Chapter 43

1
 Human cancer viruses

 At least 15-20% of all human tumors worldwide have a viral

cause
 two of great significance worldwide - cervical cancer and liver cancer

 viruses strongly associated with human cancers:

 human papillomaviruses
 Epstein-Barr virus
 human herpesvirus 8
 hepatitis B virus
 hepatitis C virus
 Merkel cell virus
 two human retroviruses (HIV, HTLV)
Human T-lymphotropic virus

 Tumor viruses can produce tumors when they infect appropriate

animals
2
 TABLE 43-1
Association of Viruses
With Human Cancers

There are 7 known human

tumor-associated viruses, 2 –
RNA viruses, 5 – DNA viruses
 Human Immunodeficiency

Virus – HIV
 cause of acquired immune

deficiency syndrome - AIDS

 cytolytic and

nontransforming
 But the immunity ↓ → ↑ risk

of several types of cancer:

 cervical cancer, Kaposi

sarcoma, lymphomas, head

and neck cancer, liver
cancer, and oral cancer
3
 TABLE 43-2 Tenets of Viral Carcinogenesis
 1. Viruses can cause cancer in animals and humans
 2. Viruses are seldom complete carcinogens
 3. Host factors are important determinants of virus-induced tumorigenesis

 4. Tumor viruses frequently establish persistent infections in natural hosts

 5. Long latent periods usually elapse between initial virus infection and tumor
appearance

 6. Viral strains may differ in oncogenic potential

 7. Viruses may be either direct- or indirect-acting carcinogenic agents
 8. Oncogenic viruses modulate growth control pathways in cells
 9. Animal models may reveal mechanisms of viral carcinogenesis
 10. Viral markers are usually present in tumor cells
 11. One virus may be associated with more than one type of tumor

 12. Virus infections are more common than virus-related tumor formation
4
 Mechanisms of Action by Human Cancer Viruses

 2 general patterns :
 tumor virus introduces a new "transforming gene" into the cell (direct-acting)
or
 virus alters the expression of a preexisting cellular gene or genes (indirect-
acting)
 In either case, the cell loses control of normal regulation of growth processes –
becomes transformed

 Transformation - cells' acquisition of some growth property not exhibited by

the parental cell type
 Transformation to a malignant phenotype is recognized by tumor
formation when transformed cells are injected into appropriate test
animals

 Indirect-acting tumor viruses are not able to transform cells in culture

5
 Tumor Viruses
3 mechanisms of tumor formation
highly oncogenic weakly oncogenic
RNA viruses RNA viruses DNA viruses
(mostly defective)

Virus carries onc gene Virus enhances Virus inactivates cellular

cellular oncogenes tumor suppressors genes
Directly-acting Indirectly-acting

Not in humans Human T-lymphotrophic Human papilloma virus, HPV

Virus, HTLV

onc gene of
Ab-MLV, Abelson murine leukemia virus
cellular origin
(abl oncogene) (Gammaretrovirus)

Ab-MLV - defective

RSV, Rous sarcoma virus (src

oncogene) (Alpharetrovirus)

Exception: RSV -
Lentiviruses contain 4 genes 6
nondefective required for a replicating retrovirus
— gag, pro, pol, and env
 Tumor Viruses Are of Different Types
Most tumor viruses either
 have a DNA genome or
 have a RNA genome and generate a DNA provirus after infection of cells
(hepatitis C virus is an exception)

 DNA tumor viruses

 classified among the papilloma-, polyoma-, adeno-, herpes-, hepadna-, and
poxvirus groups
 encode viral oncoproteins - important for viral replication
 that also affect cellular growth control pathways

 RNA tumor viruses

 most belong to the retrovirus family
 highly oncogenic (direct-transforming) viruses carry an oncogene of cellular

origin
• presence of the cellular sequences is of no benefit to the viruses
 weakly oncogenic (slowly transforming) viruses do not contain an oncogene

and induce leukemias after long incubation periods by indirect mechanisms

7
 Multistep Carcinogenesis

 multiple genetic changes - from 5 to 8 - must occur to convert a normal cell

into a malignant one

 Intermediate stages are designated by terms such as "immortalized,"

