S3November 17, 2010

Lec 11: Orthomyxovirus and Paramyxovirus by Dr. Madrid

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• Each segment
encodes a different viral protein

A FRIENDLY NOTE FROM THE NEW AND IMPROVED

MICROBIOMAN:
Viral Proteins and Function
To facilitate you in studying this trans, enclose in textbox RNA Protein Function
(like this) were lifted from Jawetz. Hope this helps. 
1—3 Pol B2, B1, A RNA synthesis, core
Legend:
RT- Respiratory tract
HAI- hemagglutin inhibition 4 Hemagglutinin Attachment
IF- Immunofluorescence
5 Nucleoprotein RNA synthesis, Core receptor
Happy Aral! 
ORTHOMYXOVIRUS 6 Neuraminidase Release of virus
• Highly infectious viral illness 7 M-1, M-2 Scaffolding, ion channel
• Epidemics reported since at least 1510
• at least 4 pandemics in the 19th century 8 NS-1, NS-2 Regulates mRNA splicing
• First isolated in 1933

4 Genera: Influenza A, B, C and Thogotoviruses (found in Influenza Virus – Surface Spike: GLYCOPROTEINS
mosquitoes, ticks and banded mongoose) HA & NA

IMPORTANT PROPERTIES OF ORTHOMYXOVIRUSES • High frequency of variations, caused by

HA & NA
Virion: Spherical, pleomorphic, 80–120 nm in diameter • Variations result in the spread of new
(helical nucleocapsid, 9 nm) serologic types
• Hemagglutinin
Composition: RNA (1%), protein (73%), lipid (20%),
○ Binds to receptor (a trisaccharide
carbohydrate (6%)
of sialic acid, galactose & glucose
Genome: Single-stranded RNA, segmented (eight on host cells)
molecules), negative-sense, 13.6 kb overall size
○ Sialic acid fits into pocket on top
Proteins: Nine structural proteins, one nonstructural of HA; pocket highly conserved
Envelope: Contains viral hemagglutinin (HA) and but the rest of HA is not
neuraminidase (NA) proteins
Replication: Nuclear transcription; capped 5' termini of
cellular RNA scavenged as primers; particles mature by
budding from plasma membrane
Outstanding characteristics:

Orthomyxovirus

• Influenza Viruses Type A, B and C

○ Subtypes due to antigenic variations of
surface glycoprotein’s hemagglutinin
(HA) and neuraminidase (NA)
Paramyxoviruses

• Parainfluenza and Respiratory Syncytial Virus

(RSV)

Influenza Virus Structure

• Genome:
○ Influenza A & B- 8 individual
segments;
○ Influenza C – 7 segments

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 ○ HA is the target of neutralizing • Influenza A & B have multiple
antibodies strains based on variation of HA
and NA
• Neuraminidase • Type, host origin, geographic
○ A “square-topped, mushroom-like origin, strain # & year isolated
projection” • Antigenic descriptions (HA and
○ Important for release of virion NA)
from cells • 3 subtypes of HA (H1-H3) and two
subtypes of NA (N1 and N2).
STRUCTURE Examples:
○ A/Hong Kong/03/68
VIRUS REPLICATION (H3N2)
○ A/swine/Iowa/15/30
(H1N1)

Antigenic Variation

• Antigenic DRIFT- selection of mutants

less susceptible to most common
antibodies
• Antigenic SHIFT -new antigenic type
unrelated to earlier types
○ Occurs infrequently and only with
Influenza A
○ Influenza A - major shifts:
 H1N1 (1918)
 H2N2 (1957)
 H3N2 (1968)
 H1N1 (1976)

