Prof Mohamed Abdille,

BSc, MBBS,MSc(Trop Med),

LECTURE 3
MSc(Mol Bio & Gen
Engineering), PhD(Preventive
Med), FNASc, MBA
Properties of enveloped viruses
• Envelope is sensitive to
– Drying
– Heat
– Detergents
– Acid
• Consequences
– Must stay wet during transmission
– Transmission in large droplets and secretions
– Cannot survive in the gastrointestinal tract
– Do not need to kill cells in order to spread
– May require both a humoral and a cellular immune
response

Adapted from Murray, P.R. Rosenthal K.S., Pfaller, M.A. (2005) Medical Microbiology, 5 th edition, Elsevier Mosby, Philadelphia, PA
Box 6-5
 Properties of naked capsid viruses
• Capsid is resistant to
– Drying
– Heat
– Detergents
– Acids
– Proteases
• Consequences
– Can survive in the gastrointestinal tract
– Retain infectivity on drying
– Survive well on environmental surfaces
– Spread easily via fomites
– Must kill host cells for release of mature virus particles
– Humoral antibody response may be sufficient to neutralize
infection

Adapted from Murray, P.R. Rosenthal K.S., Pfaller, M.A. (2005) Medical Microbiology, 5 th edition, Elsevier Mosby, Philadelphia, PA ,
Box 6-4
 Classification of Human
Viruses

Fields Vriology (2007) 5th edition, Knipe, DM & Howley, PM, eds, Wolters Kluwer/Lippincott Williams & Wilkins, Philadelphia Table 2.1
Variations on the replication theme

From Schaechter’s Mechanisms of

Microbial Disease; 4 th ed.;
Engleberg, DiRita & Dermody;
Lippincott, Williams & Wilkins; 2007;
Fig. 31-7
 Summary: structure,
classification & replication
• Structure
– Nucleic acid in a protein shell, +/- lipid envelope
– Structure impacts on biological properties
• Classification
– Many virus families, organized by structure and
biology
• Replication
– Generic scheme
– Varying strategies depending on nucleic acid
 Summary: laboratory virology
• 4 main clinical diagnostic techniques
– Culture, serology, antigen detection, nucleic acid
detection
• Virus culture
– Cultured cell types
– Cytopathic effect
– Not all viruses can be cultured
• Virus quantitation
– Biological
– Physical
• Basic serological techniques
 Cycle of infection
Entr Sheddin
y Primary site g

Local
Lymphatic
Sprea Neuronal
d Blood (viremia)

Secondary sites
Sheddin
g
 Time course of infection; host response
infection without spread:
pro-
symptoms at
drom healing
primary site
e
infection with spread:

symptoms at
prodrome
secondary sites

host
response:
innate immunity: adaptive immunity: inflammatory;
interferon cellular, antibody immunopathogenesis