"hyperplasia," and "preneoplastic“

 Tumors usually develop slowly over a long period of time

 history of cancers suggests repeated selection of rare cells with some

selective growth advantage

 activation of multiple cellular oncogenes and inactivation of tumor

suppressor genes - involved in the evolution of tumors whether or not a
virus is involved

 tumor virus usually acts as a initiators of the neoplastic process

8
 Multistep Carcinogenesis

 Effect of cellular oncogenes and tumor suppressor genes on the cell

cycle:
 Oncoproteins encoded by cellular oncogenes activate the cell cycle by
allowing passage from the G1 phase into the S phase.
 Proteins encoded by tumor suppressor genes (negative regulators of cell
growth), notably p53 and RB, inhibit the cell cycle in the G1 phase -
Inactivation of these proteins activates the cell cycle by allowing passage
from the G1 phase into the S phase.
• G1, gap 1; G2, gap 2; M, mitosis; S, synthesis of DNA

 p53 gene also

causes cells with
DNA damage to
undergo apoptosis

 loss of p53 function allows cells

with damaged DNA to progress
through the cell cycle, leading9 to
the accumulation of genetic
 Tumor Suppressor Genes

 Examples of Tumor Suppressor Genes Involved in Human Cancer

 in many colon carcinomas, two genes are inactivated, the p53 gene
and the DCC (deleted in colon carcinoma) gene

 >1/2 of human cancers have a mutated p53 gene in the DNA of

malignant cells 10
 Cellular Oncogenes-1
 “Oncogene” - gene involved in cancer causation

 Normal precursors of the cellular oncogenes are proto-oncogenes

 they encode normal cellular proteins and are under regulatory control

 Cellular oncogenes have acquired mutations that cause them to escape

regulatory control and overproduce altered proteins

 Cellular oncogenes may be the precursors of viral oncogenes

 cellular oncogenes have exons and introns, whereas viral oncogenes do not

 Studies with acutely transforming nonhuman retroviruses revealed:

 During virus replication - provirus stage - cellular sequences (oncogenes)
had been captured and incorporated into the retrovirus genomes
Retroviruses - transducing agents,
carrying oncogenes from one cell to another
 presence of the cellular sequences is of no benefit to the viruses
11
 Cellular Oncogenes-2

 two different mechanisms - mutation and increased expression – activate

the quiescent “proto-oncogene” into a functioning oncogene capable of
transforming a cell:

I. Mutation - genetic damage – the gene may be:

 overexpressed → overproduced oncogene product may be important in
cellular growth changes
 expressed at the wrong time during the cell cycle
 expressed in inappropriate tissue types
Indirectly-acting
RNA viruses

II. Insertion of a retroviral promoter adjacent to a cellular oncogene may result

in enhanced expression of that gene - “promoter-insertion
oncogenesis”

12
 Interactions of Tumor Viruses with Their Hosts
 The known tumor viruses establish long-term persistent infections in
humans

 So they must avoid detection and recognition by the immune system that
would eliminate the infection

 Different viral evasion strategies exist:

 restricted expression of viral genes that makes infected cells nearly invisible to
the host (EBV in B cells)
 infection of sites relatively inaccessible to immune responses (HPV in the
epidermis)
 mutation of viral antigens that allows escape from antibody and T cell
recognition (HIV)
 modulation of host MHC class I molecules in infected cells (adenovirus,
cytomegalovirus) - the infected cell can no longer be recognized by cytotoxic T
cells
 inhibition of antigen processing (EBV)
 infection and suppression of essential immune cells (HIV) 13
 Cell Susceptibility to Viral Infections & Transformation
 Host cells are either permissive or nonpermissive for replication of a given
virus:
 permissive cells support viral growth and production of progeny virus
 nonpermissive cells do not
 DNA viruses:
 permissive cells are often killed by virus replication and are not transformed
unless viral replicative cycle is blocked in some way
 nonpermissive cells may be transformed