UNIQUE PROPERTIES OF ORTHOMYXOVIRUSES

• division into 8 separate segments

Virus binds & enters cell (1)  Endosome fuses with facilitates development of new strains by
lysosome Acid stimulates HA fusion activity mutation and reassortment of the gene
Replication begins ~1 hour later  Nucleus is important segments between different human and
in replication: does NOT occur in enucleated cells  5’ animal strains of virus
ends of nuclear transcripts used as viral mRNA  ○ responsible for the animal
Polymerase A & NP bind to RNA; stop polyadenylation & influenza epidemics
capping  Uncapped RNA makes genome RNA  • influenza epidemics (mutation : drift) and
Proteins associate with cell membrane  Virus is periodic pandemics (reassortment : shift)
released as it forms – no fully formed virions inside cells of influenza infection worldwide
• Enveloped virion with a genome of 8
different (-) RNA nucleocapsid segments
• The hemagglutinin (H) glycoprotein is the
VAP, fusion protein, and elicits
neutralizing, protective antibody
responses
• mutation-derived changes in H causes
antigenic changes of minor(drift) or
major(shift) degree
• Shift
○ MAJOR change
○ Results to new subtype
○ Caused by exchange of gene
segment
Host Range ○ pandemic
 e.g. Antigenic Shift
• Influenza A: humans and other species ✔ H2N2 1957-67
(swine, horses, birds) ✔ H3N2 1968
• Influenza B and C only: humans • Drift
• Influenza C has only one strain ○ MINOR change
○ same subtype
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 ○ caused by point mutation in gene ○ avian strain H5N1 , transmit
○ epidemic directly from chickens to humans
 e.g. Antigenic drift
✔ 1997- INFLUENZA VIRUS EPIDEMIOLOGY
A/Wuhan/359/95
(H3N2) • Influenza outbreaks occur every year,
✔ 1997 to 1998- extent and severity vary with antigenic
A/Sydney/5/97(H3 composition
N2) • Influenza A outbreaks most common,
• NEURAMINIDASE (N) Influenza B outbreaks less extensive and
○ hydrolyze the protective mucous less severe
coating on the respiratory tract • Influenza C infrequently associated with
○ assist in viral budding and release human disease – Antibodies against
influenza C present in serum,
○ prevent viruses from sticking
asymptomatic infection
together
• Epidemics – when non-immune/partially
○ participate in host cell fusion
immune are infected
• Influenza transcribes and replicates its • Epidemics occur most often during winter
genome in the target cell nucleus but • # of US citizens killed during 1918
assembles and buds from the plasma pandemic – 548, 452
membrane • # of US citizens killed during WWI –
• The antiviral drugs inhibit an uncoating 53,513 (10x fewer)
step and most likely target the M2 protein • India 12.5 million, world wide 20 million
• The segmented genome promotes • Infects many vertebrates species
genetic diversity caused by mutation and including mammals and birds
reassortment of segments upon infection • Co-infection with animal and human
with 2 different strains strains of virus – generate different
strains by genetic reassortment
ORTHOMYXOVIRUS PANDEMICS
Transmission
Year Subtype • Inhalation of small aerosol droplets
(coughing and sneezing)
1890 H2N2 • Confined populations – nursing homes,
classrooms, ships
1900 H3N8
Physical Characteristics
1918 H1N1(Spanish
) • Withstands slow drying at room temp
based on articles
1957 H2N2 (Asian) • Demonstrated in dust after an interval of
1968 H3N2 ( Hong 2 weeks
• Survive for several weeks at 4oC when
Kong)
contaminated in infected tissue immersed
1977 H3N2 + H1N1 in glycerol saline
• survive in seawater
1990 H3N2 + H1N1 • preserved for long periods at –70oC
• inactivate after exposure to heat for 30
NOMENCLATURE min at 56°C : 90 min
• inactivated by 20% ether in the cold,
• host of origin/geographical origin/ strain phenol, formaldehyde, salts of heavy
number/ year of isolation/ antigenic metals, detergents, soaps, halogens
description of hemagglutinin and
neuraminidase
• e.g. A/swine/Iowa/3/70(H1N1) Disease Mechanism
A/Scotland/42/89 (H3N2)
• Virus can establish infection of URT/LRT
ORTHOPARAMYXOVIRUS • Systemic symptoms are due to the
interferon and lymphokine response to
• H 15 ; N 9 the virus. Local symptoms are due to
• H1-H3 ; N1-N2 epithelial cell damage, including ciliated
○ found in epidemic/pandemic and mucus secreting cells
viruses from humans • Interferon and Cell mediated immune
• 1997 outbreak in Hong Kong (CMI) response (NK and T cell) are

