HERPESVIRUSES

PRESNTER: FLORA LYIMO

(RESIDENT 2 CMDM)
 Objectives

1. To understand the mechanisms of viral

replication
2. To identify 8 types of Human Herpes Viruses
3. To appreciate clinical manifestations of the
diseases caused by the viruses
 Overview
Important human pathogens

• Ability to establish lifelong persistent infections

• Undergo periodic reactivation
• Some susceptible to antiviral chemotherapy
Classification
Characteristics
 STRUCTURE
• Enveloped viruses
• Icosahedral nucleocapsid
• 100-200 nm in diameter
• Tegument
– Contains proteins and
enzymes
• Double stranded DNA,
the size of the genomes
differs
– Range in length from 120
to 230 kbp
– Base composition from
31% to 75% G+C content
– Contain 60 to 120 genes
Replication
 Transcription
• Transcribed to mRNA by a cellular enzyme
– DNA-dependent RNA polymerase II

• Viral protein synthesis determines whether infection will

result in
– New virus particles and cell death (Lytic infection)
– Persistent shedding of virus
– Latency

• Latency is the property of the host cell

– If do not allow the replication of viral DNA

• Progression beyond the immediate early genes

– Lytic infection will occur
 Translation
• Three classes of proteins need to be made for the production of a
mature virus

• Alpha proteins
– Immediate-early proteins
– Involved in transcriptional regulation
– Involved in the control of beta protein synthesis

• Beta proteins
– Early proteins
– involved in DNA replication
– DNA polymerase and transcription factors

• Gamma proteins
– Late proteins
– structural components of the virus.
– initiated after the start of DNA synthesis
 Replication
• DNA synthesis
– Encode their own DNA-dependent DNA polymerase
– Some encode thymidine kinase that allow growth in non-
dividing cells.
• neurotropic herpes viruses

• Assembly
– Nucleocapsids assemble in the nucleus
– Budd through the double nuclear membrane
– leave the cell via exocytosis pathway
Herpes Simplex Virus
 Herpes simplex viruses
• Two types very similar characteristics

• Genome encodes a number of enzymes

– DNA-dependent DNA polymerase
– Thymidine kinase
– Ribonucleotide reductase

• The genome encodes 11 surface glycoproteins

– Attachment
– Fusion
– Immune escape

• Receptor
– proteoglycan
• heparan sulfate molecule
• Found on the surfaces of many cells
 Pathogenesis
• Retrograde transport to the ganglion
– Trigeminal ganglia
– Sacral ganglia

• Vesicles contains infectious virus

– Heals without scar
– Formation of syncytia
– Cowdry type A inclusion bodies in the nucleus
 Latency
• Only immediate early proteins are made

• Breakage of latency
– virus travels back down the nerve axon

• Recurrence of infection occurs at the same site as the initial infection.

• Triggers of recurrence
– Stress
– Exposure to strong sunlight
– Fever
– Immune suppression

• Recurrent infections, less pronounced than the primary

 Immune Response
• Both cellular and humoral immune response

• Interferon limiting the initial infection

• virus escape the immune system by

– coating itself with IgG via Fc receptors
– Binding complement receptors.
– Cell to cell without entering the extracellular space

• The cell mediated and inflammatory response lead to some of the disease
symptoms
 Epidemiology
• Life-long infection
• Infected can infect others as a result of recurrence

• Antibodies are found in more than 90% of children

– Result of poor hygiene

• Infant infected at birth by a genitally-infected mother

– Severe infection due to undeveloped immunity

• HSV-2 infections are more prevalent later in life

– Number of sexual contacts increases
– The lowest rates found in children
– The highest rates in prostitutes up to 80%
 Treatment
• Variety of nucleoside analog used
– Activated by a viral enzyme, thymidine kinase
– Only activated in herpes-infected cells

• The nucleoside analogs

– Acycloguanosine (acyclovir)

• Act against the replicating virus

– Incorporated into the DNA
Varicella-Zoster Virus
 Varicella-zoster virus
• Causes two major diseases
– Chicken-pox (Varicella)
• Usually in childhood

– Shingles (Zoster) later in life

• Reactivation of an earlier varicella infection
Pathogenesis
Pathogenesis
 Chicken Pox
• Skin rash