 A characteristic property of RNA tumor viruses - they are not lethal for the cells
in which they replicate

 Most tumor viruses exhibit marked tissue specificity:

 variable presence of surface receptors for the virus
 ability of the virus to cause disseminated versus local infections

 Some viruses are associated with a single tumor type, whereas others are linked
to multiple tumor types – depends on tissue tropisms of the viruses
14
 Retention of Tumor Virus Nucleic Acid in a Host Cell

 The stable genetic change from a normal to a neoplastic cell generally requires
the retention of viral genes in the cell

retention of viral genes in the cell:

 DNA tumor viruses - integration of certain viral genes into the host cell genome
 episomal copies of the viral genome are maintained in tumor cells
 proviral DNA copy of the viral RNA is integrated in the host cell DNA
(retroviruses)
 genome RNA copies of hepatitis C virus are maintained in tumor cells

 In some viral systems, virus-transformed cells may release growth factors that
affect the phenotype of neighboring uninfected cells, thereby contributing to
tumor formation

 It is also possible that as tumor cells collect genetic mutations during tumor
growth, the need for the viral genes that drove tumor initiation may become
unnecessary and viral markers will be lost from some transformed cells
15
 RNA TUMOR VIRUSES
HEPATITIS C VIRUS

 member of the Flaviviridae family

 55-65 nm enveloped virions
 (+) sense single-stranded RNA 9.4 kb

 It appears that the majority of infections

become persistent, even in adults

 Chronic infection with hepatitis C virus

→ chronic inflammation and cirrhosis →
hepatocellular carcinoma

 There are over 170 million chronic

carriers of hepatitis C virus
 1-5% of these going on to develop
hepatocellular carcinoma 16
 RNA TUMOR VIRUSES
Retroviruses
 contain
 two identical subunits of (+) sense ssRNA
 RNA-directed DNA polymerase (reverse transcriptase) - inside virions

 Replication: reverse transcriptase makes DNA copy from genomic RNA; DNA
(provirus) integrates into cellular chromosome; provirus is template for viral RNA

 Infections do not kill cells - exceptions are the lentiviruses, may be cytolytic
 Proviruses remain permanently associated with cells and are frequently not
expressed
 Many members are tumor viruses Lentiviruses
 mainly cause tumors of
 reticuloendothelial and hematopoietic systems
(leukemias, lymphomas)
 connective tissue (sarcomas)

 Retroviruses - isolated from virtually all vertebrate species

17
 Retroviruses - Classification
7 genera

Retroviridae

Epsilonretrovirus Spumavirus
(fish viruses) (no known disease)

Gammaretrovirus Deltaretrovirus
Alpharetrovirus Betaretrovirus (HTLV)

avian leukosis and human T-lymphotropic

sarcoma viruses viruses and bovine
Lentivirus leukemia virus
mouse mammary (HIV)
tumor virus mammalian leukemia
and sarcoma viruses 18
 RNA TUMOR VIRUSES
Retroviruses

 spherical, 80–110 nm in diameter, helical

ribonucleoprotein within icosahedral
capsid, enveloped

 ssRNA, linear, (+) sense, 7–11 kb, diploid;

may be defective; may carry oncogene

 Maturation: Virions bud from plasma

membrane

 type-specific or subgroup-specific
antigens are associated with the
glycoproteins in the viral envelope -
encoded by the env gene

 group-specific antigens are associated

with the virion core - encoded by the gag
gene - shared by viruses from the same
host species
19

Gammaretroviruses
 RNA TUMOR VIRUSES
Retroviruses
morphologic classes based on EM
 3 extracellular morphologic classes – B, C, D and intracellular – A types
 They reflect slightly different processes of morphogenesis by different
retroviruses

 Type A particles occur only intracellularly and appear to be noninfectious:

 intracytoplasmic type A particles, 75 nm in diameter, are precursors of
extracellular type B and D viruses
 intracisternal type A particles, 60–90 nm in diameter, are unknown entities

