Togaviridae

Section of Microbiology and

Parasitology
 Togaviruses

(+)sense RNA Viruses

Family Genus Type Species Hosts

Togaviridae Alphavirus Sindbis virus Vertebrates

Rubivirus Rubella virus Vertebrates

 Togaviridae
1. Alphavirus
a) Sindbis
b) Semliki Forest
c) Venezuelan equine
encephalitis
d) Eastern equine
encephalitis
e) Western equine
encephalitis
f) Chikungunya

2. Rubivirus
Togavirus:
morphology
 Toga: from Latin toga
“cloak”  Envelope-
 Capsid: 40-70 nm – Tightly bound to
diameter capsid
– Enveloped; Spherical – 2 peplomers: E1
– Icosahedral and E2 are
responsible for viral
nucleocapsid
specificity
– Comprised of
– Envelope acquired
multimers of 1 gene by budding
product
 Togaviruses

 Genome : positive sense, single stranded RNA

that is 10,000-12,000 nucleotides long
• The 5'-terminus carries a methylated nucleotide
cap and the 3'-terminus has a polyadenylated tail
• ~11.7kb, 4-8% total weight of particle
• Nonstructural proteins encoded at the 5' end
• Togaviridae make use of a subgenome strategy to
synthesize viral proteins
 The (+) sense genomic RNA ('49S'
= 11.7kb) acts directly as mRNA
 partially translated (5' end) to
produce nonstructural proteins 
these proteins are responsible for
replication, forming a
complementary (-) strand 
template for further (+)strand
synthesis
 2 species of (+) RNA are
synthesized:
a. full length genomic RNA
b. sub-genomic mRNA ('26S' = 4.1kb)
– Translation of the newly
synthesized sub-genomic RNA
results in production of structural
proteins
 Assembly occurs at the cell
surface
 envelope is acquired as the virus
buds from the cell
Replication
Alphaviruses
 Togaviridae: Alphavirus genus
Important members Virus properties
Chikungunya virus, • Spherical, 70 nm in
Eastern equine encephalitis virus, diameter
Mayaro virus, • Nucleocapsid has 42
O’Nyongnyong virus, capsomeres
Ross River virus, • Genome: (+) sense ss
Semilki Forest virus, RNA, 9.7-11.8kb in size
Sindbis virus, • Enveloped
Venezuelan equine encephalitis • Replication: cytoplasm
virus. • Assembly: budding
Western equine encephalitis virus through host cell
membrane
 Togaviridae:
Togaviridae: Alphavirus
Alphavirus
 Consists of about 30  Envelope: contains 2
viruses; 70 nm diameter glycoproteins
 Genome: single-stranded, +  establish persistent infections
sense RNA (single 42S in mosquitoes and are
transmitted between
strand of approximately 4 x vertebrates by mosquitoes
106 daltons that is capped and other blood-feeding
and polyadenylated) arthropods
 icosahedral nucleocapsid
 Distribution: worldwide
enclosed in a lipid-protein
 Inactivated by acid pH, heat,
lipid solvents, detergents,
envelope bleach, phenol, 70% alcohol
– The single capsid protein (C and formaldehyde
protein) has a molecular
weight of approximately
30,000 daltons
Alphavirus structure
 Envelope: lipid bilayer derived
from the host cell plasma
membrane
– contains 2 viral glycoproteins (E1
and E2) of molecular weights of
48,000 to 52,000 daltons.

 The only proteins in the

envelopes of alphaviruses are the
viral glycoproteins, each
anchored in the lipid at or near
their C-terminus.

