HERPES VIRUS

INTRODUCTION:
Herpes virus are included in the family Herpesviridae, which have been divided into three
sub-families Alphaherpesvirinae, Betaherpesvirinae, Gammaherpesvirinae. Containing over
a hundred species of envelope DNA virus, characterised by their ability to establish life-long
latent infections which enable them to undergo periodic reactivation.

MORPHOLOGY:
Herpesvirus is 100-200 nm in diameter, the envelope virion measures about 200 nm while
the naked virion is about 100 nm in diameter, which contains icosahedral (Geometric shape
with 20 sides) capsid which are composed of 162 capsomeres enclosing linear double
stranded DNA genome and is surrounded by lipid envelope containing peplomeres (spike
proteins) about 8 nm long derived from modified host nuclear membrane through the
naked virions bud during replication. Between capsid and envelope is an amorphous
structure called ‘tegument’ which contains several proteins.

Fig: Herpes simplex virus

 Fig :Electron micrograph of HSV

Replication & Resistance:

Herpesvirus multiply in the nuclei of infected cells and produce Cowdry type A intranuclear
inclusion bodies
Like other envelop virus they are susceptible to fat solvents like alcohol, ether,
chloroform & bile salts.
Classification:
Eight different types of Herpesvirus are known whose primary host are human, they are
divided into 3 subfamilies based on biological, physical & genetic properties:
• Alphaherpesvirinae with a relatively short replicative cycle(12-18 hrs)
• Betaherpesvirinae which replicates slowly less than 24 hrs
• Gammaherpesvirinae

SUBFAMILY SCIENTIFIC NAME COMMON NAME

Alphaherpesvirinae Human herpesvirus 1 Herpes simplex virus type 1

Human herpesvirus 2 Herpes simplex virus type 2
Human herpesvirus 3 Varicella-zoster virus
Cercopithecine herpesvirus 1 Herpes B virus
Betaherpesvirinae Human herpes virus 5 Cytomegalovirus
Human herpes virus 6a -
Human herpes virus 6b -
Human herpes virus 7 -
Gammaherpesvirinae Human herpes virus 4 Epstein-Barr (EB) virus
Human herpes virus 8 Kaposi’s sarcoma associated virus

The herpes virus family has no common group antigen & the different species do not show
any significant antigenic cross reaction, except between herpes simplex type 1 and 2.
 HERPES SIMPLEX VIRUS
INTRODUCTION:
The herpes simplex virus (HSV) occurs naturally only in humans, but it can produce
experimental infection in many laboratory animals. The virus is of two types -- HSV Type 1
(Human herpes virus type 1) which is usually associated with oral and ocular (related with
eye) lesion and is transmitted by direct contact or droplets spread from cases or carriers.
HSV Type 2 (Human herpes virus type 2) is responsible for majority of genital
infection, it is commonly transmitted venerally (by sexual means).

CULTURAL CHARACTERISTICS:
The virus grows in a variety of primary and continuous cell culture (monkey or rabbit
kidney, human amnion, HeLa) producing cytopathic change, well defined foci with heaped
up cells and syncytial or giant cell formation on chick embryo chorioallantoic membrane
(CAM), small (diameter less than 0.5 mm, white shiny non necrotic pocks are produced).
Inoculation in mice and on chick embryo CAM is not sensitive and has been replaced by
tissue culture for virus isolation. Primary human embryonic kidney, human amnion and
many other cells are susceptible, but human diploid fibroblast are preferred. Typical
cytopathic changes may appear as early as in 24- 48 hrs, but culture should be observed for
2 weeks before declared negative. Drug susceptibility too can be tested in cell cultures. The
two types of virus cross-react serologically.
Differentiating features:
• On chick embryo CAM, type 2 strains form larger pocks resembling variola.
• Type 2 strain replicate well in chick embryo fibroblast cells, while type 1 strains do so
poorly.
• Type 2 strains are more neurovirulent in laboratory animals than type 1.
• The infectivity of type 2 is more temperature sensitive than that of type 1.
• Type 2 strains are more resistant to antiviral agents like IUDR and cytarabine in
culture.
• Restriction endonuclease analysis of viral DNA enables differentiation between the
two types as well as between strains within the same type.