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 important for both immune resolution and
immunopathogenesis
• Infected individuals are predisposed to
bacterial superinfection because of the
loss of these natural barriers and
induction of bacteiral adhesion to
epithelial cells
• Antibody is important for future
protection against infection and is
specific for defined epitopes on H and N
• The H and N of influenza A can undergo
major (reassortment,shift) and minor
(mutation,drift) antigenic changes to
ensure the (+) of immunologically naïve,
susceptible individuals
• Influenza B undergoes only minor
antigenic changes
• Exclusively in 2-6 y/o, mostly with
influenza B, less common with INF A, VZV,
measles, rubella & poliovirus
• Symptoms: encephalitis, mental status
changes (ranging from lethargy to coma,
including delirium and seizures),
hepatomegaly, increased levels of blood
ammonia and bilirubin (moderate, so no
jaundice), hypoglycemia
• Fatty liver changes associated with
aspirin treatment and Viral infection
• Decreased incidence since warnings of
aspirin use in children with acute viral
respiratory infections; mortality rate
initially ~40%, now<10%
Immunity

Clinical Infections • Depends on immunity to previous variant

circulating in population and on
• Symptoms appear suddenly 1-2 days relatedness of the two variants
after exposure • Most epidemics due to antigenic shifts
• Major symptoms: rapid rise in body temp, that produce subtypes distantly related to
to 102°F, myalgias and on occasion sore previous types
throat, headache, cough and nasal • IgA, serum IgG and cellular immunity
congestion important
• Other symptoms: mark lassitude, • Particular subtype of infecting virus is of
moderate anorexia long duration
• Epithelial cells of upper and lower • Related to the amount of local Ab (IgA) in
respiratory tract the mucous secretion of the RT + specific
• Influenza A incubation period: 1-4 days (2 IgG serum Ab concentration
days average) • Immunity to infection (type A) subtype
○ Adults : sudden onset of fever, specific
malaise, headache, myalgia,
anorexia, sore throat, dry cough Diagnosis
○ Children : higher fever, GI
symptoms (abdominal pain, Test What it
vomiting), OM , myositis, croup detects?
• Influenza B: >milder 3 day febrile illness
with predominantly systemic symptoms; Cell culture in PMK or Virus
more GIT involvement “gastric flu” MDCK
• Influenza C: afebrile URTI; confined to
young children Hemadsorption to Hemadsorbing
infected cells virus
Complications
Ab inhibition of Virus, type strain
• Pneumonia – primary influenza viral hemadsorption
pneumonia (rare, depending on strain)
• Secondary bacterial pneumonia or mixed IF, ELISA Virus antigens
viral and bacterial pneumonia
(pneumococci – S. pneumoniae, HI Virus, type and
staphylococci – S. aureus, H. influenza) strain
• Myositis and cardiac involvement
• Neurological syndromes Serology: HI, HAI, ELISA, seroepidemiology
○ GBS
IF, CF
○ encephalopathy
○ encephalitis
○ Reye’s syndrome Treatment
Reye’s Syndrome • Amantadine
○ ineffective against Influenza B
• Sequelae of Influenza
and Influenza C; it is relatively
nontoxic, but might stimulate