• Highly infectious

• 90% of unvaccinated infected in

close contact

• Spread by
– respiratory aerosols
– Direct contact with skin
lesions
Features Small pox Chicken pox

Distribution of Centrifugal Centripetal

Rash
Palms & soles involved Seldom affected

Axilla free Axilla affected

Characteristics Deep seated Superficial

of Rash
Vesicles multilocular Unilocular & dew
& umbilicated drop appearance

Only one stage of Pleomorphic: rash in

rash at one time successive crops

No area of inflammation Area of inflammation

around vesicles around vesicles

Evolution of Slow – macule, papule, Rapid

Rash vesicle, pustule

Scabs form after 10-14 days After 4 - 7 days

 Shingles (Zoster)
• Reactivation under stress or with
immune suppression

• Severe radicular pain

• Chicken pox-like lesions in restricted

areas (dermatome) innervated by a
single ganglion
 Shingles (Zoster)
• Skin lesions
– Maculopapular with an erythematous
base
– Heal in about two weeks

• Post-herpetic neuralgia

• Multi-dermatomal recurrence of
varicella infection
 Diagnosis
• Characteristic appearance
• Definitive diagnosis by culture
• Histology
– characteristic appearance of cells in biopsy specimens of skin
lesions
• Smear of specimen obtained from the base of the lesion (Tzank smear)

• Treatment
• Acyclovir preventing dissemination in mmunosuppressed patients
• Varicella immunoglobulin
• Supportive care in children
– quickly recover if they mount an adequate cell-mediated response
 Prevention
• Live attenuated vaccine
– Leads to antibody production and cell-mediated immunity
– It can be used post-exposure
Epstain – Barr virus
 Epstain – Barr virus
• Causative agent of acute infectious mononucleosis

• Associated with:-
– Nasopharyngeal carcinoma
– Burkitt's lymphoma
– Hodgkin's disease
– Hairy leukoplakia

• There are two types (EBV-1, EBV-2)

– Based on differences in the latency nuclear antigen genes
 Important Properties
• Structurally similar to other, antigenically different

• Viral capsid antigen (VCA)

– Used in diagnostic tests

• Early antigens (EA)

– Produced prior to viral DNA synthesis

• Epstein–Barr nuclear antigen (EBNA)

– located in the nucleus bound to chromosomes

• Two other antigens

– lymphocyte-determined membrane antigen
– viral membrane antigen

• Neutralizing activity is directed against the viral membrane antigen

 Viral Receptors
• Cell types that express the receptor for complement C3d component - CR2 or
CD21
– Epithelial cells (oro- and naso-pharynx)
– B lymphocytes
• Reason for cellular tropism of the virus

• Replication
– Semi-permissive replication
– Permissive replication
 Semi-permissive replication
• B lymphocytes infection may lead to
– latency
– Transformation

• During latency
– Un-integrated copies of virus genome replicated every time the cell
divides.
– Immediate early genes are expressed including the EBNA
– Two latent membrane proteins are expressed
– Two small RNA molecules are expressed
– The membrane proteins are oncogenes
 Permissive replication
• Epithelial cells permit complete lytic replication of the virus

• Epithelial cells allow the expression of proteins

– activates early genes resulting in
• expression of the polymerase and DNA replication
• Capsid proteins
• Membrane glycoproteins
 Epidemiology
• A large proportion of the population (90-95%) is infected
– usually asymptomatic
– Shed the virus from time to time throughout life

• Spread by close contact (kissing disease) and blood transfusion

 Transformation of B cells
• Replicated in pharyngeal epithelial cells

• Shed into the saliva and is taken up by CD21+ B lymphocytes

– Normally short-lived, dying by apoptosis.

• Result of EBV infection,

– B cells are stimulated to divide
– Protected from undergoing apoptosis;
– Becomes transformed and high levels of monocytes are seen in the blood
stream
 Burkitt's lymphoma
• The association between Epstein-Barr virus and Burkitt's lymphoma.

