ORTHOMYXOVIRUSES

Dr. Tessa Victoria V. Hongco

• Influenza A, B, and C viruses are the most important members of the Orthomyxoviridae
family
• Only influenza A and B viruses cause significant human disease
• Only influenza A can be a zoonosis
• Enveloped, segmented negative-sense ribonucleic acid (RNA) genome
- segmented genome facilitates development of new strains through mutation and
reassortment of the gene segments among different human and animal

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(influenza A) strains
- genetic instability is responsible for the annual epidemics (mutation: drift) and for
influenza A periodic pandemics (reassortment: shift) of influenza infection
worldwide
• Influenza is one of the most prevalent and significant viral infections
- most famous influenza pandemic (worldwide) is the Spanish influenza that swept the
world in 1918-1919, killing 20 to 40 million people
• Influenza viruses cause respiratory symptoms and the classic flulike symptoms of fever,
malaise, headache, and myalgias (body aches)

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Structure and Replication
• Influenza virions are pleomorphic, appearing spherical or tubular and ranging in
diameter from 80 to 120 nm
• envelope contains two glycoproteins, hemagglutinin (HA) and neuraminidase
(NA), and the membrane (M2) protein and is internally lined by the matrix (M1)
protein

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• genome of the influenza A and B viruses consists of eight different helical
nucleocapsid segments, each of which contains a negative-sense RNA associated
with the nucleoprotein (NP) and the transcriptase (RNA polymerase
components: PB1, PB2, PA)
• Influenza C has only seven genomic segments
• genomic segments in the influenza A virus range from 890 to 2340 bases

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• HA forms a spike-shaped trimer; each unit is activated by a protease and is cleaved into
two subunits held together by a disulfide bond
- HA has several functions:
- viral attachment protein, binding to sialic acid on epithelial cell surface
receptors
- it promotes fusion of the envelope to the cell membrane at acidic pH
- it hemagglutinates (binds and aggregates) human, chicken, and guinea pig
red blood cells

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- it elicits the protective neutralizing antibody response
- Changes in HA undergo minor (“drift”) and major (“shift”) changes in receptor
specificity and antigenicity
- Shifts occur only with influenza A virus, and the different HAs are
designated H1, H2…H18

• NA glycoprotein forms a tetramer and has enzyme activity

- NA cleaves the sialic acid on glycoproteins, including the cell receptor
- Cleavage of the sialic acid on virion HA prevents clumping and facilitates the
release of virus from infected cells, making NA a target for two
antiviral drugs, zanamivir (Relenza) and oseltamivir (Tamiflu) 5

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

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• Viral replication begins with the binding of HA to sialic acid on cell surface
glycoproteins
- then internalized into a coated vesicle and transferred to an endosome
- viral envelope then fuses with the endosome membrane allowing uncoating
and delivery of the nucleocapsid into the cytoplasm
- influenza nucleocapsid travels to the nucleus, where it is transcribed
- virus buds selectively from the apical (airway) surface of the cell
- released approximately 8 hours after infection
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Pathogenesis and Immunity
• Disease Mechanisms of Influenza A and B Viruses
-Virus infects the upper and lower respiratory tract
- Systemic symptoms are caused by the interferon and cytokine response to the
virus
- Local symptoms (fever, malaise, headache, and myalgia) result from

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epithelial cell damage, including ciliated and mucus-secreting cells
- Interferon and cell-mediated immune responses (natural killer and T cells) are
important for immune resolution and immunopathogenesis
- Infected people are predisposed to bacterial superinfection because of the loss
of natural barriers and exposure of binding sites on epithelial cells
- Antibody is important for future protection against infection and is specific for
defined epitopes on hemagglutinin (HA) and neuraminidase (NA) proteins
- The HA and NA of influenza A virus can undergo major (reassortment:
shift) and minor (mutation: drift) antigenic changes to ensure the
presence of immunologically naïve susceptible people
- Influenza B virus undergoes only minor antigenic changes
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Epidemiology
• Strains of influenza A virus are classified by the following characteristics:
1. Type (A)
2. Place of original isolation
3. Date of original isolation
4. HA and NA type