0 2 4 6 8 10 12
time (days)
 Patterns of disease

From Schaechter’s Mechanisms of Microbial Disease; 4th ed.; Engleberg, DiRita & Dermody; Lippincott, Williams & Wilkins; 2007; Fig.
31-9
 Summary:
Pathogenesis & Genetics
• Cycle of infection
• Effects on cells
– Abortive, lytic, persistent, latent, transforming
infections
• Effects on the organism
• Genetics
– Mutation, genotype, phenotype, reversion,
recombination
 For each virus, know:
• Structure (cheat sheet)
• Pathogenesis
– transmission/entry/shedding
– replication
– spread
– immune response/counter response
– damage/disease mechanism
• Diagnosis
• Treatment/prevention
– drugs
– vaccines
 Human papillomavirus
• Structure
– Small (8 kb) circular dsDNA genome, naked capsid
• Pathogenesis
– transmission by direct contact or sexual; skin, mucosa
– replication in nucleus of basal cells of epithelium; very host dependent; coupled
to epithelial differentiation
– no spread
– primarily cellular immune response
– transforming infection; warts are tumors; cervical carcinoma
• Diagnosis
– cytology (PAP smear; koilocytosis)
– immunohistochemistry
– nucleic acid
• Prevention/treatment
– recombinant subunit (VLP) vaccine
– PAP smear
– surgery
 Parvovirus
• Structure
– Small (5 kb) linear ssDNA genome, naked capsid
• Pathogenesis
– respiratory transmission
– replication in nucleus, very host dependent, needs S phase
cells or helper virus
– viremia
– antibody important in immunity
– targets erythroid lineage cells; fifth disease (symptoms
immunological); transient aplastic crisis; hydrops fetalis
• Diagnosis
– serology, viral nucleic acid
• Treatment/prevention
– none
 Polyomavirus
• Structure
– Small (5 kb) circular dsDNA genome, naked capsid
• Pathogenesis
– respiratory transmission
– replication in nucleus; very host dependent
– viremia
– persistence in kidneys; reactivation with immune compromise
– inapparent infection; hemorrhagic cystitis; PML
• Diagnosis
– viral nucleic acid
• Treatment/prevention
– cidofovir ?
 Adenovirus
• Structure
– Medium sized (36 kb) dsDNA genome, naked capsid
• Pathogenesis
– respiratory or fecal oral transmission
– replication in nucleus; moderately host dependent
– local spread; viremia
– cellular and humoral immunity important; virus encodes
countermeasures against MHC I expression and apoptosis
– direct cell damage from replication; respiratory illness,
conjunctivitis, gastroenteritis, cystitis
• Diagnosis
– culture, viral antigen detection
• Treatment/prevention
– live military vaccine
 Influenza
• Structure
– Negative sense segmented ssRNA genome, helical nucleocapsid,
enveloped
• Pathogenesis
– respiratory transmission
– replication in nucleus; budding
– no spread (usually)
– innate and antibody response important; antigenic shift and drift
– local symptoms from cell killing; systemic symptoms from immune
response; exaggerated disease in young and elderly; viral and bacterial
pneumonia complications
• Diagnosis
– culture, hemadsorbtion, viral antigen detection
• Treatment/prevention
– amantidine and rimantidine target matrix; zanamivir and oseltamivir
target NA
– killed and live vaccines need constant updating
 Paramyxoviruses
• Structure
– Negative sense ssRNA genome, helical nucleocapsid, envelope with
attachment protein and F protein
• Pathogenesis
– Transmission in respiratory droplets and fusion of virus envelope via F
protein with plasma membrane of cells in the respiratory tract
– Replication in cytoplasm, budding
– Viremia except for RSV and PIV
– Innate and antibody response important; many symptoms from immune
response: rash in measles and swelling in mumps; PIV bronchitis and
croup; RSV bronchiolitis and pneumonia in infants
– Sequelae in CNS for measles and mumps
• Diagnosis
– Serology or nucleic acid
– Measles: Koplik spots; mumps: swelling of parotid gland
• Treatment/prevention
– MMR live attenuated viral vaccine for measles and mumps, none for
RSV or PIV
 Rabies
• Structure
– Negative sense ssRNA, helical nucleocapsid, envelope
• Pathogenesis
– Transmitted by bite of rabid animal
– Replication in cytoplasm; budding
– Spread by axonal transport to brain; long incubation period
– Fever, nausea, hydrophobia, coma
– Almost always fatal
• Diagnosis
– Viral antigen or nucleic acid, Negri bodies
• Treatment/prevention
– Inactivated viral vaccine for humans after exposure, live
virus vaccine for animals
 Rotavirus
• Structure
– Naked double shell capsid
– 11 segment double stranded RNA genome
• Pathogenesis
– Fecal oral transmission
– Replication in cytoplasm
– Fever, vomiting, diarrhea in infants and young children
– Incubation period less than 48 hr, highly infectious
– Infection of intestinal epithelium causes loss of electrolytes and
prevents readsorption of water
– Long term immunity; asymptomatic infection in adults
• Diagnosis
– viral antigen detection
• Treatment/prevention
– RotaTeq live, oral vaccine
Summary
 Paramyxoviruses
Structure
• Negative sense ssRNA genome, helical nucleocapsid, envelope
with attachment protein and F protein
Pathogenesis
• Transmission in respiratory droplets and fusion of virus envelope
via F protein with plasma membrane of cells in the respiratory tract
• Replication in cytoplasm, budding
• Viremia except for RSV and PIV
• innate and antibody response important; many symptoms from
immune response: rash in measles and swelling in mumps; PIV
bronchitis and croup; RSV bronchiolitis and pneumonia in infants
• Sequelae in CNS for measles and mumps
Diagnosis – serology or nucleic acid
• Measles Koplik spots; mumps swelling of parotid gland
Treatment/prevention
• MMR live attenuated viral vaccine for measles and mumps, none
for RSV or PIV
 Rabies
Structure
• Negative sense ssRNA, helical nucleocapsid, envelope