 Type B viruses are 100–130 nm in diameter and contain an eccentric nucleoid

 The type C viruses represent the largest group of retroviruses:

 The particles are 90–110 nm in diameter, and the nucleoids are centrally located
 The type C viruses may exist as exogenous or endogenous entities

the vast majority of isolates

 The type D retroviruses are poorly characterized: display type C characteristics
 The particles are 100–120 nm in diameter, contain an eccentric nucleoid, and
exhibit surface spikes shorter than those on type B particles
20
 RNA TUMOR VIRUSES
Retroviruses
morphologic classes based on EM
 Comparative morphology of types A, B, C, and D retroviruses. A:
Intracytoplasmic type A particles (representing immature precursor of budding
type B virus). B: Budding type B virus. C: Mature, extracellular type B virus.

 Type A particles occur only intracellularly and appear to be noninfectious:

 intracytoplasmic type A particles - 75 nm in diameter - precursors of extracellular
type B and D viruses
 intracisternal type A particles, 60–90 nm in diameter, are unknown entities
 Type B viruses are 100–130 nm in diameter and contain an eccentric nucleoid

21
 RNA TUMOR VIRUSES
Retroviruses
morphologic classes based on EM

 Comparative morphology of types A, B, C, and D retroviruses. D: Lack of

morphologically recognizable intracytoplasmic form for type C virus. E: Budding
type C virus. F: Mature, extracellular type C virus.

 The type C viruses represent the largest group of retroviruses.

 The particles are 90–110 nm in diameter, and the nucleoids are centrally located
 The type C viruses may exist as exogenous or endogenous entities

22
 RNA TUMOR VIRUSES
Retroviruses
morphologic classes based on EM

 Comparative morphology of types A, B, C, and D retroviruses. G:

Intracytoplasmic type A particle (representing immature precursor form of type D
virus). H: Budding type D virus. I: Mature, extracellular type D virus

 The type D retroviruses are poorly characterized.

 The particles are 100–120 nm in diameter, contain an eccentric nucleoid, and
exhibit surface spikes shorter than those on type B particles

23
 RNA TUMOR VIRUSES Viral genetic information that is a
constant part of the genetic
Retroviruses constitution of an organism is
Exogenous or Endogenous designated as “endogenous”
 Exogenous  Endogenous

 spread horizontally  spread vertically through the germ line

 behave as typical infectious agents  do not produce any disease and
 initiate infection and transformation cannot transform cells in culture
only after contact
 The pathogenic retroviruses all
 usually not pathogenic for their host
appear to be exogenous viruses animals
 found only in infected cells
 found in all cells (somatic and germ) -
DNA copies of RNA tumor virus
genomes are covalently linked to
exception: There are examples of disease cellular DNA
caused by replication of endogenous  Many vertebrates, including humans,
viruses in inbred strains of mice possess multiple copies of
endogenous RNA viral sequences
 may be induced to replicate either
integrated retroviral provirus behaves spontaneously or by treatment with
like a cluster of cellular genes and is extrinsic (chemical) factors
subject to regulatory control by the cell –
partial or complete repression of viral
24
gene expression
 Retroviruses
mouse mammary tumor virus

 Morphologic type B viruses: 100–130 nm, contain an eccentric nucleoid

 The prototype of this group - mouse mammary tumor virus, Betaretrovirus genus
 occurs in “high mammary cancer” strains of inbred mice
 found in particularly large amounts in lactating mammary tissue and milk
 readily transferred to suckling mice → in whom the incidence of subsequent
development of adenocarcinoma of the breast is high

 Many vertebrates possess multiple copies of endogenous RNA viral sequences -

of no apparent benefit to the animal
 However, endogenous proviruses of mammary tumor virus carried by inbred
strains of mice express superantigen activities that influence the T-cell
repertoires of the animals

25
 Retroviruses
Host Range
 Natural infections by a given virus are usually limited to a single species, though
infections across species barriers may occur

