses to the virus
haemagglutinin (HA) and to neuraminidase (NA)

Antibody against HA is the most important component

in the protection against influenza viruses.
Serum anti-neuraminidase Ab has also been shown to
contribute protection against influenza infection
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Influenza vaccine must contain both HA and NA

antigens
 Vaccines
1. Inactivated (killed) vaccines
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 Whole virus vaccines:
 The first influenza vaccines to be produced
 Confers protection in 60-90% of vaccinees and the
protection lasts for 1-5 years

 Split virus vaccines:

 less reactogenic but conserving immunogenicity
 Induce fewer side effects in the vaccinees and are just
are immunogenic as whole virus
 Subunit virus vaccines
 contain only the HA and NA
38 antigens
 Induces fewer reactions than those in whole virus
vaccines

2. Live attenuated vaccines - there is experimental

evidence that immunization with live, attenuated
influenza virus vaccines induce a solid immunity than
do inactivated vaccines.
 Avian Influenza

Is an acute
contagious
respiratory disease
affecting both birds
& human.

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 Swine Flu
Also called hog flu & pig flu is an infection by
any one of several types of swine
influenza viruses. 
Swine influenza virus (SIV) is any strain of
the influenza family of viruses that is
endemic in pigs.

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END