Microbiology (Lecture)

PARVOVIRUS, PAPILLOMAVIRUS, POLYOMAVIRUS

Dr. Liela D. Ferrer | April 23, 2021 | Topic 12
Table 1. Diseases caused by Parvovirus (B19)
Overview Syndrome Host or Clinical features
condition
th
I. General Characteristics Erythema Children (5 Cutaneous rash
II. Parvovirus infectiosum disease) Arthralgia-arthritis
III. Papillomavirus Transient Underlying Severe acute
IV. Polyomavirus anaplastic crisis hemolysis anemia
Purple red cell Immunodeficiency Chronic anemia
anaplasia
CHARACTERISTICS Hydrops fetalis Fetus Fatal anemia
Ø Viruses having a DNA genome
Ø All 3 have icosahedral capsid, circular DNA TH
ERYTHEMA INFECTIOSUM (5 DISEASE)
Ø All DNA viruses have dsDNA EXCEPT Parvoviruses Ø Children of early school age; occasionally adults
(ssDNA) Ø Incubation period: 1-2 weeks
Ø All replicate in the nucleus Ø Viremia 1 week after infection à persist for 5 days à
Ø All are non-enveloped or naked virus present in nasal washes and gargle specimens
• Pharynx: most probable site of viral shedding
PARVOVIRUS Ø Symptomatic
Ø Smallest and simplest of all virus Ø Biphasic à lytic stage (contagious stage) and
Ø Single-stranded DNA; no virion polymerase; immunologic
icosahedral; naked Ø Generalized erythematous rash is most prominent in
Ø VP2: major capsid protein the face à “slapped cheek” appearance; lace-like
Ø NSP (NS-1): required for replication rash on limbs or trunk
Ø Highly resistant to inactivation Ø Arthralgia (hands and knees): adults
Ø Replicates only when a cell is in S phase (DNA Ø Treatment
synthesis) • Transient anaplastic crisis: symptomatic; may
Ø Defective virus à requires a helper virus for require blood transfusion
replication (Herpesvirus or Adenovirus) • Immunoglobulin preparations containing
Ø Human pathogen-B19: with tropism for RBC neutralizing antibodies for immunocompromised
progenitors and those with anemia
CLASSIFICATION OTHER CLINICAL SYNDROMES
1) Parvovirinae 1) Arthritis in adults: resemble rheumatoid arthritis
a) Genus Erythrovirus 2) Pure red cell aplasia: persistent infections and
ü Parvovirus B19 (type 1) chronic anemia in immunocompromised patients
ü Strain K71 (type 2): recently identified 3) Hydrops fetalis
ü Strain V9 (type 3): human genotype nd
• Result of severe fetal anemia especially in 2
b) Genus Bocavirus: newly discovered
trimester
c) Genus Dependovirus
• Congenital heart failure
ü AAV-2 à defective virus à depend on a th
helper virus (Adenovirus or Herpesvirus) for • Fetal death before 20 week of pregnancy
replication • No congenital abnormalities
2) Densovirinae: infect insects
HUMAN BOCAVIRUS RESPIRATORY INFECTION
B19 VIRUS Ø Prevalent in children with acute wheezing
Ø Only member that can cause infections in humans Ø Often found in mixed infections with other viruses
Ø Cellular receptor: blood group P antigen (mature Ø Pathogenesis is unknown
erythrocytes, endothelial cells, placenta, Ø Mode of transmission: probable respiratory route;
megakaryocytes, fetal liver, heart) also detected in stool and serum samples
Ø Has tropism for
1) Erythroblasts in the basement membrane à LABORATORY DIAGNOSIS
aplastic anemia Ø Most sensitive tests detect viral DNA à PCR (most
2) Endothelial cells in blood vessels à rash sensitive), in situ hybridization of fixed tissue
Ø Mode of transmission Ø Serology
1) Respiratory route: primary • B19 IgM antibody: recent infection; present 2-3
2) Blood transfusion months after infection
3) Vertical/transplacental transmission
Ø No evidence of virus excretion in feces or urine PREVENTION
Ø Humans: natural reservoir Ø No vaccine available
Ø Principal targets: immature cells in erythroid lineage Ø Good hygiene
Ø Replication à death of infected cells à cessation of Ø Standard infection control practices for health workers
RBC production