Fig: Chick embryo CAM HSV 1&2, 5 days old culture

a ) Large pocks produce by HSV-2 b ) Pocks size remain same in case of HSV-1
Fig: a) Vervet monkey kidney cells (Vero cell line) unaffected
b) Vervet monkey kidney cells (Vero cell line) infected with HSV (rounded cell & giant cell
formation).

PATHOGENICITY:
Herpes simplex is one of the most common viral infections in humans, about 60-90% of
adults showing detectable antibody. Primary infections occur during childhood (about 5
yrs), humans are the only natural host and the sources of infection are saliva, skin lesions or
respiratory secretion. Asymtomatic carriers are the most important sources of infection,
especially in genital herpes.

ROUTE OF INFECTION: The disease is transmitted by close contact and may be sexually
transmitted in genital herpes. The virus enter through the defect in the skin or mucous
membrane and multiply locally resulting in a vesicle formation. The virus spreading to
draining lymph nodes producing lymphadenitis.
After primary infection the virus travel by retrograde intraaxonal flow to sensory
root ganglia. They settle within the neurons in ganglia most common being trigeminal(HSV-
1) and sacral(HSV-2) ganglia. The herpes virus DNA get integrated into the host cell genome
and get reactivated by stimuli such as common cold, pneumonia, stress, fever, exposure to
sunlight etc. Antibodies may not prevent recurrences, but can reduce the severity of the
clinical diseases.

CLINICAL FEATURES:
The clinical manifestations and course depend on the site of infection, age and immune
status of the host, and the antigenic type of the virus.

Cutaneous infection: Most common site on the cheeks, chin and around the mouth or
forehead, lesion may also appear in infants as napkin rash.The typical lesion is the ‘fever
blister’ or herpes febrilis caused by viral reactivation in febrile patients.
Mucosal: Buccal mucosa is the most commonly affected site. Gingivostomatitis and
pharyngitis are the most frequent conditions in primary and herps labialis in recurrent
infections.

Ophthalmic: HSV is the most common cause of corneal blindness in developed countries.
Acute keratoconjuctivitis may occur by itself or by extension from facial herpes. follicular
conjunctivitis with vesicles formation on the lid is another manifestation.

Nervous System: HSV encephalitis, though rare, is the most common sporadiac acute viral
encephalitis in most parts of the world having an acute onset, with fever and focal
neurological symptoms.

Genital Herpes: Categorised as veneral diseases,in men the lesions occur mainly ion the
penis and or in the urethra causing urethritis. In women, the cervix, vagina, vulva and
perineum are affected.

Congenital: Transplacental infection with HSV 1 or 2 can lead to congenital malformations

but this is rare.Infections may occur during birth, particularly if the mother has genital
lesions due to HSV 2.

Infection cause by HSV 1 and HSV 2

HSV 1
-acute gingivostomatitis
-pharyngotonsilitis
-herpes labialis
-keratoconjunctivitis
-eczema herpeticum
-encephalitis
-dendritic keratitis
HSV 2
-genital herpes (penis, urethra, cervix, vulva, vagina)
-neonatal herpes
-aseptic meningitis

Fig: Fever blister

 Cutaneous lesions in herpes occurs generally on ‘skin above the waist’in HSV-1 and ‘skin
below waist’in HSV-2.Herpetic whitlow is an occupational hazard of health workers,who
acquire infection from saliva and respiratory secretion of patients.Vesicles may be
produced on skin.
Beside above mention primary infections, herpes virus may be present as latent
infection, reactivation and recrudescence
Characterictics HSV-1 HSV-2
A.Typical transmission Contact (Often saliva) Sexual
B. Infections
- Acute gingivostomatitis + -
- Pharyngotonsilitis + -
- Kerato Conjuctivitis + -
- Neonatal herpes ± +
- Cutaneous herpes
Skin above the waist + ±
Skin below the waist ± +
- Herpetic whitlow + +
- Eczema herpeticum + -
- Genital herpes ± +
- Encephalitis + -
- Asceptic meningitis - +
C.Latency
1.Typical site Trigeminal ganglia Sacral ganglia

LABORATORY DIAGNOSIS
1.Specimens:
Specimens include vesicle fluid ,skin swab, saliva, corneal scraping, brain biopsy and CSF,
according to the site of involvement.