Pathogenesis
• Transmitted by bite of rabid animal
• replication in cytoplasm; budding
• Spread by axonal transport to brain; long incubation period
• Fever, nausea, hydrophobia, coma
• Almost always fatal

Diagnosis
• Viral antigen or nucleic acid, Negri bodies

Treatment/prevention
• Inactivated viral vaccine for humans after exposure, live virus vaccine for
animals
Time course of Rabies infections
 Rotavirus
Structure
• Double stranded, 11 segment RNA genome, two protein coats

Pathogenesis
• Fecal oral transmission
• replication in cytoplasm
• Epidemic diarrhea in infants and young children- fever,
vomiting, diarrhea
• Incubation period less than 48 hr, highly infectious
• Infection of intestinal epithelium causes loss of electrolytes
and prevents re-adsorption of water

Diagnosis
• culture, viral antigen detection

Treatment/prevention
• RotaTeq live, oral vaccine
 Enteroviruses
Structure
• Positive sense ssRNA genome, protein coat

Pathogenesis
• Fecal oral transmission
• replication in cytoplasm
• Viremia to diverse target tissues. Viruses very cytopathic, killing cells
they infect
• Infections often asymptomatic; polio causes paralytic poliomyelitis;
rhinoviruses restricted to upper respiratory tract, common cold;
caliciviruses diarrhea

Diagnosis
• Serology and nucleic acid

Treatment/prevention
• Only polio vaccines, Salk and Sabin
 Arboviruses
Structure
• Positive sense ssRNA genome, icosahedral nucleocapsid, enveloped

Pathogenesis
• Transmitted by bite of insect from host species; sylvan and urban
cycles
• replication in cytoplasm; budding
• Viremia to target tissue
• Influenza-like initial symptoms; different viruses cause encephalitis,
hemorrhagic fever, hepatitis, rash, arthritis

Diagnosis
• Serology and nucleic acid

Treatment/prevention
• No human vaccines, except yellow fever virus live attenuated vaccine,
control of insect population
Arboviruses: Vectors, Hosts, and Diseases
 Rubella virus
Structure
• Positive sense ssRNA genome, helical nucleocapsid, enveloped

Pathogenesis
• respiratory transmission
• replication in cytoplasm; budding
• Viremia
• Mild rash in adults; congenital rubella syndrome (CRS) after
infection in first trimester when virus passes the placenta and
infects fetus
• CRS- deafness, blindness, mental retardation

Diagnosis
• Nucleic acid, viral antigen detection

Treatment/prevention
• MMR live attenuated virus vaccine
 RNA Hepatitis Viruses
Structure
• Various different families for HAV, HCV, HEV, HGV all positive sense ssRNA

Pathogenesis
• HAV and HEV fecal oral then viremia; others sexual and blood borne, viremia
• Liver is target organ; most infections are subclinical, acute infections differ in
onset and severity.
• HAV and HEV cause hepatitis with no carrier state; others cause hepatitis
with chronic infection and possible carcinoma
Liver damage due to cell mediated immune response

Diagnosis
• viral antigen detection, nucleic acid

Treatment/prevention
• HAV killed virus vaccine; HCV alpha-interferon effective for some serotypes
 HG
V

///////////hepe
Flav
i
 Hepatitis B Virus
• Structure
– Small (3.2 kb) circular partially dsDNA genome, envel. capsid
• Pathogenesis
– Sexual, parenteral, and perinatal transmission
– Replication via an RNA intermediate (reverse transcriptase)
– Tropism for liver
– Acute vs. chronic infections occur: highly age dependent
– Chronic infections are a major cause of PHC
• Diagnosis
– Multiple serological components (viral proteins and anti-bodies)
• Treatment/prevention
– Subunit vaccine (based on HBsAg)
 Hepatitis B virus