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For example, a current strain of influenza virus might be designated
A/Bangkok/1/79 (H3N2), meaning that it is an influenza A virus that was first isolated
in Bangkok in January 1979 and contains HA (H3) and NA (N2) antigens

• Strains of influenza B are designated by

(1) type
(2) geography
(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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• Minor antigenic changes resulting from mutation of the HA and NA genes are called
antigenic drift
- process occurs every 2 to 3 years, causing local outbreaks of influenza A and B
infection
• Major antigenic changes (antigenic shift) result from reassortment of genomes
among different strains, including animal strains
- process occurs only with the influenza A virus

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- Such changes are often associated with the occurrence of pandemics
• In contrast to influenza A, influenza B is predominantly a human virus and does not
undergo antigenic shift

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• Disease/Viral Factors
- large, enveloped virion that is easily inactivated by dryness, acid, and
detergents
- Influenza A infects many vertebrate species, including other mammals and
birds
-Co-infection with animal and human strains of influenza A can generate very
different virus strains by genetic reassortment

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- Transmission of virus often precedes symptoms

• Transmission
- Virus is spread by inhalation of small aerosol droplets expelled during talking,
breathing, and coughing
- Virus likes a cool, less humid atmosphere (e.g., winter heating season)
- Virus is extensively spread by schoolchildren

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• Who Is at Risk?
- Seronegative people Adults: classic flu syndrome
- Children: asymptomatic to severe respiratory tract infections
- High-risk groups: elderly and immunocompromised people, people in nursing
homes or with underlying cardiac or respiratory problems (including
asthma sufferers and smokers)

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• Geography/Season
- Worldwide occurrence
- Epidemics are local; pandemics are worldwide
- Disease is more common in winter

• Modes of Control
- Amantadine, rimantadine, zanamivir, and oseltamivir have been approved for
prophylaxis or early treatment
- Killed and live vaccines contain predicted yearly strains of influenza A and B
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Clinical Syndromes
• incubation period of 1 to 4 days
• “flu syndrome” begins with a brief prodrome of malaise and headache
lasting a few hours
• prodrome is followed by the abrupt and intense onset of fever, chills,
severe myalgias, loss of appetite, weakness and fatigue, sore throat, and

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usually a nonproductive cough
• fever persists for 3 to 8 days
• unless a complication occurs, recovery is complete within 7 to 10 days

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• Diseases Associated with Influenza Virus Infection
Disorder Symptoms
Acute influenza infection in Rapid onset of fever, malaise,
adults myalgia,
sore throat, and nonproductive
cough

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Acute influenza infection in Acute disease similar to that in
children adults but with higher fever,
gastrointestinal tract symptoms
(abdominal pain, vomiting), otitis
media, myositis, and more
frequent croup
Complications of influenza virus Primary viral pneumonia
infection Secondary bacterial pneumonia
Myositis and cardiac involvement
Neurologic syndromes:
Guillain-Barré syndrome
Encephalopathy
Encephalitis
Reye syndrome 14
Laboratory Diagnosis
Test Detects
Cell culture in primary monkey kidney or Presence of virus; limited cytopathologic
Madin-Darby canine kidney cells effects
Hemadsorption to infected cells Presence of hemagglutinin protein on cell
surface

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Hemagglutination Presence of virus in secretions
Hemagglutination inhibition Type and strain of influenza virus or
specificity of antibody
Antibody inhibition of hemadsorption Identification of influenza type and strain

Immunofluorescence, ELISA Influenza virus and antigens in respiratory

secretions or tissue culture
Serology: hemagglutination inhibition, Seroepidemiology
hemadsorption inhibition, ELISA,
immunofluorescence, complement fixation
Genomics: RT-PCR Identification of influenza type and strain
ELISA, Enzyme-linked immunosorbent assay; RT-PCR, reverse transcriptase polymerase chain reaction
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Influenza virus culture in MDCK cell line A. Normal
cell line B. Cell line showing cytopathic effect
(CPE)