2.Direct Examination:
(i) Smears prepared from scrapings from the base of vesicles are stained with
toluidine blue. Multinucleated giant cells with faceted nuclei and homogeneously stained
‘ground glass’ chromatin(Tzanck cells) are present in positive smear.

(ii)Cowdry type A intranuclear inclusion bodies may be seen Giemsa stained smears.

(iii)Herpes virions may be demonstrated in specimens by electron microscopy.

(iv)Viral antigens can also be demonstrated in the scrapings from the base of lesions and
tissue preparations, stained by immunofluorescence staining .The fluorescent antibody
test on brain biopsy specimens provide reliable and early diagnosis in case of encephalitis.
 3.Tissue Culture:
Virus can be isolated on human fibroblasts, HEp-2 cells, Vero cells and chorioallantoic
membrane. Swollen, rounded cells may appear within 1-5 days. Some virus
strains(particularly HSV-2 strains)may give rise to syncytium(fusion of infected cells)
formation. Diagnosis can be confirmed by immunofluorescent staining of infected cell
culture.HSV-1 and HSV-2 can be differentiated by abuse of monoclonal antibodies in
immunofluorescent staining or by neutralisation test with specific antiserum.

4.Serology:
Primary infections can be diagnosed by detection of virus specific IgM antibody or by a
rising titre of antibody. Various test like compliment fixation test(CFT), neutralisation,
immunofluorescence, ELISA and RIA have been employed for antibody detection. However
serology is not widely used.

5.Polymerse Chain Reaction (PCR):

PCR can be used for detection of HSV DNA in CSF

TREATMENT AND PROPHYLAXIS

HSV infection can be treated with acyclovir (acycloguanosine). it acts by interfering with
viral DNA synthesis by inhibiting DNA-dependent DNA polymerase. It may be used in the
form of ointment for the treatment of ocular lesions. Type 2 infection is more resistance to
treatment than type 1.Valaciclovir and famciclovir are more effective oral agents. When
resistance to these drugs develop, drug like trisodium phosphonoformate (Foscarnet) may
be useful.

Currently there is no vaccine approved available for prevention of HSV infection.

IMMUNITY: Antibody produced may not prevent recurrent infection, but can reduced
the severity of the clinical disease.

Additional Info.

Moderna’s herpes simplex virus (HSV) vaccine candidate[mRNA-1608]is an mRNA

vaccine targeted against HSV-2 disease. Moderna stated on February 18 2022, it is expected
an HSV-2 vaccine which could provide cross-protection against HSV-1 with mRNA-
1608.Moderna aims to induce a strong antibody response with neutralizing and effector
functionality combined with cell-mediated immunity.
 VARICELLA ZOSTER VIRUS

INTRODUCTION:
Varicella (chickenpox) and herpes zoster (shingles) are caused by a single virus, for which it
is named Varicella-zoster virus (VZV). Chickenpox follows primary infection in a non-
immune individual, whereas herpes zoster is a reactivation of the latent virus when
immunity falls to ineffective level. Thus, contact with either chickenpox or zoster may lead
only to chickenpox but not zoster.

MORPHOLOGY:
Varicella zoster virus is similar to the herpes simplex virus in its morphology.

CULTURAL CHARACTERISTICS:
Varicella zoster virus does not grow in experimental animals or chick embryos. The virus
was first isolated by Weller in human embryonic tissue culture. It can be grown in cultures
of human fibroblasts, human amnion or HeLa cells. The cytopathic effects are similar to, but
less marked than, those produced by the herpes simplex virus. In cultures, the virus remains
cell associated and does not appear free in the medium. By using highly specific antisera, it
is possible to distinguish between herpes virus type 1,2 & varicella zoster viruses. Only one
antigenic type of Varicella Zoster Virus is known.

VARICELLA (CHICKENPOX):
Chickenpox is one of the mildest and most common of childhood infections. The disease
may however occur at any age. Adult chickenpox which is more serious, is rather common
in some tropical areas.
The source of infection is a chickenpox or herpes zoster patient. Infectivity is
maximum during the initial stages of the disease, which the virus is present abundantly in
the upper respiratory tract. The buccal lesions which appears in the early stage of the
disease and the vesicular fluid are rich in virus content. There are no animal reservoirs of
varicella.