 On the virion surface, E1 and E2

are closely paired, and together
form trimers that appear as
"spikes" in an orderly array.
 Alphaviruses: replication
 Genomic length and subgenomic mRNAs are
produced during transcription
 The genomic length transcript produces a
precursor polyprotein encoding the nonstructural
proteins needed for viral replication.
 The subgenomic mRNAs encodes structural
proteins
 The proteins are elaborated by posttranslational
cleavage
 Alphaviruses replicate in the cytoplasm and
mature by budding nucleocapsids through the
plasma membrane
 Genomic organization of alphaviruses

 nonstructural proteins (nsP) are translated from the

genomic RNA as a polyprotein that is processed into 4
proteins by a viral protease present in nsp2.
 structural proteins are translated from a subgenomic
26SmRNA as a polyprotein that is processed by a
combination of viral and cellular proteases into a
capsid protein (C), 3 envelope glycoproteins (E3, E2,
and E1), and a membrane-associated protein named 6K

Genome RNA
Cap An
nsP1 nsP2 nsP3 nsP4 Subgenomic 26S mRNA
cap An

C E3 E2 6K E1
 Alphavirus: functions of nonstructural Proteins

 nsP1: synthesis of minus strand RNA

 nsP2: regulation of minus strand synthesis;
 initiation of subgenomic RNA synthesis;
 processing of the NS polyprotein  
 nsP3: unknown 
 nsP4: viral polymerase
 Structural Proteins: alphaviruses
capsid E3 E2 6k E1
Functi Forms Contains signal Surface May act Surface
on nucleoc sequence for glycoprote as signal glycopr
apsid protein in; sequence; otein
translocation; interacts important
ultimate with host in virus
function receptors, assembly
unknown (not functions and
present in all in budding.
mature attachment
alphaviruses)
Multiplication
 Genomic RNA serves as
mRNA for nonstructural
proteins (e.g., RNA-
dependent RNA
polymerase) which are
encoded in the 5' two-
thirds of the genome.
 Complementary RNA,
made from genomic RNA,
serves as a template for
progeny genomic RNA.
 A subgenomic mRNA
representing the 3' one-
third of the genome
encodes the structural
proteins.
 Lifecycle: Alphavirus
1. Virus attaches to host receptors though E2 glycoprotein
endocytosed into vesicles in the host cell.
2. Fusion of virus membrane with the vesicle membrane 
nucleocapsid released into the cytoplasm 
uncoating RNA genome is released
3. Positive-sense genomic ssRNA used as the messenger
RNA for the translation of the nonstructural proteins (p270
and p230 polyproteins.)
4. A negative-sense complementary ssRNA is synthesized
using the genomic RNA as a template.
5. New genomic RNA is synthesized using the negative-sense
RNA as a template.
A subgenomic RNA (26S RNA) containing the last third of the genome
is also synthesized.
6. Structural proteins (p130 polyprotein) are translated from
the 26S subgenomic RNA.
7. Capsid protein cleaved autocatalytically from the p130
polyprotein; remainder of the polyprotein moves into the ER
membrane
Post-translational modifications (glycosylation and
acylation) and further cleavage occurs, yielding four
separate proteins (E3, E2, 6K, and E1).
8. Membrane-embedded proteins move to the surface of the
cell  virus assembly occurs
C-terminal region of the E2 protein interacts with the
nucleocapsid; the N-terminal region of E2 interacts with E1
to give heterodimer "spikes“  these spikes associate in
trimers to give larger spike formations
assembled virions are released from the cell
 Envelope proteins formed by posttranslational cleavage
are glycosylated and translocated to the plasma
membrane.
 Virion formation occurs by budding of preformed
icosahedral nucleocapsids through regions of the plasma
membrane containing E1 and E2 glycoproteins
 virus Clinical syndrome vector host distribution
EEE encephalitis mosquito birds Americas

WEE encephalitis mosquito birds North America

VEE Febrile illness, mosquito Rodents, Americas

encephalitis horses
Chikungu Febrile illness, rash, mosquito Primates, Africa, India,
nya arthralgia humans Southeast asia
Mayaro Febrile illness, rash, mosquito Primates, South America,
arthralgia humans Trinidad
Onyongny Febrile illness, rash, mosquito primates Africa
ong arthralgia
Ross river Febrile illness, rash, mosquito Mammals, Australia, Pacific
arthralgia humans
SIndbis Febrile illness, rash, mosquito birds N. europe, Africa,
arthralgia Asia, Australia
Semliki Febrile illness, rare mosquito birds Africa
forest encephalitis
Alphavirus: main host are domesticated and wild animals.
 virus is taken up in the animal host's blood by mosquitoes
– virus replicates within the mosquito vector before it is introduced
into a human host through mosquito bite.