– The tumor cells show evidence of EBV DNA

– Patients show a much higher level of anti-EBV antibodies

– Tumor cells are monoclonal

– EBV can transform B lymphocytes in culture

– EBV can produce B cell lymphomas in primates

 Infectious mononucleosis
• Infected B cells is transformed

• Proliferate and activate suppressor CD8 T cells

• T cells differ from normal

– Downey cells

• T cells increase in number in the circulation up to 80% of the white blood cells.
– results in enlarged lymph glands
– Activation of the T cells limits the proliferation of B cells and the disease
resolves
 Infectious mononucleosis
• The primary infection is often asymptomatic

• Develop infectious mononucleosis after 1-2 months of infection.

• The disease is characterized by

– malaise, lymphadenopathy, tonsillitis, fever, enlarged spleen and liver

• Severity of disease often depends on age

– younger patients resolving more quickly

• Complications
– Meningitis, encephalitis, myelitis and Guillain-Barrè syndrome
– Autoimmune hemolytic anemia, thrombocytopenia,
agranulocytosis, aplastic anemia .
 Serological tests
• Heterophile antibodies are produced by the proliferating B cells
– IgM that interacts with Paul-Bunnell antigen on sheep red blood cells

• No drugs available to treat Epstein-Barr virus

– Reflect the absence of a thymidine kinase encoded by this
virus
Cytomegalovirus (CMV)
 CMV
• Form multinucleated cells (syncytia) with characteristically staining
inclusions

• Macrophages and fibroblasts support a productive infection

• Latent infection is set up in several cell types

– T lymphocytes
– Stromal cells of the bone marrow.

• Only one serotype

 Epidemiology

• Found in significant proportion of the population

• Sero-positivity increases with age

• The virus is spread in most secretions

– Saliva, urine, vaginal secretions, semen
– Semen shows the highest titer of any body fluid

• Infection occurs worldwide

• More than 80% of adults have antibody against the virus

 Transmission
• Variety of modes of transmission

• Early in life
– across the placenta
– During delivery - within the birth canal
– In breast milk

• In young children
– via saliva

• Later in life
– Sexually

• Other mode of transmission

– Blood transfusions
– organ transplants
 Pathogenesis
• First infect the URT then local lymphocytes

• Circulating lymphocytes spread the virus to

– other lymphocytes
– monocytes in spleen
– lymph nodes.

• Spreads to a variety of epithelial cells

– salivary glands, kidney tubules, testes, epididymis and cervix

• Infection is usually asymptomatic (sub-clinical)

– glandular fever in young adults

• Elicits both humoral antibodies and cell-mediated immunity

– Infection is not cleared
– CMI - controls the infection
– Reactivate in cases of immunosuppression
 Clinical manifestation
• Normal person
– Asymptomatic
– Mononucleosis

• Neonate
– Cytomegalic inclusion disease

• Immunosuppresed
– Multisite symptomatic disease
 Congenital disease
• Two instances in which cytomegalovirus can cause serious disease

• Primary infection of pregnant woman, spread via the placenta to the fetus
– cytomegalic inclusion disease
• microcephaly, rash, brain calcification and
hepatosplenomegaly
• hearing loss (bilateral or unilateral) and retardation

• Reactivation in a pregnant woman - in the cervix

– the symptoms are less severe because of the mothers sero-positivity
– congenital abnormalities are rare
 Disease in immunosuppressed
• In patients received an
organ transplant

• Immunosuppressive disease
(e.g. AIDS)
 Diagnosis
• Serology
– fluorescent antibody and ELISA tests
• A fourfold or greater rise in antibody titer

• Histology
– Multinucleated (cytomegalic) cells with characteristic inclusions seen
in biopsies of many tissues

• Cell culture
– Human fibroblasts
– Identified by immunofluorescent antibody

• PCR assays
– Designed to detect replicating virus
– Blood and urine are most commonly tested
• Histology showing nuclear inclusions with
the appearance of an "owl's eye".
 Treatment

• Ganciclovir
– inhibits replication
– Used especially to treat retinitis

• Acyclovir is not effective

 Prevention
• There is no vaccine

• Ganciclovir suppress progressive retinitis in AIDS Patients

• Isolation of Infants with cytomegalic inclusion disease

– shedding virus in urine

• Screening blood for transfusion to newborns

• Screening organs donors for CMV antibody

The best way to avoid the virus is to restrict contact between

infected children and pregnant women
• Transmitted sexually, via saliva, organ transplant
• HHV-8 DNA resembles the lymphotropic herpesviruses
End of presentation

Thank you