Who is this

Togaviridae?
Since its inception, the members of the virus family Togaviridae have been anything but constant. Classified originally on the basis of being small nonsegmented positive sense single strand RNA viruses the classification was anything but precise. Dubbed Togavirus because of the "loose envelope" displayed in electron micrographs, the togaviridae were later shown to have envelopes that are quite tightly bound to the nucleocapsid. So general was the classification criteria of Togavirus, that three virus divisions were discovered within it to be very different from each other. The family Togaviridae was then split into three in 1984, joined by its cousins Flaviviridae and Pestiviridae (often not considered a true viral family). The split was a result of advances in microbiology techniques, and especially the development of powerful genetic analyzation techniques. It was realized that the three groups went about replication in strikingly different ways, and the distinction between the families was established. A major unifying principle between all three groups is that they are all capable of being transmitted by arthropod vectors. The proximity between Togaviridae and Flaviviridae is maintained by their respective vector-borne groups, Alphavirinae and Betavirinae. Surprisingly, Togaviridae is less known for its arthrpod borne infectivity than for the single member of the Rubivirinae, Rubella. Originally recognized as the "German Measles" in th early nineteenth century, the virus was disregarded as a mild infection of little importance. In an age where small pox, influenza, and pneumonia were

the leading killers of man, it is not hard to believe that a virus with an a mortality rate of 0.05% would be overlooked. Rubella outbreaks were common in the spring across the United States with a periodicity of about 3 years. 80% of americans were seropositive for rubellaby childbearing age. For over a century this highly contagious virus was overlooked, until an astute opthamologist took notice of it. In 1941, Norman Gregg began to note an unusually high incidence of congenital cataracts. After some research, he found that the cataracts were closely related to a maternal Rubella infection in the first trimester of pregnancy. This was one of the first demonstrations of a virus as a teratogen. Some follow-up studies later showed that firsttrimester maternal infections of Rubella were also correlated with more severe birth defects such as deafness and blindness, and also with miscarrage. Children recognized to have congenital abnormalities due to prenatal infection are termed to have congenital rubella syndrome, or CRS. Public response to the discoverery of a viral teratogen was marked. Parents forced children to attend "rubella parties" with people known to be infected with rubella modelled after "measles parties" intended to ensure children were exposed to the disease and developed immunity at an age where the incidence serious sequella are reduced. Because rubella has only one serotype, lifelong immunity is acquired after the initial immune response. Rubella virus was isolated in 1962, and a live, attenuated vaccine was immediately produced. 1964 marked the last epidemic of Rubella observed in the United States. In 1972 the vaccine was merged with the measles and mumps vaccine to become the MMR vaccine, commonly given to children at the age of 15 months. The alphavirinae, though far greater in number, and equally, if not more, pathogenic than the rubivirinae, have taken the back seat to rubella. The alphaviruses are transmitted only by arthropod vector bite. Although the viruses replicate in man, the viruses do not become concentrated enough to be transmitted. As a result, alphavirinae rarely reach epidemic proportion, and only as a result of extreme proximity of man and insect population. Most commonly transmitted by various mosquito species, the alphavirinae are capable of replication within the vector. Animal hosts include horses and birds, in which the viruses are mildly pathogenic, presenting little more morbidity than rubella in adult man. The alphavirinae present two distinct types of disease, one type involves encephalitis and is highly linked with horses, the other is characterized by arthalgia (arthritic pain) and mild fever. Alphavirinae are present

throughout the world, and arise periodically in the spring when conditions for viral replication are ideal and vector populations are large. Vaccines have been developed against the encephalitic deiseases, which are characterized by fatality rates of 30-70%. Human Togaviruses and their diseases