 virus is introduced into the capilarriesreplicates in the

vasular endothelium  spread to other target tissues
including muscles, joints, skin, and brain
 Pathogenesis
 transmitted via infected mosquitoes
– Virus is transmitted from the salivary glands of the
mosquito to the bloodstream of the vertebrate host.
– Virus travels to the skin and reticuloendothelial system
(spleen and lymph nodes), where the primary infection
occurs  then viraemia follows  systemic infection
– Can involve CNS (esp. encephalitis), skin/bone
marrow/blood vessels (haemorrhagic fevers)

 Infection with seroconversion in the absence of clinical

disease is common, but disease can be incapacitating and,
in cases of encephalitis, occasionally fatal

 Virus is eliminated by the immune system but arthritis or

central nervous system impairment may persist for weeks.
 Alphaviruses: Host Defenses
 Initial resistance is conferred by
nonspecific defenses such as interferon.
 Antibodies are important in recovery and
resistance
– T-cell responses are also involved.
 Lasting protection is generally restricted
to the same alphavirus and is associated
with, but not solely attributable to, the
presence of neutralizing antibodies.
 Alphaviruses: Epidemiology
 maintained in nature by mosquito-vertebrate-
mosquito cycles

 Restricted interactions between viruses, vector

species, and vertebrate hosts tend to confine the
geographic spread of alphaviruses

 Occasionally, a virus may escape its usual ecological niche

and cause widespread epizootics (Venezuelan equine
encephalitis virus) or urban epidemics (chikungunya virus)
– Human infections are seasonal and are acquired in endemic areas.
 Epidemiology
 EEE virus: fresh water
swamps in the eastern US  WEE virus (widespread in
– causes sporadic equine & rare the US and Canada)
human cases – outbreaks of equine and
 Small human outbreaks may human disease in western
occur and southwestern states.
– Since Culiseta melanura – principal vector: Culex
mosquitoes usually do not feed tarsalis ,a common
on humans, transmission to mosquito, especially in
horses and humans is irrigated regions
potentiated when less fastidious
Aedes species feed upon an
adequate natural reservoir of
infected birds.
 VEE virus: Eight or more antigenic subtypes
– Can be transmitted by at least 10 different species of mosquitoes,
including Culex and Aedes species
– Birds do not seem to play an important reservoir role in nature.
– Enzootic subtypes of relatively low equine virulence exist in South and
Central America and Florida.
 Epidemiology
 Chikungunya virus
– exists in Africa in a forest cycle involving baboons and
other primates and forest species of mosquitoes
– can also be transmitted in a human-mosquito-human
cycle by Aedes aegypti
 massive epidemics in Africa, India, and Southeast Asia
– is endemic throughout much of south and Southeast
Asia
 Mayaro virus
– This antigenically similar virus exists in the Amazon Basin.
– Its cycle involves new world primates and hematophagous
mosquitoes; causes outbreaks of human disease through
exposure to the forest cycle.
 Ross River virus
– is endemic in Australia and has spread in epidemic form to several
islands of the Western Pacific.
 Epidemiology
 O’nyong nyong virus
– Found in Africa
– Vectors:mosquitos - Anopheles spp. , A. funestus, A.
gambiae

 Semliki forest virus:

– isolated from mosquitoes in the Semliki Forest,
Uganda; pathogenic for laboratory animals and
man

