Introduction to DNA Viruses II 1. Describe the structure of herpes virus. Herpesvirus is large! is enveloped.

d. Contains numerous glycoproteins has a tegument. This is a protein-filled amorphous-looking region that lies between the envelope and capsid. has an icosahedral capsid. This surrounds the core. has a toroidal-shaped proteinaceous core that is surrounded by the large dsDNA genome. The large dsDNA genome has inverted repeats that allow for viral DNA circularization. It also contains multiple repeated sequences and varies greatly in size (by up to 10kbp). Infection by Herpesvirus is virtually universal in the human population. Most adults test seropositive. There are eight known Herpesviruses divided into three sub-families: alpha: HSV1, HSV2, and VZV (varicella zoster) beta: CMV, herpes virus 6, and herpes virus 7 gamma: EBV, Kaposi sarcoma virus (aka HHV8) 2. Understand the life cycle of herpes virus.
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Adsorption/attachment: process is mediated by envelope glycoproteins (there are 9 of them). Cellular receptors are not known, but HSV binds to different receptors on different cell types (maybe tissue tropism?). Entry: the glycoprotein interactions with cellular receptors result in direct fusion of the envelope with the cell membrane, involving uncoating at the plasma membrane. Fusion releases the capsid into the cytoplasm, where it migrates to the nucleus. The core then enters the nucleus through nuclear pores. Genome circulariziation: via inverted repeats in the dsDNA genome. Gene expression: transcription occurs via cellular RNA polymerase II but is regulated by both viral and host transcription factors. Gene expression is divided into three temporal phases: a. Alpha phase: immediate early phase that primes the cell for further viral gene expression and recruitment of transcriptional machinery b. Beta phase: early phase that involves expression of genes that are directly involved in viral genome replication c. Gamma phase: late phase that involves the expression of viral structural proteins DNA replication: translation is carried out by viral DNA polymerase in a complex pattern that involves at least three potential origins of replication. a. Replication begins with bi-directional replication followed by a switch to a rolling circle b. The replication process results in the formation of high molecular weight DNA concatemers

Late gene expression: this is the aforementioned gamma phase of DNA replication, when structural proteins are produced 7. Virus assembly: the high molecular weight DNA concatemers are cleaved and packaged into pre-assembled capsids in the nucleus. The viral capsids then move from the nucleus into the cytoplasm, where they are surrounded by tegument proteins and bud into exocytotic vesicles that migrate towards virus-modified cell membrane. 8. Virus exit: nascent virus fuses with cell membrane regions that contain the viral glycoproteins. They are released from the cell.
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3. Understand the concept of latency. During Herpesvirus primary infection, the virus entersa latent (silent) phase of infection that allows the virus to escape immune response by hiding in the following cell reservoirs: HSV and VZV hide in neuronal roots of ganglia EBV and HHV8 hide in B lymphcytes CMV hides in T cells and macrophages In these reservoirs, the viral DNA is maintained as an episome and therefore not integrated. Hence, the viral genome is mostly not expressed, with the exception of limited regions of specific genes called latency associated transcripts (LAT) that help maintain latency. How does Herpesvirus exit latency (get reactivated)? Exposure to UV light (i.e. sunshine!) Suppression of immune system Fever Trauma to latently infected nerve cells Pneumococcal pneumonia stress 4. Describe the diseases associated with herpes virus infection. HSV1 and HSV2 cause oral herpes (cold sores) and genital herpes how: infection of epithelial cells of mucous membranes of the mouth, noses, eyes, and genital tract, or entry of the body through wounds; entry is via wounds/breaks in mucous membranes course of infection: initial infection causes reddened area that blisters and crusts over; blister contains fluid full of infectious virus! Virus travels up nerve cells and establishes itself in the nerve ganglion where it becomes latent Periodic reactivation causes the virus to travel back down nerve cells to the initial site of infection, resulting in recurrent outbreaks EBV causes heterophile-positive infectious mononucleosis (glandular fever) and is associated with Burkitts Lymphoma, Hodgkins lymphoma, nasopharyngeal cancer, and oral hairy leukoplakia (oral lesions that are an opportunistic infection in HIV-infected patients) How: infection of epithelial cells of mucous membranes in the throat and B lymphocytes Course of infection:

Viral replication occurs in the mucous membranes and is shed in saliva Viruses in the saliva are taken up by B cells, which proliferate resulting in the production of antibodies (hence the name heterophile positive mononucleosis) Virus remains latent in the B cells Latent EBV is implicated in Hodgkins lymphoma and nasopharyngeal carcinoma Burkitts is most commonly seen in AIDS patients with EBV associated infection Infection of T cells causes increase in atypical lymphocytes called Downey cells Infection of B and T cells causes swelling of lymph nodes EBV has also been implicated in lymphoproliferative disorders in the immunocompromised (i.e. transplant patients) as well as individuals with congenital T cell defects (i.e. X-linked lymphoproliferative disease) VZV causes chicken pox (Varicella) in children and shingles (zoster) later in life How: infection of epithelial cells of respiratory mucous membranes Course of infection: Viral spread from lungs to lymphocytes, monocytes, and reticuloendothelial system Subsequently spreads to skin, mouth, conjunctiva, respiratory tract, and skin Rash develops on face, scalp, trunk, and less often on arms and legs Transmission: spread through contact, conjunctivitis, or respiratory route can also be transmitted sexually, vertically from infected mother to fetus, from infected mother to newborn via breastfeeding, from blood transfusion, and transplantations. Zoster is a consequence of reactivation of chicken box infection CMV causes heterophile-negative mononucleosis syndrome and congenital cytomegalic inclusion disease (CID) How: infection of epithelial cells and fibroblasts Course of infection: CMV establishes latent infection in mononuclear leukocytes and in organs such as kidney and heart Infection causes enlargement of cells and nuclear inclusion bodies in a wide range of tissues (systemic infection) Cell-mediated immunity is critical in controlling and resolving CMV growth; consequently, complications from CMV infection are most commonly observed in neonates and patients with T cell defects (i.e. AIDS and transplant patients*) CMV ends up stimulating T cell growth without the accompanying antibody production (hence the name heterophile negative mononucleosis) CMV causes mild pharyngitis that is a less severe form of mono than that caused by EBV Fetal congenital infections are a consequence of a pregnant mother being infected with CMV, resulting in congenital cytomegalic inclusion disease that can result in microcephaly, intracerebral calcification, rash, deafness, and mental retardation

*Immunocompromised (transplant or AIDS patients) undergo reactivation of the virus due their weaker cell-mediated immune systems; consequences include retinitis, pneumonia, and GI infections (gastritis, colitis, esophagitis, hepatitis) Transmission: contact with body fluids such as saliva, blood, semen, and tears; can also be spread via transplantation HHV 6 causes exanthum subitum (roeolum infantum) in young children How: virus infects and replicates within T and B cells, megakaryocytes, glioblastoma cells, and the mouth and throat Course of infection: Fever, and sometimes respiratory tract infection and swelling of lymph nodes, is subsequently followed by rash (exanthema!) on the neck and trunk Latent infection continues in T cells HHV7 causes exanthum subitum similar to HHV6 How: HHV7 is found in the saliva of most adults Replicates and becomes latent in T cells HHV8 causes Kaposis sarcoma (consequently, HHV8 is sometimes called Kaposis sarcomaassociated virus) How: HHV8 infects peripheral lymphocytes It is found in the saliva of many HIV positive patients

Note on antivirals: Nucleoside analogs are highly selective drugs against the Herpesvirus. They include: Acyclovir: the drug is phosphorylated by HSV thymidine kinase into an active form that inhibits viral DNA polymerase Primarily used to treat VZV and HSVI and II Gancyclovir: specifically phosphorylated by CMV-encoded kinase Consequently, gancyclovir is used particularly for CMV