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 CNS; dizziness & insomia, ✔ Elderly (> 50 yrs)
particularly in elderly. ✔ > 6 mos with chronic illness
• Rimantadine ✔ Residents of long term care facilities / health care
○ block ion channels in the providers
envelope ✔ Employees of long term facilities
 preventing the pH ✔ Household members of high-risk persons
changes that precede the ✔ Providers of essential community services
membrane fusion step ✔ Pregnant women
essential for nucleocapsid ✔ Persons 6 mos to 18 yrs – receiving ASA
release (Influenza A) Virion: Spherical, pleomorphic, 150 nm or more in diameter
treatment
✔ (helical illnesses:
Chronic nucleocapsid, 13–18 nm)
• Amantidine & Rimantidine are weak Diameter:
○ pulmonaryRange: disease
100 – 800 nm; Average: 125 – 250
organic bases; block uncoating of INF A & nm○ Heart disease
prevent viral replication ○ Metabolic
Composition: RNA (1%), protein (73%), lipid (20%),
• M2 Matrix protein binding; resistance is carbohydrate
○ Renal (6%) dysfunction
due to mutations in genes encoding M2 ○ Hemoglobinopathies
Genome: Single-stranded RNA, linear, nonsegmented,
• Relieves symptoms only (doesn’t alter negative-sense, noninfectious, (HIV)
○ Immunosuppression about 15 kb
disease course) ✔ Foreign travelers
Proteins: Six to eight structural proteins
✔ Students
Envelope: Contains viral glycoprotein (G, H, or HN) (which
Neuraminidase inhibitors: (A and B) ✔ Anyone who wishes to reduceorthe likelihood activity)
of
• Zanamivir sometimes carries hemagglutinin neuraminidase
becoming
and fusionill(F)from influenzavery fragile
glycoprotein;
○ poor bioavailability
○ inhalation 2x a day for 5 days Replication: Cytoplasm; particles bud from plasma
membrane
PARAMYXOVIRUSES
○ Relenza inhaled, improvement if
taken within 2 days of flu Outstanding
IMPORTANT characteristics:
PROPERTIES OF PARAMYXOVIRUSES
symptoms
• Oseltamivir
○ oral. 2x a day for 5-7 days
○ Tamiflu (not for infants) inhibits
NA
○ OTC analgesics helps reduce
headache, fever/myalgia
• Active against Both Influenza A and
Influenza B

Prevention

• Vaccines: inactivated virus produced in

eggs; ~70% effective (lack of local IgA,
cell-mediated responses)
• Complications: hypersensitivity reactions
in persons with egg allergies; Guillain-
Barré Syndrome
• Strain of virus important; antigenic
variation makes vaccines obsolete, WHO • 3 Genera
surveillance teams look for new variants ○ Paramyxovirus
(best guess)  Parainfluenza 1-4
• Immunity is of short duration: 1-3 years • Mumps
• Mild tenderness/redness in 30% of • Newcastle disease Virus
vaccines, fever & mild systemic (NDV)
symptoms in up to 5% of vaccinees • Simian virus 5 (SV5)
• Additive protective effect of vaccine & ○ Morbillivirus
amantidine  Measles
 Canine distemper virus
CONTROL  Rinderpest –equine; several
strains of seals, dolphins and
Immunization – 3 virus strains porpoises
○ Pneumovirus
• 2 type A + 1 type B  Respiratory Syncytial virus (RSV)
• Provoke a good local (IgA) antibody
response

Recommended for:

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 • Transmitted in respiratory droplets and
initiate infection in the respiratory tract
• CMI causes many of the symptoms but is
essential for control of infection
• Easily deformed by external forces
• May assume a variety of shapes
HISTORY • Break up more easily