 Sindbis virus
– Common in Africa, Asia, Australia, Middle East,
Eastern Europe, Scandinavia and the Commonwealth
of Independent States
– transmitted by a mosquito vector
Alphavirus: clinical manifestations

 alphaviruses can cause various syndromes,

ranging from benign febrile illnesses to
severe systemic diseases with hemorrhagic
manifestations or major organ involvement.
 The neurotropic alphaviruses can produce
severe destructive central nervous system
disease with serious sequelae.
 Several alphaviruses (chikungunya, Mayaro,
and Ross River) cause painful arthritis that
persists for weeks or months after the initial
febrile illness.
Alphavirus: clinical manifestations
Human illness disease patterns.
 Chikungunya virus is the prototype for those
causing an acute (3- to 7-day) febrile illness with
malaise, rash, severe arthralgias, and
sometimes arthritis.
 O'nyong'nyong, Mayaro, and Ross River
viruses, which are closely related (antigenically)
to chikungunya virus, cause similar or identical
clinical manifestations
 Sindbis viruses cause similar but milder
diseases known as Ockelbo (in Sweden),
Pogosta (Finland), or Karelian fever (Russia).
 Alphavirus: clinical manifestations
 viremia coincident with abrupt onset of fever,
chills, malaise, and joint aches.
– viremia subsides in 3 to 5 days, and antiviral antibodies
appear in the blood within 1 to 4 days of the onset of
symptoms

 A macular-papular rash typically develops around

the third to fifth day of illness, when the patient is
defervescing.

 migratory arthralgia (characteristic feature),

involves mainly the small joints
– occurs more prominently in adults than children.
– severe cases: joints are swollen and tender
Alphavirus: Diagnosis

 Diagnosis is suggested by clinical evidence

and by known risk of exposure to virus.
 Confirmation is typically by virus isolation and
identification, or by a specific rise in IgG
antibody, or the presence of IgM antibody.
–isolating virus from the blood during the viremic
phase
 detection of viral RNA (e.g. using polymerase
chain reaction, PCR) or proteins (e.g.,
immunohistochemistry) in frozen or fixed
tissues.
 Alphavirus: Control
 Disease surveillance and virus activity in natural hosts
are used to determine whether control measures will
be undertaken to reduce populations of vector
mosquitoes or to vaccinate hosts, especially horses.
– These measures include control of larvae and adult
mosquitoes
 Human vaccines, where available, are used only in
individuals at particularly high risk of exposure, such
as laboratory workers.
Rubivirus
 Rubivirus
 3 major virion
polypeptides : C and
envelope GP E1 and E2.
 Haemagglutinin (E1)
– E1 is an envelope proteins
responsible for viral
hemagglutination and neutralization
 enveloped,Ss (+) RNA
genome
 replication includes
early (nonstructural) and
late (structural) protein
synthesis
 replicate in the
cytoplasm and bud from
the plasma membrane
Rubella Virus ( German measles )
Latin “ little red”
enveloped virus must stay wet
inactivated by drying, soap, detergents
respiratory virus
does not cause readily detectable
cytopathology
 Rubella virus
Pathogenesis
 Virus infects URT  spreads to local LNviremia 
infection of other tissues and characteristic mild rash
Immune response :
 Penetrates the natural defenses of the nasopharynx and
lung
– Antibody generated blocks viremic spread of the virus
 Viral replication continue until cleared by CMI or limited
by interferon
 Only 1 serotype; Lifelong immunity
 Rubivirinae lack animal reservoirs
 Communicability: 7 days before to 5-7 days after rash onset
- Infants with CRS may shed virus for a year or more
Rubella
mild disease in children and adults, but can
cause devastating problems if it infects the
fetus, especially if infection is in the first few
weeks of pregnancy

Congenital Infection
(+) rubella infection in pregnant women
alter normal growth, mitosis and
chromosomal structure congenital
rubella syndrome (CRS)
Rubella
Clinical syndromes :
I.P. 12-21 days
Children : benign 3 day rash and
swollen glands
Adults : arthralgia, arthritis,
thrombocytopenia, postinfectious
encephalitis
Rubella Clinical Features
 Incubation period 14 days (range 12-23 days)