 The presence or absence of an appropriate cell surface receptor:

 a major determinant of the host range of a retrovirus
 Infection is initiated by an interaction between the viral envelope glycoprotein
and a cell surface receptor

 Ecotropic viruses infect and replicate only in cells from animals of the original
host species

 Amphotropic viruses exhibit a broad host range (able to infect cells not only of
the natural host but of heterologous species as well) because they recognize a
receptor that is widely distributed

 Xenotropic viruses can replicate in some heterologous (foreign) cells but not in
cells of the natural host
 Many endogenous viruses have xenotropic host ranges
26
 Retroviruses
Genetic Content I

 Retroviruses have a simple genetic content, but there is some variation in the
number and type of genes contained
 The genetic makeup of a virus influences its biologic properties
 Genomic structure - a useful way of categorizing RNA tumor viruses
 The gene order in all retroviruses is 5'-gag-pro-pol-env-3'

Alpharetrovirus and Gammaretrovirus

MA – matrix associated
protein, SU – surface
CA – capsid protein, glycoprotein (spike),
NC - nucleocapsid protein TM – transmembrane
glycoprotein (spike)

27
 Retroviruses
Genetic Content II
 The gene order in all retroviruses is 5'-gag-pro-pol-env-3‘:
 gag - encodes core proteins (group-specific antigens)
 pro - encodes a protease enzyme
 pol - encodes the reverse transcriptase enzyme (polymerase)
 env - encodes glycoproteins that form projections on the envelope
Standard leukemia viruses –
Alpharetrovirus and Gammaretrovirus – simple genome

28
 Retroviruses
Genetic Content III
 human retroviruses (Deltaretrovirus and Lentivirus), contain additional genes –
complex genome - downstream from the env gene (tax or tat - transactivating
regulatory gene - encodes a nonstructural protein that alters the transcription or
translational efficiency of other viral genes)
 Transactivation - increased rate of gene expression triggered by
transactivators
 Retroviruses with either of these two genomic structures (simple or complex) will
be nondefective, replication-competent (in appropriate cells)
 Because they lack a transforming (onc) gene, they cannot transform cells in
tissue culture - However, they may have the ability to transform precursor cells in
blood-forming tissues in vivo

Simple genome
MLV, murine leukemia virus
(Gammaretrovirus)

Complex genome
HTLV, human T-
lymphotropic virus
29
(Deltaretrovirus)
 Retroviruses
Genetic Content III
 The directly transforming retroviruses carry an onc gene
 The transforming genes represent cellular genes that have been appropriated by
those viruses at some time in the distant past and incorporated into their
genomes

 addition of the cellular DNA almost always results in the loss of portions of the
viral genome
 Consequently, the sarcoma viruses usually are replication-defective; progeny
virus is produced only in the presence of helper viruses (leukemia viruses)
 Such viruses are highly oncogenic in appropriate host animals and can
transform cells in culture

Mo-MSV, Moloney murine

sarcoma virus (mos oncogene)
(Gammaretrovirus)
replication -defective

MLV, murine leukemia virus

(Gammaretrovirus)
30
Helper virus - nondefective
 Retroviruses
Oncogenic Potential

 neoplastic transformation by retroviruses - the result of a cellular gene that is

normally expressed at low, carefully regulated levels becoming activated and
expressed constitutively:

 In the case of the acute transforming viruses - retroviruses that contain

oncogenes - a cellular gene has been inserted by recombination into the viral
genome and is expressed as a viral gene under the control of the viral promoter
 they induce tumors in vivo after very short latent periods and rapidly induce morphologic
transformation of cells in vitro

