Life Cycle of Herpes Simplex Virus a dsDNA GENOME VIRUSES

Greek word herpein ("to creep"), referring to the latent, recurring infections typical of this group of viruses. Herpesviridae can cause latent or lytic infections.

Properties of herpesviruses
Enveloped double stranded DNA viruses.
Genome consisits of long and short fragments which may be orientated in either direction, giving a total of 4 isomers. Set up latent or persistent infection following primary infection

Reactivation are more likely to take place during periods of immunosuppression

Both primary infection and reactivation are likely to be more serious in immunocompromised patients.

Structure
Herpes Simplex Virus (HSV) is a double stranded (ds)DNA virus containing 130-230 Kbp DNA that belongs to Herpesviridae family (Smiley et al. 2004). It contains three main structural components. A central core holds the viral DNA, an inner core is surrounded by an envelope that is made of viral glycoproteins and host cell membranes, and a capsid.

 Between the icosahedral capsid and the lipid envelope is an amorphous protein layer called the tegument. Overall, there are about 40 to 50 different proteins in the virion ("virion structural proteins"). Some of these proteins make up the icosahedral capsid, some make up the tegument, and some are the glycoproteins in the envelope.

Genome
HHV-6 has linear, double stranded DNA which contains an origin of replication, two 810 kb left and right direct repeat termini, and a unique segment that is 143-145kb The HSV genome encodes for over 80 proteins (Khanna et al. 2004). There are three classes of viral genes that are transcribed and translated in a specific order, including the immediate early (IE; ), early (E; ), and late (L; ) genes.
( Immediate early genes (such as -TIF) are transcribed and translated first. Four (ICP0, ICP4, ICP22, and ICP27) of these five proteins from the IE genes serve as regulatory proteins that initiate transcription of early genes by the host cells RNA polymerase (Smiley et al. 2004). ICP4, however, is the main regulatory protein of HSV (Jenkins and Turner 1996). Early genes serve to downregulate immediate early gene expression and upregulate the third set of genes in infection, called late genes. Late genes downregulate early gene expression and are structural proteins (Smiley et al. 2004).)

 After the late phase of infection, a viral capsid is formed in the nucleus that contains viral DNA. The capsid buds through the nuclear membrane and leaves the cell through the Golgi complex. During this process, the virus acquires its tegument and envelop, and the host cell dies as virus is released. HSV is then retrogradely transported along axons to the cell body of neurons to establish a latent infection. This transport is accomplished by dynein and dynactin, which move HSV capsids along microtubules (Dohner et al. 2002).

Epidemiology
HSV is spread by contact, as the virus is shed in saliva, tears, and other secretions.
By far the most common form of infection results from a kiss given to a child or adult from a person shedding the virus.

Primary infection is usually trivial or subclinical in most individuals. It is a disease mainly of very young children ie. those below 5 years.
There are 2 peaks of incidence, the first at 0 - 5 years and the second in the late teens.

Epidemiology
Following primary infection, patients will experience recurrences.
The actual frequency of recurrences varies widely between individuals.

Pathogenesis
During the primary infection, HSV spreads locally and a short-lived viraemia occurs, whereby the virus is disseminated in the body. Spread to the to craniospinal ganglia occurs. The virus then establishes latency in the craniospinal ganglia.

The exact mechanism of latency is not known, it may be true latency where there is no viral replication or viral persistence where there is a low level of viral replication.
Reactivation - It is well known that many triggers can provoke a recurrence. These include physical or psychological stress, infection; especially pneumococcal and meningococcal, fever, irradiation; including sunlight, and menstruation.

Laboratory Diagnosis
Direct Detection
Electron microscopy of vesicle fluid PCR - now used routinely for the diagnosis of herpes

Indirect or Serology
Not that useful in the acute phase because it takes 1-2 weeks for before antibodies appear after infection. Used to document to recent infection.

Cytopathic Effect of HSV in cell culture: Note the ballooning of cells. (Linda Stannard, University of Cape Town, S.A.)

Positive immunofluorescence test for HSV antigen in epithelial cell. (Virology Laboratory, New-Yale Haven Hospital)

Prevention
The virus is ubiquitous nothing can be done to prevent the transmission of infection in environmental terms. Prophylactic chemotherapy may be given to those suffering from frequent and severe recurrent herpes. In clinical trials, a -interferon eye drops have proved effective for the prevention of recurrent dendritic ulcers.

Management
At present, there are only a few indications of antiviral chemotherapy, with the high cost of antiviral drugs being a main consideration. Acyclovir this the drug of choice for most situations at present. Famciclovir oral only, more expensive than acyclovir.

HSV_vaccines
Several recombinant subunit vaccines are being evaluated at present. There is evidence to suggest that such vaccines may be effective in reducing the frequency and severity of recurrent disease in an already immune individual, but their efficacy in preventing primary infection is uncertain.

The replication cycle of Herpes Simplex virus

1. Specific proteins in the viral envelope attach to host cell receptors on the cell membrane. 2. Penetration is achieved when the viral envelope fuses with the cell membrane releasing the nucleocapsid directly into the cytoplasm. 3. The virion is uncoated and the viral DNA is transported into the nucleus. 4. In the nucleus, the viral DNA is transcribed into early mRNAs which are transported to the cytoplasm for the translation of early proteins. These early proteins are brought back into the nucleus and participate in the replication of the virus DNA into many copies.