Irelia: The Blade Dancer 1

 Microbiology (Lecture)
PARVOVIRUS, PAPILLOMAVIRUS, POLYOMAVIRUS
Dr. Liela D. Ferrer | April 23, 2021 | Topic 12
PAPILLOMAVIRUS 3) Sexual contact
Ø Virion: naked, icosahedral 52-55 nm in diameter; 4) Vertical transmission
contain covalently linked circular DNA
Ø Small, icosahedral capsid, nonenveloped, dsDNA CLINICAL INFECTIONS
genomes 1) Skin warts: serotypes 1-4
Ø Capsid: contains 72 capsomeres in skew 2) Laryngeal papillomas: serotypes 6 and 11
arrangement; each capsomere is composed of 2 viral 3) Anogenital warts (condyloma acuminate): 6 and 11
proteins 4) Cervical, oropharyngeal and penile carcinomas:
• L1: major protein 16 and 18
• L2: major capsid protein 5) Epidermodysplasia verruciformis
Ø Composition: DNA = 10%, proteins = 90%
Ø Genome: non-segmented, circular or supercoiled LABORATORY DIAGNOSIS
dsDNA 1) Cytology
Ø Proteins: 2 structural proteins, cellular histones 2) DNA molecular probe
Ø Causes lytic, chronic, latent and transforming 3) PCR
infections depending host cell 4) Southern blot
1) Early genes (E1-E7)
• Facilitate replication of viral genome TREATMENT
• Codes for nonstructural protein 1) Removal of the lesion
2) Late genes (L1 and L2) • Surgical excision
• Viral attachment protein • Application of caustic agents
• Codes for structural protein • Cryosurgery
• Electrocautery
HUMAN PAPILLOMAVIRUS • Laser therapy
Ø Former members of Papovaviridae 2) Anti-HPV drugs or immunomodulating drugs
Ø Double-stranded DNA, icosahedral, naked • Imiquimod
Ø Oncogenic à E6 and E7 proteins • Systemic or intralesional alpha interferon
Ø Do not grow in cell culture • Topical idoxuridine
Ø Capsomere form regular pointed star-shaped head • Cidofovir
Ø Tissue tropism à epithelial cells of skin and mucous
membranes PREVENTION
Ø Encodes 7 early genes (E1-E7) and 2 late structural Ø Quadrivalent and bivalent HPV vaccine
genes (L1 and L2) à bind with growth suppressor • Contain virus-like particles composed of HPV L1
proteins proteins
• E6 protein binds to p53 • Quadrivalent: HPV 6, 11, 16, and 18
• E7 protein binds to p105RB • Bivalent: 16 and 18
Ø In permissive cells à cause lytic infections
Ø In non-permissive cells à abortive, latent, persistent, POLYOMA VIRUSES
immortalizing infections Ø Formerly part of Papovaviridae (no longer exists) à
Ø Important characteristics share many similarities with Papillomaviridae due to
• Stimulate cell DNA synthesis its transforming capabilities
• Significant cause of human cancer Ø Circular, dsDNA genome; icosahedral; naked
• Viral oncoproteins interact with cellular tumor Ø Produced malignant transformation of infected cells
suppressor genes Ø Virion: icosahedral, 40-55 nm in diameter
Ø Classification Ø Composition: DNA = 10%; proteins = 90%
• Alpha papillomavirus Ø Genome: covalently link circular, dsDNA
• Beta papillomavirus Ø Proteins: 3 structural proteins, cellular histones,
• Gamma papillomavirus condense DNA in virion
• Mupapallomavirus Ø Envelope: none
• Napapapillomavirus Ø Replication: nucleus

PATHOGENESIS VIRAL PROTEINS-TUMOR ANTIGENS

Ø Infect and replicate in squamous epithelium (warts) or 1) LT (large tumor antigen)
mucous membranes (papillomas) ü Key role promotes DNA synthesis: needs to
Ø Recurrence is common be in S-phase for this to begin
Ø Innate immunity and cell-mediated immunity in ü Modulates and stimulate progression of cell
resolution and control of infection cycle
ü Induces
MODES OF TRANSMISSION § DNA replication
1) Direct contact § Expression of the late MRNA that
2) Minor skin abrasions encodes the viral capsid proteins:

Irelia: The Blade Dancer 2

 Microbiology (Lecture)
PARVOVIRUS, PAPILLOMAVIRUS, POLYOMAVIRUS
Dr. Liela D. Ferrer | April 23, 2021 | Topic 12
expression of the late genes that
accumulate of viral capsid proteins in
cell cytoplasm and capsid components
enter the nucleus encapsidate new viral
genomic DNA
ü Achieved by a 2 prong attack
1) Inhibiting tumor suppressing genes p53
and retinoblastoma (pRB) family
2) Stimulating cell growth pathways by
binding cellular DNA: ATPase-
helicase, DNA polymerase alpha
association, binding of transcription
preinitiation complex factors
*Note. Abnormal stimulation of the cell cycle: powerful force
of oncogenic transformation
2) ST (Small tumor antigen)
ü Activates several cellular pathways that stimulate
cell proliferation
ü Commonly target protein phosphatase 2A
(PP2A): a key multisubunit regulator pathways
including Akt, mitogen-activated protein kinase
(MAPK) pathway, stress-activated kinase (SAPK)
pathway
MECHANISMS OF VIRAL RELEASE
Ø Some express proteins facilitate cell exit such as the
agnoprotein or VP4
Ø In some cases, high levels of encapsidated virus
result in cell lysis
REPLICATION CYCLE
Ø Entry into a host cell
Ø Cellular receptor: sialic acid residues of glycans
(gangliosides)
Ø VPI: mediates attachment to host cells by binding to
sialylated glycans on the cell surface
• JC virus: require interaction with the 5HT2A
receptor
• Merkel cell virus: with heparin sulfate
Ø After binding, endocytosed and enters the ER (a
behavior unique among known non-enveloped virus)
• Virion particle: released from ER into the cell
cytoplasm
• Genome released from capsid: disrupted by
isomerase enzymes
CAPSID PROTEINS
Ø Expressed from the late region of the genome
Ø Polyomavirus capsid consists of
1) VP1: 1 major capsid protein
• Pentamers form the closed icosahedral viral
capsid
• Responsible for receptor binding
• Surface contains a sialic binding site
2) VP2 and VP3: minor capsid proteins
• Capsid proteins facilitate entry and uncoating
• Merkel cell polyomavirus: do not encode or
express VP
AGNOPROTEINS
Ø A small multifunctional phosphor-protein
Irelia: The Blade Dancer
Ø Location: late coding part of the genome of some
polyomaviruses
Ø Most notably BV viruses, JC viruses, and SV40
Ø Function: essential for proliferation in the viruses;
involved in regulating the viral life cycle particularly
a) Replication
b) Viral exit from the host cell
GENOME
Ø 2 regions
1) Early region: necessary for replication of viral
DNA in permissive cells
2) Late region: synthesis of coat protein
Ø Stimulate viral oncoproteins interact with cellular
tumor suppressor proteins
Ø Important model tumor virus
Ø May cause human cancer
TYPES
1) BK virus: acute hemorrhagic cystitis in bone marrow
transplant patients and ureteral stenosis
2) JC virus: progressive multifocal leukoencephalopathy
à destruction of oligodendrocytes; possible role in
colorectal cancer
3) Simian 40 virus: animal pathogen
BK AND JC VIRUSES
Ø Usually early childhood
Ø May persist in kidney, lymphoid tissues of healthy
individuals
Ø Associated with human brain tumors
KI AND WU VIRUSES
Ø Discovered in 2007 in nasopharyngeal aspirates from
children with respiratory infections
Ø Widespread
Ø Occur likely in childhood
MERKEL CELL POLYOMAVIRUS (2008)
Ø Merkel cell carcinoma: skin tumor of neuroendocrine
origin
SV40 VIRUS
Ø Fecal-oral route
Ø DNA detected in brain tumors, mesotheliomas, bone
tumors, and lymphomas
REFERENCES
th
Ø Jawetz’ Medical Microbiology 27 Edition
Ø Dr. Ferrer’s presentation
Ø Beshywap transcript
3
 MICROBIOLOGY
Orthomyxovirus
Dr. Quiles
ORTHOMYXOVIRUS NEURAMINIDASE
 ssRNA, segmented (8 segments), helical, enveloped  Cleaves sialic acid  facilitates spread of infection
 Negative sense o When sialic acid is cleaved, there will be more HA that will attach
o They cannot undergo transcription unless they produce to the sialic acid, therefore it will facilitate the spread of infection
to the adjacent cells
their own RNA-dependent RNA polymerase because they
cannot access the host cell polymerase  Degrades protective layer of mucus in the respiratory tract
 3 Groups: Influenza A, B, & C  distinguished thru the internal  Antibodies produced are non-neutralizing but reduce the spread of
ribonucleoproteins  nucleocapsid & matrix proteins infection
o Influenza C is the most antigenically stable group of
orthomyxovirus ANTIGENIC VARIATION
1. Antigenic Drift
o Minor change
SEGMENTS
o Result of changes in the amino acid sequence in the H
 Segments 1, 2, 3 – polymerase component
antigen  point mutation
 Segment 4 – HA  Hemagglutinin
Take note that N antigen can also go into antigenic
 Segment 5 – NP  Nucleocapsid
drift but only in Influenza A
 Segment 6 – NA  Neuraminidase
 Segment 7 – M1  matrix protein o Occur for both Influenza A & B
M2  membrane protein o Outbreaks & Epidemics
M1 – structural protein and promotes assembly of the virus
M2 – facilitates uncoating and HA production
2. Antigenic Shift
– target of amantadine and rimantadine
o Result of the genetic reassortment involving the H antigen
or N antigen (high frequency recombination)
 Segment 8 – NS1 & NS2  non-structural proteins
o Occur only in Influenza A
o Responsible for Pandemics & Epidemics
HEMAGGLUTININ (HA)
o Cyclic  occurs every 7 to 10 years
 Binds with the receptors (Sialic acid and Neuraminic acid)
o Both receptors are acidic which means that if these
Picture on Left:
receptors are alkaline, the HA will not bind to it
Reservoir of Influenza A
 Promotes fusion to host cell membrane
 Target of neutralizing antibodies Influenza virus have
 Agglutinates red blood cells  basis for hemagglutination inhibition antigenic variations
test because they have sources
that are different from
human (which are
animals). Any of the virus
within an animal reservoir
that would combine and
undergo genetic
reassortment with humans
will create a “new
serotype”

Picture on Left:
Antigenic Shift

The avian strain will

combine with the human
Picture Above: Illustration of Influenza Virus strain to create a new
Hemagglutinin is colored in purple. Neuraminidase is colored in yellow human strain which
Inside the virus are the 8 segments. Between the capsid and the envelope is creates a new serotype
the matrix protein

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“Strength In Knowledge” BESHYWAP 1
 MICROBIOLOGY
Orthomyxovirus
Dr. Quiles
High Pathogenicity Avian Influenza:
 1st recognized in Italy in 1878
 Extremely contagious in birds
 Rapidly fatal
 Mortality: Almost 100 % in just a matter of days
 H5N1 Influenza in 1997 in China

NOVEL INFLUENZA A H1N1 (2009)

 A unique combination of influenza virus genes never previously
identified in either animals or people
 Previously referred to the virus as a swine origin influenza virus
 Quadruple virus: 2 avian, 1 human, 1 swine virus genes
 Resistant to amantadine & rimantadine
Picture Above: Origin of Human Pandemic Influenza A viruses  Susceptible to oseltamivir & zanamivir
Verbatim recordings: Initially it started with H2N2, involving the H and the N
antigen. In 1900’s H3 at tsaka N8, hindi na kapareho ng H2N2. So yung sa H
at tsaka N nagkapareho nagkaroon ng change. Tapos nung 1918, H1N1 also
known as “Spanish Flu”, maraming namatay diyan. Nag evolve yan through
the years. Nung 1977, H1N1 or the “”Russian Avian Flu.”
H2N2 nung 1957 na kapareho ng 1889. Take not children, in the future ano
pa ang pwedeng lumabas na mga serotypes? Eto pa rin ang the one in
question