 Prodrome of low grade fever

 Lymphadenopathy in second week

 Maculopapular rash 14-17 days after exposure

RUBELLA

Murray et al. Medical Microbiology

Congenital Rubella
Syndrome
 severe bilateral
deafness
 severe bilateral
visual defects
 cataract
 corneal opacity
Rubella
Congenital Rubella Syndrome:
triad : abnormalities of the eyes, ears
and heart
eyes : cataract, microophthalmia, glaucoma,
pigmentary retinopathy
ears : bilateral or unilateral sensorineural
deafness
heart : PDA, PA and valvular stenosis,VSD
Rubella
Clinical syndromes :
CNS : microcephaly, psychomotor
retardation, behavioral disorders

persistent infection : progressive

rubella subacute panencephalitis
 Congenital Rubella Syndrome
a) infection of fetus during first trimester of pregnancy
b) at least 20% of infants have severe birth defects
i. neurosensory deafness
ii. blindness (total or partial; cataracts are especially common)
iii. congenital heart disease
iv. microcephaly with mental retardation
c) other symptoms associated with CRS
i. bone translucency and retarded growth
ii. hepatosplenomegaly
iii. Intrauterine growth retardation
d) 10 -20% of babies with CRS die within 1 year
e) 20 % will develop insulin dependent diabetes mellitus as young
adults
f) CRS babies continue to shed Rubella virus from their throats
for several months up to a year after birth and pose a
serious risk to pregnant women.
This infant has the rash of
congenital rubella (German
measles). These infants are at
great risk for severe mental
retardation, deafness, blindness
(atrophic eyes, cataracts and
chorioretinitis) as well as
congenital heart disease and other
abnormalities.

Infant with congenital

rubella and "blueberry
muffin" skin lesions.
 Baby born with rubella:
 Congenital rubella Thickening of the lens of
with hemorrhagic the eye that causes
lesions in the skin blindness (cataracts)
Rubella Complications
Arthralgia or arthritis
children rare
adult female up to 70%

Thrombocytopenic 1/3000 cases

purpura
Encephalitis 1/6,000 cases
Neuritis rare
Orchitis rare
Rubella
Diagnosis :
Clinical
Confirmed : anti rubella specific
IgM
Recent infection : 4 fold rise in specific
IgG titer
Isolation of rubella virus from clinical
specimen (e.g., nasopharynx, urine)
Rubella CDC Case Definition

 Acute onset of generalized

maculopapular rash, and

 Temperature of >37.2 C (>99 F), if

measured, and

 Arthralgia or arthritis, or lymph-

adenopathy, or conjunctivitis
Rubella
Treatment : Supportive

Prevention : live rubella vaccine

Rubella Vaccine
 Composition Live virus (RA 27/3 strain)

 Efficacy 95% (Range, 90%-97%)

 Duration of
Immunity Lifelong

 Schedule >1 Dose

 Should be administered with measles and mumps as MMR

Rubella Vaccine (MMR)
Indications
 All infants >12 months of age

 Susceptible adolescents and adults without

documented evidence of rubella immunity

 Emphasis on non-pregnant women of

childbearing age
 MMR Adverse Reactions
 Fever 5%-15%
 Rash 5%
 Joint symptoms 25%
 Thrombocytopenia <1/30,000 doses
 Parotitis rare
 Deafness rare
 Encephalopathy <1/1,000,000 doses
Rubella Vaccine Arthropathy
 Acute joint symptoms in about 25% of susceptable
adult women

 Frank arthritis occurs in about 10%

 Rare reports of chronic or persistent symptoms

 Population-based studies have not confirmed

association
MMR Vaccine
Contraindications and
Precautions
 Severe allergic reaction to vaccine componen
or following prior dose
 Pregnancy
 Immunosuppression
 Moderate or severe acute illness
 Recent blood product
 Vaccination of Women of
Childbearing Age
 Ask if pregnant or likely to become so in next
4 weeks

 Exclude those who say "yes"

 For others
– Explain theoretical risks
– Vaccinate