Chapter 18- Animal Viruses

A. Cultivating Animal Viruses

• Must be done inside living tissues or cells!

• Embryonated eggs

• Must be kept otherwise sterile

• Monitor by “candling” eggs to view health of embryo

• Tissue culture cells

• Grow cells from animal source in flasks

• Many form monolayers

• Cell lines are “immortalized” and can replicate indefinitely

o Primary cell cultures can only undergo about 50 cell divisions before
senescence

B. Penetration by Animal Viruses

• Direct penetration by nucleic acid by naked viruses

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• Membrane fusion- enveloped virus fuses with plasma membrane & nucleocapsid
enters host

• Endocytosis

o Naked viruses also enter their hosts this way

o In enveloped viruses, enzymes are present to degrade the lipid bulyers of botht he
vesicle and the envelope to release the nucleocapsid

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C. Consequences of Animal Virus Infections

• Cytocidal Infections- host cell death

o May be preceded by cytopathic effects (CPE)
• Inclusion bodies
• Syncitium formation (giant fused cells)
• Cell detachment and rounding

• Persistent infections
o Slow release of virus so host cell is not lysed
§ HIV, serum hepatitis (Hepatitis B)
o Latent infections
§ Virus is dormant and no symptoms, virus or antibody detectable
§ Herpes simplex (cold sores; genital herpes)
§ Varicella-zoster (chicken pox / shingles)
§ Mechanisms varied and poorly understood

• Viruses and Cancer

o Transformation refers to uncontrolled growth by host cell = cancer

o Neoplasia refers to abnormal new cell growth

o Cell growth controlled by at least 2 types of genes:

§ Proto-oncogenes promote growth

§ Tumor suppressor genes suppress growth

§ Changes in one or both lead to transformation

o Metastasis refers to spread of transformed cells

o 1. Epstein-Barr Virus
§ Infects B lymphocytes
§ Mononucleosis
§ Burkitt’s Lymp homa -central & west Africa
• Malignant tumor of jaw and abdomen
§ Nasopharyngeal carcinoma in Southeast Asia

o 2. Papillomavirus and cervical cancer

o 3. Hepatitis B virus and some liver cancer

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D. Human Immunodeficiency Virus

• AIDS (acquired immunodeficiency syndrome) first described in 1981

• HIV classified as a lentivirus

• The disease originated in central Africa perhaps in 1950’s

o Simian Immunodeficiency Virus similar to HIV, hence HIV may have

originated in African primates

o Spread from Africa to the Carribean, and then to the US

o Acquired by direct exposure to body fluids

o May be acquired through breastmilk

• Spike protein gp120 binds CD4 receptor molecule on CD4+ lymphocytes,

macrophages, & dendritic cells

• Internalization and uncoating of the virus within host cells

• Reverse transcriptase makes DNA copy of genome

• Viral DNA inserts into host cell genome

• Activation of provirus results in rapid production of virus (107 particles /day)

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E. HIV Pathogenesis

• Mechanism poorly understood

• Both apoptosis & direct CPE may contribute

• Results in depletion of CD4+ lymphocytes

• Generalized dysfunction of uninfected cells also evident

• Drop from 1000 to 100 CD4+ lymphocytes / mm3 of blood

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F. HIV & Immunodeficiency

• CD4+ T cells are fundamentally important in immune respons e

• Depletion of these cells makes individual susceptible to variety of microbial agents

• Microbes usually harmless can cause life-threatening illness

G. Progression of ( HIV) Disease

• 1. Acute HIV syndrome initially

o 2-6 weeks after exposure; Virus replicates and is abundant in blood
o Fever, sore throat, pharyngitis, rashes, lymphadenopathy

• 2. Clinically latent stage

o May last 10 years or longer
o Virus particles disappear from body fluids
o AIDS-Related Complex (ARC) in some

• 3. Clinical AIDS
o Includes weight loss (cachexia) & diarrhea

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H. Relative Dose: The Swimming Pool Analogy

• Each virus has a different size relative dose (or amount of virus present to cause a
likely infection)

• Hepatitis B Virus:

1 ml infected blood added to full swimming pool of water, and 1 ml of that were injected
into an individual, they WOULD become infected

• HIV:

1 ml of blood from AIDS patient added to quart of water, and 1 ml of that were injected
in individual, they have a 1 in 10 chance of becoming infected

I. Herpes simplex virus

• Virus replicates in epithelial cells of skin

• Some virus particles move to sensory nerve endings in that part of skin

• Virus moves down nerve cell to trigeminal ganglia of the nerves and persists there as
circular DNA in dormant phase