NOTE:
o Among the Influenza viruses, only A and B undergoes antigenic
variations
o Influenza A can undergo both antigenic drift & antigenic shift
o Influenza B only undergoes antigenic drift

INFLUENZA A Picture Above: Genetic Evolution of H7N9 Virus in China, 2013

 Subtypes are based on antigenic variation of the surface Verbatim recordings: From the ducks, wild birds & domestic poultry,
glycoproteins nagsama-sama lahat yan, ano ang naging resulta? This is now a new strain
 It involves the hemagglutinin and neuraminidase that may infect humans
 Current subtypes of Influenza A in humans are H1N1 & H3N2
 Influenza A H1N1 & H3N2 & Influenza B strains are included each INFLUENZA
year in the vaccine  Commonly called “the flu”
 Highly contagious
INFLUENZA VIRUS CLASSIFICATION  Illness ranges from mild to severe with life-threatening complications
Influenza A Classification:
 Type A Flu Symptoms:
 Place of original isolationDate of original isolation  Fever and/or chills
 Antigen (HA or NA)  Headache
 E.g. A/Bangkok/ 1/79 (H3N2)  Nasal congestion
 Sore throat & dry cough
Influenza B Classification:  Myalgia or body aches
 Type B  Malaise or tiredness
 Geography  GI symptoms – intestinal flu
 Date of isolation
 E.g. B/Singapore/ 3/64
Question: Why is there no H or N antigen in classification of
Influenza B? Since it only undergoes antigenic drift, it only
involves H antigen, meaning to say, it is automatically understood Picture on Left:
that the antigen involved is the H antigen 1918 Spanish Flu
(Pandemic) H1N1
AVIAN FLU
 Caused by 15 subtypes of Influenza A virus subtypes of avian 20-30 million people may
influenza have died worldwide
 Can be of low pathogenicity or high pathogenicity

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“Strength In Knowledge” BESHYWAP 2
 MICROBIOLOGY
Orthomyxovirus
Dr. Quiles
Treatment:
 Amantadine – inhibit uncoating
– target  M2 proteins
Picture on Left:
– Influenza A
Mode of
 Rimantadine – alternative drug (same MOA as Amantadine)
Transmission of
 Oseltamivir – inhibit N antigen  H antigen binds to sialic acid 
Influenza
clumps  no release of virus
– Influenza A & B
– Effective during the 1st 24-48 hours of infection
 Zanamivir – same as Oseltamivir
– inhalation
 Influenza virus can easily get inside the body through the ICAM1
REFERENCES
receptors which is readily found in our noses
 Lecture notes
Complications:  Microbiology Manual (2019)
 Primary Influenza Pneumonia  Dr. Quiles Recordings
 Secondary Bacterial Pneumonia
Among the different bacteria that can cause pneumonia, the most common “God pours His love into our hearts to flow out to others’ lives”
bacteria that is associated with influenza are Staphylococcus aureus,
Haemophilus influenzae, & Pneumococcus. But among the 3, the most
common is Staphylococcus aureus

 Gullain-Barre syndrome
 Reye’s syndrome
 Myositis
 Myocarditis
 Encephalitis
 Pericarditis

Most common cause of death in influenza virus infection is Pneumonia

REYE’S SYNDROME
 Encephalopathy & liver degeneration
 Common complication in children particularly Influenza B, A & VZV
o Among Influenza A and B, it is Influenza B that is commonly
associated with Reye’s syndrome
 Associated with intake of aspirin

INFLUENZA VACCINE
 Killed vaccine
 Contains 2 strains of Influenza A & only 1 strain of Influenza B
o New vaccines today are a combination of 2 strains of Inf.
A & 2 strains of Inf. B
 Reformulated each year to contain the current antigenic strain
 Protection lasts only for 6 months
o But according to Jawetz: 1 year

PREVENTION & TREATMENT

Prevention: Vaccine
 Trivalent or Quadrivalent inactivated or split type vaccine
o Trivalent – 2 A strains; 1 B strain
o Quadrivalent – 2 A strains; 2 B strains
o Take note this is killed or inactivated vaccine

 Live attenuated influenza vaccine (LAIV)  cold adapted, intranasal

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 MICROBIOLOGY
Parvovirus, Papillomavirus, and Polyomavirus
Dr. Palacpac
PARVOVIRUS Erythema Infectiosum (5th Disease)
 Smallest & simplest of all viruses  Children of early school age; occ. Adults
 SS (-) strand DNA; no virion polymerase; icosahedral; naked  Incubation Period: 1-2 weeks
 Virus Protein 2 (VP2) – major capsid protein  Viremia 1 week after infection  persist for 5 days  virus present
 Non Structural Proteins (NS-1) required for replication in nasal washes & gargle specimens
 Highly resistant to inactivation o Pharynx is the most probable site of viral shedding
Since they are naked virus, they are resistant to inactivation by certain  Biphasic  Lytic stage (contagious stage) & immunologic
factors like detergents, soap, pH, etc.  Generalized erythematous rash most important in the face 
“Slapped Cheek” appearance; lace-like rash on limbs or trunk
 Replicated only when a cell is in S phase  Arthralgia (hands & knees) – adults
 Defective virus  requires a helper virus for replication (Herpes virus
or Adenovirus)
In order for them to cause an infection, they would need the help of another
virus such as Herpes virus and Adenovirus