• 1990s: Hendra and Nipah REPLICATION

○ discovered in Australia and SEA
○ animal viruses occasionally Entry into cell:
transmitted in man • Attachment to host cells via the
hemagglutinin (HN) protein
• F protein involved in fusion
Replication:
• Occurs in cytoplasm of cell
• Viruses have RNA-dependent RNA
polymerase
• Release is by budding from the cell
surface
Fate of host cell:
• Syncitium formation common
• Cytoplasmic eosiniphilic inclusions: site of
viral synthesis; contain nucleocapsids and
viral proteins (RSV & Measles)
Proteins • Usually minimal effects on host cell
metabolism, unless extensive cell fusion
• Enveloped may contain 2 glycoproteins: occurs
• HN – hemagglutinin and neuraminidase
(mumps) activity
• F – hemolytic and cell fusion activity MEASLES VIRUS
• M – matrix, inner layer of envelope
• NP – nucleoprotein Epidemiology
• L – large protein
• P – polymerase • Transmission : inhalation via large droplet
aerosol
Unique Features • Contagion period precedes symptoms
• Host range is limited to humans
• Large virion consisting of a (-) RNA • Only 1 serotype
genome in a helical nucleocapsid • Immunity is lifelong
surrounded by an envelope containing a
Disease Mechanism
VAP
• Infects epithelial cells of RT
○ HN- paramyxovirus; H- • Spreads systemically in lymphocytes and
morbillivirus G: pneumovirus) and by viremia
a fusion glycoprotein (F) leading • Replicates in cells of the conjunctiva,
to syncytia formation Respiratory tract, GIT, Urinary tract,
lymphatic system, blood vessels and CNS
• The 3 genera can be distinguished by the • Rash is caused by T-cell response to virus
activities of the VAP : infected epithelial lining in the capillaries
○ HN of paramyxovirus has • CMI is essential to control of infection,
antibody not sufficient due to measles
hemagglutinin and
ability to spread cell to cell
neuraminidase activity
○ H of morbilluvirus has
• Sequelae in the CNS may result from
immunopathogenesis (postinfectious
hemagglutinin activity measles encephalitis) or development of
○ G of pneumovirus lacks these defective mutants (subacute sclerosing
activities panencephalitis, SSPE)
• Virus replicates in the cytoplasm
• Virus penetrates the cell by fusion with Clinical consequences
the plasma membrane
• Viruses induce cell-cell fusion, causing Disorder Symptoms
multinucleated giant cells

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Measles Koplik's spots, rash extends to the body
and extremities Disease Mechanisms