Alphaviruses
Encephalitic Viruses Eastern Equine Encephalitis (EEEV)- Originally native to Japan, it was spread to North America in shipments of tires containing the viral vector Aedes Albopictus, an eastern Tiger mosquito. Symptoms appear 5 to 15 days after being bitten by an infected mosquito, include high fever, headache, stiff neck and myalgia. One in three infected individuals will come down with the serious disease, two in three will be able to fight off the viral infection. The disease is fatal in 30-50% of cases Western Equine Encephalitis (WEEV)- Most common of the Equine Encephalitises in the Midwest and Western portions of the United States. Transmitted via mosquito vector. Cases have been reported in Western USA, West Central Canada, Mexico and South America. Symptoms similar to EEE. Venezuelan Equine Encephalitis (VEEV)- Arboviral disease that represents a threat throughout most of the Americas, including the United States. This virus has caused repeated epidemics and equine epizootics since the 1920s, involving up to hundreds of thousand of equines and tens of thousands of people with severe morbidity and mortality. Symptoms similar to EEE. Everglades virus (EVEV)- A variation of VEEV. Antigenic and animal virulence studies show distinct differences between the endemic Florida strain, which has never been seen to cause overt disease in equines or humans, and the more virulent epizootic strains of VEE found in Central and South America led to the classification of the Florida strain of VEE as a separate Group A arbovirus now known as Everglades virus. Arthritic VirusesChikungunya (CHIKV)- characterized by sudden onset, chills and fever, and intense arthritic pain. The illness usually lasts 3-7

days. The word "chikungunya" is Swahili for "that which bends up," in reference to the stooped posture of patients afflicted with the severe joint pain associated with this disease. Chikungunya virus has been isolated from humans and mosquitoes in eastern, southern, western, and central Africa and in southeastern Asia, where it has been responsible for illnesses in hundreds to thousands of individuals. O'nyong-nyong (ONNV)- Transmitted by Anopheles funestus and An. gambiae mosquitos . Following an incubation period of three to twelve days, the patient experiences myalgia and severe arthritis. Patients recover after about two weeks. 'O'nyong nyong' means 'weakening of the joints' in the Acholi dialect. The virus was first isolated during an epidemic in Acholi, Uganda. Several million subsequent cases were reported in Uganda, Kenya, Tanzania, Malawi and Mozambique during 1959 to 1961. Ross River (RRV)- Causes the disease commonly know as epidemic polyarthritis (EPA), was first isolated in 1963. The disease is most common in eastern Australia though it has been diagnosed throughout Australia. Ross River Virus is characterised by arthritic symptoms. Barmah Forrest (BFV)- Generally characterised by arthritic symptoms, similar to that of Ross River virus. Cases of Barmah Forest virus have been diagnosed throughout eastern Australia, especially in the northern state of Queensland. Mucambo virus (MUCV)Sindbis (SINV)- Characterized by coincident onset of fever with rash and arthritic pain. Sindbis is found in Africa, India, and Malasia, and has been observed to reach epidemic proportions in Egypt and South Africa. Mayaro- Also characterized by rash, fever, and arthritic pain, Mayaro is found in Central and South America. Many epidemics have been reported in the Amazon Basin. Semliki Forest Virus (SFV)-

Rubiviruses
Rubella (RUBV)- A highly contagious mild infection of early childhood. Often refered to as the "German Measles", due to the

diffuse maculopapular rash and fever it causes. When infection occurs during pregnancy, especially during the first trimester the virus is extermely teratogenic. Fetal infection is likely and often causes congenital rubella syndrome (CRS) resulting in abortions, miscarriages, stillbirths, and severe birth defects. Up to 20% of the infants born to mothers infected during the first half of pregnancy have CRS. The most common congenital defects are cataracts, heart disease, sensorineural deafness, and mental retardation.

Classification and Taxonomy

GenomeMonopartite, positive sense, single stranded RNA. 5' capped, 3' polyadenylate d 10-12 kb long Cytoplasmic replication Nonstructural proteins encoded at the 5' end
Schematics by Steve Folder

Capsid-

Envelop ed T=3 icosahe dral nucleoc apsid 40-70 nm diamete r Compri sed of multime From Thomas Smith's alphavirus page rs of 1 gene product Spheric al

EnvelopeTightly bound to capsid 2 peplomers: E1 and E2 are responsible for viral specificity Envelope acquired by budding