 PaRVo virus
o Human pathogen – B19  with tropism for RBC
progenitors

CLASSIFICATION
1. Parvovirinae
a) Genus Erythrovirus
 Parvovirus B19 (Type 1)
 Strain K71 (Type 2) Recently identified
 Strain V9 (Type 3) human genotypes Other Clinical Syndromes
b) Genus Bocavirus – newly discovered 1. Arthritis in adults – resembles RA (Rheumatoid Arthritis)
c) Genus Dependovirus 2. Pure Red Cell Aplasia
 AAV-2  defective virus  depend on a o Persistent infections and chronic anemia in
helper virus (Adenovirus or Herpes virus) for immunocompromised patients
replication 3. Hydrops Fetalis
2. Densovirinae – infect insects o Result of severe fetal anemia, especially in 2nd trimester
 CHF
B-19 VIRUS  Fetal death before 20th week of pregnancy
 Only member – cause infections in humans o No congenital abnormalities
 Cellular receptor: blood group P antigen (mature erythrocytes,
endothelial cells, placenta, megakaryocytes, fetal liver, heart) Human Bocavirus Respiratory Infection
 Has tropism for:  Prevalent in children with acute wheezing
1) Erythroblasts in Bone Marrow  aplastic anemia  Often found in mixed infections with other viruses
2) Endothelial cells in blood vessels  rashes  Pathogenesis unknown
 MOT:  MOT: probable respiratory route; also detected in stool and serum
1) Respiratory route – primary samples
2) Blood transfusion
3) Vertical/transplacental transmission LABORATORY DIAGNOSIS
 Most sensitive tests detect viral DNA  PCR (most sensitive), in situ
 No evidence of virus excretion in feces or urine hybridization of fixed tissue
 Humans – natural reservoir  Serology:
 Principal targets: immature cells in erythroid lineage (erythroblasts) o B19 IgM antibody – recent infection; present 2-3 months
 Replication  death of infected cells  cessation of RBC production after infection

5th Disease: TREATMENT

 Symptomatic
 Transient aplastic crisis: symptomatic; may require blood transfusion
 Ig preparations containing neutralizing antibodies for
immunocompromised & those with anemia

PREVENTION
 No vaccine
 Good hygiene
 Standard infection control practices for health workers

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 MICROBIOLOGY
Parvovirus, Papillomavirus, and Polyomavirus
Dr. Palacpac
PAPILLOMAVIRUS/POLYOMAVIRUS Picture on Lower Left:
 Causes lytic, chronic, latent & transforming infections depending host They would replicate initially at the basal epithelial cells, whether be in your
cell cervix, uterus, penis, etc. They would first invade the basal cells of these areas
 Small, icosahedral capsid, non-enveloped, dsDNA genomes that would give rise to a latent infection (no virus infection) then later it would
migrate to the upper portions of the skin that would give rise to cell
proliferation and other group of oncogenes directly to the surface of the
1. Early Genes (E1-E7)
epithelium that would cause the assembly of these virus and the release of
o Facilitate replication of viral genome codes for non-
these virus on the surface
structural protein
Modes of Transmission:
2. Late Genes (L1 & L2)
1. Direct contact
o Viral attachment protein
2. Minor skin abrasions
o Codes for structural protein
3. Sexual contact
4. Vertical transmission
HUMAN PAPILLOMAVIRUS (HPV)
 Former members of Papovaviridae Clinical Infections:
 ds circular DNA, icosahedral, naked 1. Skin warts  serotypes 1 to 4
 Oncogenic  E6 & E7 oncoproteins 2. Laryngeal papillomas  serotypes 6 &11
 Do not grow in cell culture 3. Anogenital warts (condylomata acuminata)  6 & 11
 Capsomere form regular pointed star-shaped head 4. Cervical, oropharyngeal & penile CA  16 & 18
 Tissue tropism  epithelial cells of skin & mucous membranes 5. Epidermodysplasia verruciformis
 Encode 7 early genes (E1-E7) & 2 late structural genes (L1 & L2) 
bind with growth suppressor proteins
o E6 protein binds to p53
o E7 protein binds to p105RB
 In permissive cells  cause lytic infections
 In non-permissive cells  abortive, latent, persistent, immortalizing
infections
 Important characteristics:
o Stimulate cell DNA Synthesis
o Significant cause of human cancer
o Viral oncoproteins interact with cellular tumor
suppressor genes

Classification of HPV:
 Alpha papillomavirus
 Beta papillomavirus Laboratory Diagnosis:
 Gamma papillomavirus 1. Cytology
 Mupapapillomavirus 2. DNA molecular probe
 Napapapillomavirus 3. PCR
4. Southern Blot
Pathogenesis:
 Infect & replicate in squamous epithelium of skin (warts) or mucous Treatment:
membrane (papillomas) 1. Removal of the lesion
 Recurrence is common o Surgical excision, Application of caustic agents
 Innate immunity & CMI are important in resolution and control of o Cryosurgery
infections o Electrocautery
o Laser therapy

2. Anti-HPV drugs or immunomodulating drugs

o Imiquimod
o Topical idoxuridine
o Systemic or intralesional alpha interferon
o Cidofovir