Atypical Rash prominent in distal areas • 4 serotypes

measles • Infection is limited to RT with URTI being
the most common, but significant disease
SSPE CNS manifestations: personality, can occur upon LRTI
behavior, memory changes, myolonic • NOT systemic and do not cause viremia
• Diseases:
jerks, spasticity, blindness
○ Cold like symptoms, bronchitis,
bronchiolitis, croup
Complications ○ Parainfluenza 1, 2, or 3 in:
 Infants – more severe,
• Laryngitis appearing as
• Otitis media bronchiolitis, pneumonia,
• Encephalitis – 7 to 10 days after onset croup
• Pneumonia (Laryngotracheobronchiti
○ giant cell pneumonia without rash s - LTB)
○ (-) CMI  Older children and adult:
milder infections
Clinical Diagnosis • Infection induces protective immunity of
short duration
• Clinical specimens: RT secretions, urine, blood,
Clinical Syndromes
brain tissues
• Stain: Multinucleated giant cells with
• Variety of syndromes, primarily in infants
cytoplasmic and inclusion bodies
and young children
• SSPE:
○ high levels of measles antibody in the
• LTB (croup) – sore throat, hoarseness,
watery discharge and cough.
blood and CSF
○ Severe cases – fever persists,
○ eosinophilic inclusion bodies in the brain
with worsening discharge and
sore throat
Treatment: Supportive; Vitamin A
• Bronchiolitis and pneumonia – associated
with serotype 3, infects infants less than
Prevention
1 year old.
• Live attenuated measles vaccine
• (+) exposure  Ig within 6 days of
• Common cold in subclinical form – usual
result of adult infection with any type
exposure
Diagnosis
PARAINFLUENZA VIRUS
• Isolation from nasal washings and
Epidemiology
respiratory secretions
• Transmission – respiratory secretions • IF, HI
○ person to person
Treatment : mostly supportive
○ respiratory droplet
• Primary infection : < 5 yrs
• Upper respiratory tract illness –
• (+) reinfection
symptomatic therapy only
• Closed populations including young
• Sinusitis, otitis, bacterial bronchitis –
children are at risk
appropriate antibiotics for post-viral
• Type 3 is most prevalent serotype
bacterial infection
• Parainfluenza – cause human respiratory
disease throughout the year, but
• No specific antiviral therapy – ribavirin
has activity against parainfluenza viruses
especially during Fall and Winter
in vitro and is being tested
• No effective vaccines available
Antigenicity
MUMPS
• Four major serotypes, based on HN, F and
NP protein antigens
• Acute inflammation of parotid glands
• Types 1, 2 and 3 related, type 4 not
related
Epidemiology
• Most adults have circulating Ab’s
necessary to prevent primary infection
• Transmission
• Reinfection by same type is common
○ person to person
despite presence of Ab against it
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 ○ respiratory droplets • Transmission : inhalation of droplet
• Present in respiratory secretions x 7 aerosols
days before illness • Peaks in December
• One serotype • Yearly worldwide epidemics in infants and
• Infects only humans young children
○ Major impact is during first 6
Antigenicity months of life, ~30% develop
Ab’s in first year, 95% by 5 years
• Much like parainfluenza virus, surface HN ○ Initial site of virus multiplication is
glycoprotein, a surface F gp, and an epithelium of upper respiratory
internal NP tract then spreads to lower tract
• Single antigenic type (MMR) (bronchi, bronchioli, lung
parenchyma)

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ppease his justice, to demonstrate his sovereignity. But one of the sweetest reasons God saved you is
, your photo would be in it. He sends you flowers every spring and sunrise every morning. Whenever

• Mumps specific IgM antibody HI, ELISA, IF
Treatment: Supportive
Prevention: Live attenuated vaccine – Jeryl Lynn strain
RESPIRATORY SYNCYTIAL VIRUS (RSV)
• Major cause of lower respiratory tract
disease in infants and young children,
especially in closed situations. Adults
show mild or no symptoms; does not
spread in older children and adults
Epidemiology
• Contagion period preceded symptoms
and may occur in the absence of
symptoms
• Cell fusion and aspiration may be
involved in spreading
Antigenicity
• Three minor types with high degree of
cross-reactivity
• Immunity is short lived and dependent
upon secretory IgA in nasal secretions,
not on circulating IgG concentrations
Disease Mechanism
• Localized infection of RT
• Does not cause viremia or systemic
spread

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 • Pneumonia results from cytopathic effect
of virus (including syncytia)
• Bronchiolitis most likely mediated by
host’s immune response
• Narrow airways of young infants readily
obstructed by virus-induced pathology
• maternal antibody does not protect infant
from infection
• natural infection does not prevent
reinfection
• vaccination increases severity of
subsequent disease

Clinical Infections

• Bronchiolitis – difficulty of breathing with

evidence of obstructed airway (can be
distinguished from asthma), noisy
breathing
• Pneumonia – pulmonary infiltrates are
more prominent, can involve swelling of
alveolar lining cells and interstitial
inflammation
• Respiratory failure can occur
• Bronchiolitis in childhood may be linked
to chronic lung disease later in life
Diagnosis

• Human and simian cell cultures

• Syncytia formation in culture
• Difficult to isolate in cell culture IF, EIA

Prevention: No vaccine available

Treatment: Ribavirin via inhalation
Control: Standard precaution

------------------------------------------end of
trans-------------------------------------------------

Ang nag-uumapaw sa impormasyon na trans na ito ay

inihahandog ng:

MICROBIOMAN

(From L to R): Paulfie, Turay, Edo, Nina, Teacher

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