CDC website: http://www.cdc.gov/ncidod/diseases/hepatitis/slideset/hep_b/slide_1.htm

 Immunological events of acute vs. chronic
HBV infection

A) Acute B) Chronic

From Murray et. al., Medical Microbiology 5 th edition, 2005, Chapter 66, published by Mosby Philadelphia,,
 Clinical interpretation of the Hepatitis B
antigen panel

CDC WEB site: http://www.cdc.gov/ncidod/diseases/hepatitis/b/Bserology.htm

 Hepatitis D Virus
• Structure
– Very Small (1.7 kb) circular ssRNA genome, envel. capsid
• Pathogenesis
– Sexual, parenteral, and perinatal transmission
– Replication by RNA-directed RNA Pol (Host RNA Pol II)
– Requires concurrent HBV infection (needs it for HBsAg)
– HDV greatly exacerbates liver damage caused by HDV
– Chronic infections are a major cause of PHC
• Diagnosis
– Serologically for HDV delta antigen
• Treatment/prevention
– Subunit vaccine for HBV prevents productive infection
 HIV
• Structure
– 9.0 kb diploid ssRNA genome, envel. capsid
• Pathogenesis
– Sexual, parenteral, and perinatal transmission
– Replication by cDNA intermediate (reverse transcriptase)
– Replication cycle requires the DNA intermediate to integrate
– HIV establishes a persistent infection that ultimately reduces CD4
helper T cell population
– During course of infection, tropism shift from M-tropic to T-tropic
• Diagnosis
– Serologically for antibodies against HIV antigens (gp120)
• Treatment/prevention
– Antivirals (HAART)
 HIV-1 Replication Cycle
Reverse
Integratio Assembl
Transcription
n y
Integras
Attachmen Reverse e
t Transcriptase

CD Uncoatin Buddin
4 g g

CCR
5

Maturation
Proteas
e
CXCR
4 Beth D. Jamieson, Ph.
Natural Course of HIV-1 Infection
 Herpesviruses
Herpes simplex I & II (cold sores, genital herpes)
Varicella zoster (chicken pox, shingles)
Cytomegalovirus (microcephaly, infectious mono)
Epstein-Barr virus (mononucleosis, Burkitt’s lymphoma)
Human herpesvirus 6 & 7 (Roseola)
Human herpesvirus 8 (Kaposi’s sarcoma)
 Human Herpesviruses
Virus Subfamily Disease Site of Latency

Herpes Simplex Virus I α Orofacial lesions Sensory Nerve Ganglia

Herpes Simplex Virus II α Genital lesions Sensory Nerve Ganglia
Varicella Zoster Virus α Chicken Pox Sensory Nerve Ganglia
Recurs as Shingles

Cytomegalovirus β Microcephaly/Mono Lymphocytes

Human Herpesvirus 6 β Roseola Infantum CD4 T cells
Human Herpesvirus 7 β Roseola Infantum CD4T cells

Epstein-Barr Virus γ Infectious Mono B lymphocytes, salivary

Human Herpesvirus 8 γ Kaposi’s Sarcoma Kaposi’s Sarcoma Tissue
A study of HSV-2 recurrence in
women
Vaccines – live or killed?

From Murray et. al., Medical

Microbiology 4 th edition, 2004,
Chapter 15, published by Mosby
Philadelphia,,
 FDA approved antiviral drugs
Table 50-2

* Also active against varicella-zoster virus T

Topical use only From Murray et. al., Medical Microbiology 5 th edition, 2005,
Chapter 50, published by Mosby Philadelphia,,
 Non-equilibrium human virus

1. Virus has a stable relationship with an animal

host.

2. Virus can be strikingly lethal since it hasn’t

evolved to coexist with humans.

3. Virus will be in genetic flux until it reaches

equilibrium or human infection chain is broken.

4. HIV, Ebola virus, Hantavirus and Influenza virus

examples