Prevention:
 Quadrivalent & Bivalent HPV vaccine – given to 9-54 yrs of age
o Contains virus-like particles composed of HPV L1 proteins
o Quadrivalent – from HPV types 6, 11, 16, and 18
o Bivalent – from HPV types 16 and 18

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 MICROBIOLOGY
Parvovirus, Papillomavirus, and Polyomavirus
Dr. Palacpac
POLYOMAVIRUSES Merkel Cell Polyomavirus (2008)
 Formerly part of Papovaviridae (no longer exists)  share many  Merkel cell carcinomas – skin tumor of neuroendocrine origin
similarities with Papillomaviridae
 Transforming capabilities SV40 Virus
 Circular, dsDNA genome; icosahedral; naked  Fecal-oral route
 Genome with 2 regions:  DNA detected in brain tumors, mesotheliomas, bone tumors &
1) Early region – necessary for replication of viral DNA in lymphomas
permissive cells
2) Late region – for synthesis of coat protein NOTE from Doc Palacpac:
1. Know the morphological differentiation and characteristics
 Stimulate viral oncoproteins interaction with cellular tumor 2. Know the different manifestations
suppressor proteins 3. Know the different serotypes and the carcinomas that they cause
 Important model tumor virus 4. Know the MOT
5. Know the different diseases associated with each virus
 May cause human cancer
REFERENCES
Polyomaviruses:
1. BK Virus  PPT
o Acute hemorrhagic cystitis in bone marrow transplant  Microbiology Manual (2019)
patients & ureteral stenosis  Dr. Palacpac Recordings

2. JC Virus
o Progressive Multifocal Leukoencephalopathy 
destruction of oligodendrocytes; possible role in
colorectal cancer

3. Simian 40 Virus  animal pathogen

BK and JC Viruses
 Usually early childhood
 May persist in kidneys, lymphoid tissues of healthy individuals
 JC virus associated with human brain tumors

KI and WU Viruses
 Discovered in 2007 in nasopharyngeal aspirates from children with
respiratory infections
 Widespread, occur likely in childhood

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“Strength In Knowledge” BESHYWAP 3
 MICROBIOLOGY
ORTHOMYXOVIRUS
Doc Jaime A. Santos | April 23, 2021

Pointers:  The reason why it is highly contagious is because titers peak

 View the recorded lecture
 Read Jawetz
 Read the copy of lecture slides provided in Canvas
OBJECTIVES:
 Describe the general characteristics and structure of
influenza virus and its transmission
 Define what constitutes flu-like illness
 Explain and differentiate antigenic shift and drift and the
nomenclature of influenza A
 Discuss the morbidity and mortality from flu
 Explain how animal flu viruses cause human illness
 Discuss the management and prevention of influenza
INFLUENZA
very early. There is a very short incubation period of only
about 1- 2 days followed by the onset of the respiratory
symptoms.
 Influenza virus spreads from person to person by
airborne droplets or by contact with contaminated hands
or surfaces. A few cells of respiratory epithelium are infected
if deposited virus particles avoid removal by the cough reflex
and escape neutralization by preexisting specific
immunoglobulin A (IgA) antibodies or inactivation by
nonspecific inhibitors in the mucous secretions.
"Flu" symptoms (Flu-like illness)
 Fever/ chills
 Headache
 Nasal congestion
 Sore throat
 Dry cough
 Myalgia/ body aches
 GI symptoms- vomiting/ abdominal pain/ diarrhea
 Malaise/ tiredness
 All of these symptoms can be found in other illnesses and
are nonspecific. That is why they labeled it as flu-like illness.
Other respiratory viruses for example can present this illness.

Clinical Signs and Symptoms

 commonly called "the flu"
 a contagious respiratory illness caused by influenza viruses (Family
Orthomyxoviridae)
 Infection with influenza viruses can result in illness ranging from mild
to severe with life-threatening complications
 incubation period for influenza is 1- 4 days, with an average of 2
days
 Adults - infectious from the day before symptoms begin through
approximately 5 days after onset
 Children - infectious for > 10 days, and young children can shed
virus for < 6 days before their illness onset- signifying long
incubation period
 Immunocompromised persons can shed virus for weeks or
months
 Resolves after a limited number of days for the majority of persons,
although cough and malaise can persist for >2 weeks
 Young children can have initial symptoms mimicking bacterial
sepsis with high fevers _< 20% of children hospitalized with
influenza can have febrile seizures
 Influenza infection has also been associated with:
o Encephalopathy- leading to changes in sensorium
o transverse myelitis- leading to paralysis
o Reye syndrome- px who are taking aspirin at the same time
with flu illness. This can also be seen in association with
aspirin use together with chicken pox
o Myositis- lead to renal failure
o Myocarditis
o Pericarditis