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Detection of influenza virus H9N2 in infected
cells. Confirmation of the presence of influenza
BVP01/H9N2 virus at 32 hpi in MDCK-II cells
observed by immunofluorescence assay
(magnification 20×) using influenza A anti-NP
antibodies. A: Infected cells positive for NP
protein; B: Mock control negative for NP staining;
C: Merge between NP and DAPI staining on
infected cells and D: Mock control positive for 17
DAPI staining.
Treatment, Prevention, and Control
• antiviral drug amantadine and its analog rimantadine inhibit an uncoating step of the
influenza A virus but do not affect the influenza B and C viruses.
- target for their action is the M2 protein
• Zanamivir and oseltamivir inhibit both influenza A and B as enzyme inhibitors of
neuraminidase.
- effective for prophylaxis and for treatment during the first 24 to 48 hours after

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the onset of influenza A illness
• airborne spread of influenza is almost impossible to limit
• Natural immunization, which results from prior exposure, is protective for long periods
• Vaccines representing the “strains of the year” and antiviral drug prophylaxis can also
prevent infection
• inactivated subunit influenza vaccines are a mixture of extracts or purified HA and NA
proteins from three or four different strains of virus
• vaccine incorporates antigens of the A and B influenza strains that will be prevalent in
the community during the upcoming winter
• Annual vaccination is the most important measure to prevent influenza and its
complications 18

- It is recommended for all people aged 6 months and over

• Vaccination is routinely recommended for all individuals and especially persons older
than 50 years, health care workers, pregnant women who will be in their second or
third trimester during flu season, people living in a nursing home, people with chronic
pulmonary heart disease, and others at high risk

• The WHO recommends that quadrivalent vaccines for use in the 2022-2023
influenza season in the northern hemisphere contain the following:
• Egg-based vaccines

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• an A/Victoria/2570/2019 (H1N1)pdm09-like virus;
• an A/Darwin/9/2021 (H3N2)-like virus;
• a B/Austria/1359417/2021 (B/Victoria lineage)-like virus; and
• a B/Phuket/3073/2013 (B/Yamagata lineage)-like virus

• Cell culture- or recombinant-based vaccines

• an A/Wisconsin/588/2019 (H1N1)pdm09-like virus;
• an A/Darwin/6/2021 (H3N2)-like virus;
• a B/Austria/1359417/2021 (B/Victoria lineage)-like virus; and
• a B/Phuket/3073/2013 (B/Yamagata lineage)-like virus.
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• The WHO recommends that trivalent vaccines for use in the 2022-2023 influenza
season in the northern hemisphere contain the following:
• Egg-based vaccines
• an A/Victoria/2570/2019 (H1N1)pdm09-like virus;
• an A/Darwin/9/2021 (H3N2)-like virus; and
• a B/Austria/1359417/2021 (B/Victoria lineage)-like virus.

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• Cell culture- or recombinant-based vaccines
• an A/Wisconsin/588/2019 (H1N1)pdm09-like virus;
• an A/Darwin/6/2021 (H3N2)-like virus; and
• a B/Austria/1359417/2021 (B/Victoria lineage)-like virus

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• A live attenuated influenza vaccine (LAIV) is also available for administration as a nasal
spray instead of a “flu shot”
- All nasal spray influenza (flu) vaccines are quadrivalent, meaning they are
designed to protect against four flu viruses: an influenza A(H1N1) virus, an influenza
A(H3N2) virus and two influenza B viruses
- vaccine is restricted to infecting the nasopharynx and will elicit a more natural
protection, including cell-mediated, serum antibody, and mucosal-secretory
immunoglobulin (Ig)A antibody

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- vaccine is only recommended for people aged 2 to 50 years

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