• Stimulus causes virus to move back down neuron to epithelium and replicate
o Stress, UV light, fatigue, other infections

• HSV-1

• dsDNA virus with icosahedral capsid & envelope

• “Herpes” derived from Greek word meaning “to creep”

• Transmission through direct contact

• Blisters appear in epidermis and surface mucous membrane and usually heal
within a week

• Latent state lasts a lifetime

• Infections involving the eye can cause blindness

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J. Influenza

• Caused by Orthomyxovirus (not by Haemophilus influenzae)

• Enveloped ssRNA genome in 8 segments

o Negative-stranded genome

• Reservoir in rural China

• Chickens, pigs and humans live in close proximity in crowded conditions

o Jumps from birds to pigs to humans

§ Swine flu, avian flu

o Zoonosis

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• Influenza Hemagglutinin (HA)

• Binds to receptor on erythrocytes, causing them to clump

• Receptor-binding protein

• Antibodies to HA block attachment, neutralizing virus

o Immunity to flu due to antibodies to HA

• Synthesized as single polypeptide

o Cleaved by host protease to form HA1 & HA2 polypeptides
o Essential for pathogenicity

• Neuraminidase (NA)

• Removes terminal sialic acids from host glycoproteins

• Seems important in virus assembly and release

• May allow budding of virus from host cell without attaching to host cell sialic
acids which might prevent its escape

K. Antigenic Drift and Influenza

• Mutations in HA and NA genes occur at higher frequency than normal

• Missense mutations result in amino acid substitutions

• Some mutations make the proteins no longer recognizable by the immune response

• Thus, virus changes regularly, making it necessary to be vaccinated against influenza

each year, as you are likely to encounter a flu virus different from those in previous
years

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L. Antigenic Shift and Influenza

• Otherwise known as “why we have to get flu shots every year”

• Major change in the HA or NA of a flu virus

• Results from co- infection of same cell with 2 different flu strains

• Because genomes are segmented, the segments can assort independently

• May yield new strain with HA from one strain and NA from the other strain

M. Pathogenesis of Influenza

• Transmitted by inhaling droplets released by coughing & sneezing

• Infects mucosal epithelium of upper respiratory tract

• May invade lungs

• Attaches via HA
• HA Variability affects host specificity

• Entry by endocytosis
• pH drops & virus enters cytoplasm

• Replication in nucleus

• Assembly and budding

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N. Common Cold or the Flu?

Symptom Cold Flu

Fever rare common

Headache rare common

Malaise mild often extreme

Nasal discharge abundant usually not abundant

Sore throat common much less common

Vomiting/diarrhea rare common

O. Complications associated with flu & other viral diseases

• Reyes Syndrome
• Persistent vomiting, convulsions, delirium & coma
• Brain & liver damage
• Linked to taking aspirin or related products during infection

• Guillain-Barré Syndrome
• French polio; follows vaccine or infection
• Weakness in extremities & sensory loss
• Damage to Schwann cells & demyelination
• Not permanent

P. Treatment for flu and viral diseases

• Amantadine:
o Active after adsorption but before transcription
o Useful for prophylaxis or within 1st 48 hr
o Blocks entry of virus into cytoplasm

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Q. Prion Diseases

• Bovine Spongiform Encephalopathy

o Also called Mad Cow Disease
o Beef exports excluded from Europe where outbreaks of disease are epidemic
o Also blood donations from people traveling in outbreak areas are not accepted

• Scrapie in sheep (was described in 1700’s)

• Kuru
o New Guinea
o Cannibalism of the dead of tribe

• Creutzfeld-Jakob Disease (CJD)

o Agent withstands treatments that inactivate most other diseases
o Can be inherited
o Can be acquired
o Transmission best between same species

• Infectious agent: what is a prion?

o Passes through filters that retain viruses
o Withstands treatments that inactivate most viruses, including very high
temperatures
o Insensitive to nucleases
o Made entirely of protein

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Prion pathogenesis

• Most cells contain a gene encoding a protein that is very similar to the prion protein
o Normal protein vs “rogue” protein
o Normal isoform = PrPC
o Scrapie isoform = PrPSc

• Protein found on most cells in mammals but function is not known

• Inherited cases of CJD occur with naturally occurring mutation in gene for this
protein

• Since the prion originates from the host gene, only distinction between Scrapie and
CJD is based on the host gene

• Prions non- immunogenic

• PrPC seems to help protect brain from dementia and other degenerative problems
associated with old age

• Mouse mutants lacking PrPC gene develop dementia more rapidly

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