Translation & Protein ManipulationBecause RNA is positive sense, it is immediately able to be translated by cellular ribosomes and tRNA. Two-thirds of the RNA on the 5' side are translated into a polyprotein, which subsequently cleaves itself into four nonstructural proteins (NSP). NSP1- methylation and capping of viral RNA NSP2- helicase and protease NSP3- converts RNA replicase to plus strand replicase

NSP4- replicase Following translation of the non-structural viral proteins, the positive sense genome is transcribed tomproduce a negative sense copy. From this negative sense copy, a full length positive sense genome is produced for packaging, and a sub-genomic 3' mRNA fragment encoding the structiral proteins is made. The remaining 3' third of viral RNA is translated into another polyprotein that encodes all of the stuctural proteins. Alphavirinae's polyprotein is autolytic for the C protein, which is cleaved while the protein is in the cytosol. The remaining envelope proteins insert into the endoplasmic reticulum, where they are cleaved by host-cell signalase. Following glycosylation the envelope proteins are released back into the cytosol, where they bind to the cellular membrane. Rubivirinae goes about cleavage in a slightly different manner by initially cleaving the E1 protein from E2 in the cytosol, The E1 and transmembrane proteins are cleaved by host-cell signalase in the endoplasmic reticulum. The E2-Capsid protein is cleaved by host-cell signalase and immediately associates with the membrane for viral assembly. The difference in management and manipulation of the NSP polyprotein, and gene order, in addition to vector trasferability, are the major criteria used to separate the alphavirinae from the rubivirinae. C- capsid E1, E2, (E3)- Envelope peplomer Small transmembrane protein The nucleocapsid protein spontaneously assembles with inserted envelope proteins to form a tightly-bound nucleocapsid-envelope complex. This complex then selectively packages genomic RNA over the sub-genomic mRNA encoding the NSP polyprotein. This phenomenon has been observed in in vivo studies, but has not been repeated in vitro, suggesting that the mechanism of genome uptake is not fully understood. The virus acquires its envelope by budding through the cellular membrane. Alphavirinae, which bud through cytoplasmic membranes, are further distinguished from the rubivirinae, which bud through intracytoplasmic membranes.

Image is public domain

Infection Cycle
The infection cycle of togaviridae begins when the virion is taken up into cellular vessicles by receptor-mediated endocytosis. A pH drop caused by the merging of the carrier vessicle with a cellular lysosome digests nucleocapsid proteins and releases the genomic RNA to the cytosol where replication begins. The positive sense RNA is immediately translated as describes above. New virions spontaneously assemble following viral translation and replication. The virion then buds through cellular membranes. Alphavirinae are considered to be zoonoses of man, as their main host are domesticated and wild animals. Parasitized by mosquitos, the virus is taken up in the animal host's blood. The virus replicates within the mosquito vector before it is introduced into a human host through mosquito bite. The virus is introduced into the capilarries, and replicates in the vasular endothelium

before being spread through the blood to other target tissues including muscles, joints, skin, and brain.

Image is public domain

Rubivirinae lack animal reservoirs, and are spread between man easily. The the virus is spread via aerosol, and enters the body trough inhalation. It establishes infection in the upper respiratory tract, and spread to the lymph nodes. The virus incubates for ~18 days and manifests itself as a rash with fever.

Alphavirus
Species: Aura virus Species: Barmah Forest virus Species: Bebaru virus Species: Cabassou virus Species: Chikungunya virus Species: Eastern equine encephalitis virus Species: Everglades virus Species: Fort Morgan virus Species: Getah virus Species: Highlands J virus Species: Mayaro virus

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Species: Middelburg virus Species: Mosso das Pedras virus (78V3531) Species: Mucambo virus Species: Ndumu virus Species: O'nyong-nyong virus Species: Pixuna virus Species: Rio Negro virus Species: Ross River virus Species: Salmon pancreas disease virus Species: Semliki Forest virus Species: Sindbis virus Species: Southern elephant seal virus Species: Tonate virus Species: Trocara virus Species: Una virus Species: Venezuelan equine encephalitis virus Species: Western equine encephalitis virus Species: Whataroa virus