PATHOGENECITY:
The portal of entry of the virus is the respiratory tract or conjunctiva. After an incubation
period of about two weeks (7-23 days), the lesions begins to appear. The patient is
considered to be infectious during two days before and five days after the onset of the
lesions.
In children, there is a little prodromal illness and the disease is first noticed when the
skin lesion appears. The evolution of the rash is so rapid that the various stages – macule,
papule, vesicle, pustule and scab – cannot be readily followed in individual lesions. The rash
is centripetal in distribution and appears in crops during the first three or four days of the
disease, the rash matures very quickly, beginning to crust within 48 hours.
When Varicella occurs in adults, systemic symptoms may be severe, the rash very
profuse and the entire disease much more intense than in children. The rash may become
hemorrhagic and occasionally bullous lesion appears.
Varicella pneumonia is more common in adults and often fatal in elderly. One attack
confers lasting immunity.
Chickenpox in pregnancy: Chickenpox in pregnancy can be dangerous for both the
mother and the baby, It tends to be more severe with enhanced risk of complications like
pneumonia. The baby may develop two types of complications depending upon the period
of gestation when the woman develops chickenpox.

HERPES ZOSTER (SHINGLES, ZONA):

While varicella is a childhood disease, herpes zoster is one of old age, being common after
the age of fifty years. The disease may however, occur at any age and zoster has been
reported very rarely even in the newborn.
Herpes zoster usually occurs in persons who had chickenpox several years earlier.
The virus remains latent in the sensory ganglia. The virus is usually held in check by the
residual immunity, when the immunity has fallen into ineffectiveness levels, the virus
maybe reactivated. The reactivation is associated with inflammation of the nerve, which
accounts for the neuritic pain that often precedes the skin lesions.

PATHOGENICITY:
The lesions are identical to varicella lesions, except for their limited distribution. The rash
heals in about two weeks. Pain and paresthesia at the affected area may persist for weeks
or months. Other complications are lower motor neuron paralysis. The Ramsay Hunt
Syndrome is a rare form of zoster affecting the facial nerves.

Note: In shingles blisters appears in localized area whereas in chickenpox it covers the
entire body.
IMMUNITY:
Although the single virus is responsible for both the clinical entitles, one attack of
chickenpox confers life long immunity, but antibody fails to eliminate the virus from dorsal
root ganglia. Hence, zoster occurs in persons who have immunity to varicella.

LABORATORY DIAGNOSIS:
Diagnosis is usually clinical. Laboratory diagnosis includes:
1. Direct Microscopy: Stained smears from the base of early vesicles show
multinucleated giant cells and type A intranuclear inclusion bodies under light
microscope. Toludine blue or Giemsa stains are used for staining the smears. Herpes
particles can also be detected by electron microscopy.
2. Virus Isolation: Virus can be isolated in human fibroblast cells, human amnion, HeLa
or Vero cells. Cytopathic effect is focal with refracticle ballooned cells. However,
Varicella zoster antigen can be demonstrated in nuclear inclusions by
immunofluorescence using monoclonal antibody.
3. Polymerase Chain Reaction (PCR): DNA can be extracted from specimen and
amplified by PCR. It is useful for detection of varicella zoster virus (VZV) in CSF and
other body fluids.
4. Serology: Varicella-zoster specific IgM antibody in patient’s serum can be detected by
ELISA. Other methods used for detecting antibody are CFT, neutralization test and
immunofluorescence.

TREATMENT: Acyclovir and vidarabine are effective in the treatment of severe varicella
and zoster.

PROPHYLAXIS:
1. Varicella-zoster virus immunoglobin prepared from patients convalescing from zoster
gives passive protection in immunocompromised children exposed to infection. It is
not useful in treatment.
2. A live attenuated varicella vaccine (Oka strain) has been developed by serial passage
in tissue culture. A modified lyophilised form of vaccine is now available. It is safe and
effective. It is not considered safe in pregnancy.

ADDITIONAL INFO.
Moderna expected varicella zoster vaccine candidate mRNA-1468, designed to
express VZV glycoprotein E (gE) to reduce the rate of shingles, as stated on 18th
Feb 2022

REFERENCE: TEXTBOOK OF MICROBIOLOGY- Dr C.P Baveja 6th edition

TEXTBOOK OF MICROBIOLOGY- Ananthanarayan & Paniker’s 9th edition
TEXTBOOK OF MICROBIOLOGY & IMMUNOLOGY-Subhash Chandra parija 2nd edition.