1918 Spanish flu pandemic: 20 to 50 million may have died worldwide

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 MICROBIOLOGY
ORTHOMYXOVIRUS
Doc Jaime A. Santos | April 23, 2021
COVID-19 vs Seasonal Flu  In colder countries flu is largely seen during colder months and they
 Influenza has shorter median incubation period and a shorter vaccinate prior to this season (e.g. October).
serial interval (the time between successive cases) than COVID-  In tropical countries flu is seen all year round.
19 virus. Serial interval for COVID-19 virus is 5-6 days, while for
influenza virus, the serial interval is 3 days
 reproductive number – the number of secondary infections
generated from one infected individual – is 2 and 2.5 for COVID-
19 virus, higher than for influenza
 Children are important drivers of influenza virus transmission in
the community. For COVID-19 virus, data indicate that children
are less affected than adults
 most at risk for severe influenza infection are children, pregnant
women, elderly, those with underlying chronic medical conditions
and those who are immunosuppressed. For COVID-19, older age
and underlying conditions increase the risk for severe infection
 Mortality for COVID-19 appears higher than for seasonal
influenza,
Influenza virus types
 Influenza infection is most commonly associated with
pneumonia. In fact, pneumonia is the most common cause  Three: Influenza A, B, and C
of hospitalization from flu.  Influenza types A or B viruses cause epidemics; influenza A may
cause pandemics
Hospitalization and Deaths  Getting a flu shot can prevent illness from types A and B influenza
 Population at risk for complications, hospitalizations and deaths: but not from type C
o >65 years old  Influenza type C causes mild respiratory illness; not thought to
o young children cause epidemics
o persons of any age with certain underlying health
conditions: cardiovascular and pulmonary (including
asthma), metabolic e.g. DM, Hgbpathies,
immunosuppression
o receiving long term aspirin(ASA)

 Influenza can cause epidemics and pandemics

 In terms of hospital visits, most children have got for consult
compared to adults
 If you look at the hospitalizations from flu, there is 2 peaks,
one in the young and one in the elderly
 In terms of mortality, more deaths among elderly patients,
usually from complications like pneumonia but you may also
have smaller peak in infants.

Seasonal

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 MICROBIOLOGY
ORTHOMYXOVIRUS
Doc Jaime A. Santos | April 23, 2021
 Influenza virus has segmented genome which is very prone
to mutation as well as recombination.
 On the surface, there are spikes and these are hemagglutinin
(HA) and neuraminidase (NA).
 HA is used by the virus for entry into a host cell. They bind to
sialic acid residues on the surface of the host cell.
 The HA protein of influenza virus binds virus particles to
susceptible cells and is the major antigen against which
neutralizing (protective) antibodies are directed. Variability in
HA is primarily responsible for the continual evolution of new
strains and subsequent influenza epidemics. Hemagglutinin
derives its name from its ability to agglutinate erythrocytes
under certain conditions
 NA is important for exit of the virus.
 The NA functions at the end of the viral replication cycle. It is
a sialidase enzyme that removes sialic acid from
glycoconjugates. It facilitates release of virus particles from
infected cell surfaces during the budding process and helps
prevent self-aggregation of virions by removing sialic acid
residues from viral glycoproteins. It is possible that NA helps
the virus negotiate through the mucin layer in the respiratory
tract to reach the target epithelial cells.
 Because of the segmented nature of the genome, when a cell
is coinfected by two different viruses of a given type, mixtures
of parental gene segments may be assembled into progeny
virions. This phenomenon, called genetic reassortment,
may result in sudden changes in viral surface antigens—a
property that explains the epidemiologic features of influenza
and poses significant problems for vaccine development.
 Influenza viruses are relatively hardy in vitro and may be
stored at 0–4°C for weeks without loss of viability. Lipid
solvents, protein denaturants, formaldehyde, and irradiation
destroy infectivity. Both infectivity and hemagglutination are
more resistant to inactivation at alkaline pH than at acid pH.
Replication
 entire Influenza A virus genome is 13,588 bases long and is
contained on eight RNA segments that code for 11 proteins
 RNA synthesis takes place in the cell nucleus, while the synthesis
of proteins takes place in the cytoplasm
 viral proteins are assembled into virions
 the assembled virions leave the nucleus and migrate towards the
cell membrane.
 host cell membrane has patches of viral transmembrane proteins
(HA, NA, and M2) and an underlying layer of the M1 protein which
assist the assembled virions to
NOMENCLATURE
 Influenza A virus divided into subtypes based on HA and N proteins
on surface
 18 HA types, 11 NA types
 Nomenclature based on:
o site of origin
o isolate number
o year of isolation
o subtype
 Example:
 influenzaA/Johannesburg /33/94(H3N2)
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 InfluenzaA/Johannesburg /33/94(H3N2)- this means that
we are dealing with influenza A, first isolated in
Johannesburg, South Africa, isolate number 33, first isolated
1994. In the parenthesis, the HA type is H3 and NA type is
N2.
 Genus Influenzavirus A contains human and animal strains
of influenza type A, Influenzavirus B contains human strains
of type B, and Influenzavirus C contains influenza type C
viruses of humans and swine.
 Antigenic differences exhibited by two of the internal
structural proteins, the nucleocapsid (NP) and matrix (M)
proteins, are used to divide influenza viruses into types A,
B, and C. These proteins possess no cross-reactivity among
the three types.
 Antigenic variations in the surface glycoproteins, HA and NA,
are used to subtype the viruses. Only type A has designated
subtypes.
 The Orthomyxoviridae family also contains the genus
Thogotovirus, members of which are not known to cause
disease in humans
Drift or Shift
 "Antigenic drift" - small changes in the virus that happen
continually every year (influenza A and B).
 Since it occurs yearly, that is the reason why vaccination is
advocated to be done every year.
 Antigenic drift is caused by the accumulation of point
mutations in the gene, resulting in amino acid changes in the
protein. Sequence changes can alter antigenic sites on the
molecule such that a virion can escape recognition by the
host’s immune system. The immune system does not cause
the antigenic variation but rather functions as a selection
force that allows new antigenic variants to expand. A variant
must sustain two or more mutations before a new,
epidemiologically significant strain emerges
 "Antigenic shift" - abrupt, major change in the influenza A viruses,
resulting in new H &/or new H and N proteins that infect humans
(influenza A only).
 Influenza A can cause pandemics.
 Antigenic shift reflects drastic changes in the sequence of
a viral surface protein, changes too extreme to be explained
by mutation. The segmented genomes of influenza viruses
reassort readily in doubly infected cells. The mechanism for
shift is genetic reassortment between human and avian
influenza viruses. Influenza B and C viruses do not exhibit
antigenic shift because few related viruses exist in animals.
 All three types of influenza virus exhibit antigenic drift.
However, only influenza A undergoes antigenic shift,
presumably because types B and C are restricted to
humans, but related influenza A viruses circulate in animal
and bird populations. These animal strains account for
antigenic shift by genetic reassortment of the glycoprotein
genes. Influenza A viruses have been recovered from many
aquatic birds, especially ducks; from domestic poultry, such
as turkeys, chickens, geese, and ducks; from pigs and
horses; and even from seals and whales. Serologic surveys
indicate a high prevalence of influenza virus infection in
domestic cats.
Subtypes in humans
 Current subtypes of influenza A viruses found in people are A(H1N1)
and A(H3N2)
 MICROBIOLOGY
ORTHOMYXOVIRUS
Doc Jaime A. Santos | April 23, 2021
 Influenza B virus is not divided into subtypes.
 Influenza A(H1N1), A(H3N2), and influenza B strains are
included in each year's influenza vaccine
 Protection against flu is serotype specific, which is one reason why
you can get flu every year.
 In 1980, Influenza B virus diverge into 2 lineages: the
Victoria lineage and the Yamagata lineage.
 The period between epidemic waves of influenza A tends to
be 2–3 years; the interepidemic period for type B is longer
(3–6 years). Every 10–40 years, when a new subtype of
influenza A appears, a pandemic result. This happened in
1918 (H1N1), 1957 (H2N2), and 1968 (H3N2). The H1N1
subtype reemerged in 1977, although no epidemic
materialized. Since 1977, influenza A (H1N1) and (H3N2)
viruses and influenza B viruses have been in global
circulation.
 H1, H2, H3 and the other H types are found in animals and
humans influenza viruses.
 N1 and N2 are found in human influenza virus, together with
H1, H2 and H3 only.
 For example, an Influenza A(H5N1)- you can be sure that its
an animal flu virus because H1, H2, H3 and N1 and N2 are
found in human flu viruses only.
 Chicken, pigs, and birds have their own flu viruses. However,
humans can get avian flu upon exposure to chicken. They
can also get swine flu from pigs.
 Animal viruses are not very good at infecting humans,
requiring very close contact between humans and animals.
 If a pig have both human and bird flu, there is opportunity for
recombination and because human flu can be combined with
bird flu or animal flu virus, when transmitted back to humans
from the mixing vessels there can be significant human to
human transmission. This can result to antigenic shift and
what is happens subsequently will be Influenza pandemics.
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Most recent Flu Pandemic: Novel A(H1N1) •
 quádruple reassortant : 2 NA bird flu genes, 1 swine flu, human flu
 thrives in lower respiratory tract
 more contagious