 The viral DNA is then transcribed into the late mRNAs which exit to the cytoplasm for translation into the late (nucleocapsid and envelope) proteins. 5. The capsid proteins encapsidate the newly replicated genomes. The envelope proteins are imbedded in the nuclear membrane. 6. The nucleocapsids are enveloped by budding through the nuclear membrane, and the mature viruses are released from the cell through cytoplasmic channels.

Primary Infection: The Lytic Cycle

HSV infects its host through both lytic and latent infection, and replication of HSV occurs within 15 hours after infection (Jenkins and Turner 1996). In the lytic cycle, HSV infects epithelial cells located in the mucosa, replicates, and causes epithelial cell death . In order to infect epithelial cells, glycoproteins (namely gB, gC, and gD) on the surface of HSV fuse with entry receptors on the host cell membrane.

 There are three known types of entry receptors located on the host cell that bind to HSV during cell infection and fusion. Heparan sulfate is a glycosaminoglycan (GAG), and it binds to gB and gC (Spear et al. 2004).

The sequence of events at the cell surface usually involves the HSV-1 virion binding initially to heparan sulphate, and then to the main receptor. The latter can be one of several types of cell surface molecule including some nectins, which are cell adhesion molecules. The virion envelope then fuses with the plasma membrane. Infection may also occur by endocytosis, followed by fusion between the virion envelope and the endosome membrane.

Adenovirus

 Adenoviruses are medium-sized (90 100 nm), nonenveloped (without an outer lipid bilayer)icosahedral viruses composed of a nucleocapsid and a double-stranded linear DNA genome. The virus is composed of around 1 million amino acid residues and weighs around 150 MDa

Structure
Adenoviruses represent the largest nonenveloped viruses. Because of their large size, they are able to be transported through the endosome (i.e., envelope fusion is not necessary). The virion also has a unique "spike" or fiber associated with each penton base of the capsid that aids in attachment to the host cell.

Disease
Adenoviruses can infect the respiratory tract; eye, gastrointestinal tract and bladder. (CNS infection has been documented but is very rare). In young children and infants it causes an acute febrile upper respiratory tract infection. It may progress to pneumonia and pharyngeal infection in untreated cases and in immuno-compromised individuals. In adults the symptoms include fever with pneumonia and pharyngitis. 5% of acute respiratory disease in children under 5 years of age is caused by adenoviruses and 10% of the pneumonias. Ad. infections are difficult to distinguish from influenza, parainfluenza and RSV. Conjunctivitis can occur with respiratory illness in such cases the disease is called pharyngoconjunctival fever.

Epidimiology
Adenoviruses are highly species specific. Fecal oral transmission is common in children.

Diagnosis
Adenovirus infections can be identified using antigen detection, polymerase chain reaction assay, virus isolation, and serology.

Prevention
Good hygiene, including hand washing, is still the best way to avoid picking up the adenovirus from an infected person. In the past, US military recruits were vaccinated against two serotypes of adenotypes, with a corresponding decrease in illnesses caused by those serotypes. The vaccine is no longer manufactured, and there are currently no vaccines available to protect against the adenovirus.

Treatment
There are no antiviral drugs to treat adenoviral infections, so treatment is largely directed at the symptoms.

Genome
The adenovirus genome is linear, nonsegmented double-stranded (ds) DNA that is between 26 and 45 Kbp. This allows the virus to theoretically carry 22 to 40 genes.

 Entry of adenoviruses into the host cell involves two sets of interactions between the virus and the host cell. Most of the action occurs at the vertices. Entry into the host cell is initiated by the knob domain of the fiber protein binding to the cell receptor. The two currently established receptors are: CD46 for the group B human adenovirus serotypes and the coxsackievirus adenovirus receptor (CAR) for all other serotypes.

 To enter the cells they use a receptor present in the host cell called as CAR (coxsackie and adenovirus receptor). The internalization of the virus particles occur through receptor mediated endocytosis. After entry the endosome containing the virus particle migrates to nucleus and the genetic information of the virus is released into the nucleus. Transcription of the first gene is done by the terminal protein attached with the viral DNA. Viral mRNA is then transported to the cytoplasm and translated into the viral proteins. Virus assembly takes place in the cytoplasm and the mature viral particles get released from the infected cells after killing them by accumulated adenoviral death proteins.

Replicative Cycle
The adenovirus replicative cycle is divided into early and late phases with the late phase occurring when viral DNA replication begins. Attachment-Penetration-Uncoating Adenovirus attachment is mediated by the fiber which binds a specific receptor on the cell membrane. Subsequently the attached virus migrates to clathrin coated pits to form a receptosome and become internalized.

 Early Transcription Viral gene expression is not a haphazard process. On the contrary it is highly ordered and carefully orchestrated in much the same manner as genes are regulated for cell division or embryonic development.

SV40

 SV40 is an abbreviation for Simian vacuolating virus 40 or Simian virus 40, a polyomavirus that is found in both monkeys and humans. Like other polyomaviruses, SV40 is a DNA virus that has the potential toSV40 is an abbreviation for Simian vacuolating virus 40 or Simian virus 40, apolyomavirus that is found in both monkeys and humans. Like other polyomaviruses, SV40 is a DNA virus that has the potential to cause tumors, but most often persists as a latent infection. cause tumors, but most often persists as a latent infection.