has now become seasonal flu
resistant to Amantadine, Rimantadine
Avian Influenza
 Many subtypes of influenza A virus subtype of avian influenza
 Low pathogenicity avian influenza (LPAI)
o Causing lower mortality among birds or chickens
 Highly pathogenic avian influenza (HPAÍ)
o First recognized in Italy in 1878
o Extremely contagious in birds
o Rapidly fatal, high mortality (almost 100% În a few days)
o Documented avian flu in humans:
 H5N1, H7N3, H7N7, H7N9, and H9N2, H10N8
Treatment of Influenza
 Antiviral Medications:
o antiviral drugs: adamantanes: Amantadine, Rimantadine-
not used anymore because of resistance
o neuraminidase inhibitors: Zanamivir, Oseltamivir- newer
drugs
o Antiviral treatment lasts for 5 days and must be started
within the first 2 days of illness.
Flu vaccination
 Inactivated vaccines
o the whole virus vaccine, split virus vaccine (disrupted by
detergent, and the most common vaccine), and subunit
vaccine (H and N purified).
o Some formulations include adjuvants to enhanced
immunogenicity.
o The one which are available, intramuscular or
subcutaneous route
 Live, attenuated influenza vaccines
o have been based on a temperature-sensitive variant
vaccine virus strains that replicate well in the nasopharynx
but poorly in the lower respiratory tract.
o Not indicated for children and for immunocompromised
person because they are lived virus.
o More expensive, intranasal route and not available locally
When Should You Give the Flu Shot?
 Highlights Influenza Consensus – Flu TWG Oct. 2004
 Data from the five-year epidemiologic data (February 1998 –
September 2003) shows increased influenza activity June to
November.
 Vaccination should be given once a year preferably from February
to June
 The Southern Hemisphere vaccine which is made available
starting February of each year is recommended to cover the
expected increase in influenza activity from June to November.
 MICROBIOLOGY
ORTHOMYXOVIRUS
Doc Jaime A. Santos | April 23, 2021

References
 PPT and Recording
 Jawetz

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