 In the case of the slow-transforming leukemia viruses - that do not carry an

oncogene have a much lower oncogenic potential - the viral promoter or
enhancer element is inserted adjacent to or near the cellular gene in the
cellular chromosome
 disease (usually of blood cells) appears after a long latent period; cultured cells are not
transformed
31
 Retroviruses Deltaretrovirus
Replication of Retroviruses-1
Human T-Lymphotropic Virus
 virus attaches to cell surface receptor → viral
capsid enters the cell
 viral reverse transcriptase - functioning as an
RNA-dependent DNA polymerase - produces
DNA copy of genome RNA within capsid in
cytoplasm
proviral DNA is
 DNA enters nucleus → integrated at random transcribed by the host
into cell DNA, forming provirus RNA polymerase II
 integrated provirus serves as template for
synthesis of viral transcripts, some - are
unspliced and will be encapsidated as genomic
RNAs and some - are spliced and will serve as
mRNAs
 Viral proteins are synthesized → proteins and
genome RNAs assemble→particles bud from cell
 Capsid proteins are proteolytically processed by
viral protease → producing mature, infectious
virions - shown schematically as conversion from
a square to icosahedral core 32
 Retroviruses
Replication of Retroviruses-2

 By a complex process, sequences from both ends of the viral RNA become
duplicated, forming the long terminal repeat located at each end of the viral DNA
 Long terminal repeats are present only in proviral DNA

 Comparison of structures of retrovirus RNA genome and integrated provirus

DNA
 A virus particle contains 2 identical copies of the ssRNA genome. The 5′ terminal is
capped, and the 3′ terminal is polyadenylated. A short sequence, R, is repeated at
both ends; unique sequences are located near the 5′ (U5) and 3′ (U3) ends.
U3 contains promoter and enhancer sequences.
 The integrated provirus DNA is flanked at each end by the long terminal repeat (LTR)
structure generated during synthesis of the DNA copy by reverse transcription.
33
Each LTR contains U3, R, and U5 sequences.
 Retroviruses
Replication of Retroviruses-3

 structure of the provirus is constant

 its integration into host cell genomes can occur at different sites
 The very precise orientation of the provirus after integration is achieved by
specific sequences at the ends of both long terminal repeats

 Provirus remains integrated within the cellular DNA for the life of the cell

 There is no known way to cure a cell of a chronic retrovirus infection

 If the RNA virus happens to contain a transforming gene, the oncogene plays no
role in replication

 This is in marked contrast to the DNA tumor viruses, in which the transforming
genes are also essential viral replication genes

34
 Deltaretrovirus
Human Retroviruses
Human T-Lymphotropic Viruses -a
 HTLV-1 causes
 adult T cell leukemia/lymphomas (ATL)
 HTLV-1–associated myelopathy/tropical spastic paraparesis (HAM/TSP) -

nervous system degenerative disorder

 uveitis
 infective dermatitis of the same magnitude
progressive weakness of the
and importance in
legs and lower body; patient’s
mental faculties remain intact;
tropics as is multiple
chronic, severe and recurrent
usually appears in 2-3 years sclerosis in Western
eczema occurring during
after infection childhood in patients vertically countries
infected with HTLV-1
 3 related human viruses - HTLV-2, HTLV-3, and HTLV-4 – have not been
conclusively associated with a specific disease
 HTLV-1 and HTLV-2 share about 65% sequence homology and display
significant serologic cross-reactivity

 human lymphotropic viruses have a marked affinity for mature T cells

35
 Human Retroviruses
Human T-Lymphotropic Viruses -b
 HTLV does not possess an oncogene in its genome and does not integrate
its proviral DNA at a specific site near a cellular oncogene in the T-cell DNA
(i.e., it does not cause insertional mutagenesis)
 transregulating human retroviruses:
 carry a gene, tax, whose product - nonstructural protein - alters the
expression of other viral genes - transcription or translational efficiency
 Transactivating regulatory genes –
 necessary for viral replication in vivo
 may contribute to oncogenesis by also modulating cellular genes that

regulate cell growth

36
 Human Retroviruses
Human T-Lymphotropic Viruses -c
 HTLV-1 has 6 reported subtypes (subtypes A to F) - the major ones being
subtypes A, B, and C
 distributed worldwide - 20 million infected individuals
 Clusters of HTLV-associated disease are found in certain geographic areas
(southern Japan, Melanesia, the Caribbean, Central and South America, and
parts of Africa)
 up to 5% of the population in endemic areas may be sero(+)
 ATL (adult T cell leukemia) typically appears 20 to 30 years after initial infection
 ATL is poorly responsive to therapy - 5-year survival rate for patients with this
cancer is <5%
Such early-life infections
cell-associated virus via are associated with the
infected lymphocytes greatest risk of ATL
Transmission:

 breastfeeding - an important mode - efficiency of transmission 15-20%
 blood transfusion screening donated blood
 blood-contaminated needles (drug abusers) for antibodies to HTLV
 sexual intercourse (infected lymphocytes in semen)
37
 Human Retroviruses
Human T-Lymphotropic Viruses -d
 In the blood, the malignant T cells have a distinct “flower-shaped” nucleus:

Typical “cloverleaf” appearance of

nuclei of HTLV-1–infected adult
T-cell leukemic cells.

HTLV-1 infection both -

stimulates mitosis and
immortalizes T lymphocytes
 HTLV primarily infects CD4-positive T lymphocytes
 HTLV-encoded Tax protein ↑ synthesis of IL-2 receptor and IL-2 (T-cell growth
factor) → ↑ rapid T-cell growth and eventually its malignant transformation
 HTLV remains latent within the malignant T cells - HTLV is typically not produced
by the malignant cells

 HAM - a demyelinating disease of the brain and spinal cord, especially of the
motor neurons in the spinal cord - caused either by an autoimmune cross-
reaction in which the immune response against HTLV damages the neurons or38 by
cytotoxic T cells that kill HTLV-infected lymphocytes infiltrated int he CNS
 Retroviruses
Human Retroviruses
 .
 Human Immunodeficiency Viruses - HIVs
 classified as lentiviruses
 cause of acquired immune deficiency syndrome - AIDS
 cytolytic and nontransforming
 But the immunity ↓ → ↑ risk of several types of cancer:
 cervical cancer, Kaposi sarcoma, lymphomas, head and neck cancer, liver
cancer, and oral cancer

 Other retroviruses
 simian foamy viruses (Spumavirus genus)
 able to cause “foamy” degeneration of inoculated cells - not associated with any
known disease process
 highly prevalent in captive nonhuman primates
 humans occupationally exposed to the primates can be infected - but these
infections have not resulted in any recognized disease
39
 RNA and DNA Tumor Viruses

 Fundamental differences exist between the oncogenes of DNA and RNA tumor
viruses

 Transforming genes carried by DNA tumor viruses encode functions

required for viral replication and do not have normal homologs in cells

 In contrast, RNA tumor viruses:

 either carry transduced cellular oncogenes with no role in viral
replication
 or they act through indirect mechanisms

 The DNA virus transforming proteins complex with normal cell proteins
and alter their function

 To understand the mechanism of action of DNA virus transforming proteins, it

is important to identify the cellular targets with which they interact 40
 DNA Tumor Viruses
 TABLE 43-4 Examples of DNA Virus Oncoproteins and Cellular Protein
Interactions

p53, product of p53 gene; pRb, retinoblastoma gene product; PP2A, protein
phosphatase 2A; DLG, MAGI-1, and MUPP1, members of a family of cellular
proteins that contain PDZ domains; PDGF, platelet-derived growth factor; EBV,
Epstein-Barr virus; LMP (latent membrane protein) TRAF, tumor necrosis factor 41

receptor–associated factor
 DNA Tumor Viruses
Polyomaviruses
 Family – Polyomaviridae*, single genus designated Polyomavirus
 Small, icosahedral, 45 nm in diameter, nonenveloped
 circular dsDNA, 5 kbp, cellular histones condense DNA in virion
 Replication in nucleus
 Outstanding characteristics:
 Stimulate cell DNA synthesis
 Viral oncoproteins interact with cellular tumor suppressor proteins
 Important model tumor viruses
 Human viruses can cause human neurologic and renal disease
 May cause human cancer

Three structural (capsid)

proteins - VP1, VP2, VP3

42

*Formerly classified in Papovaviridae family

 DNA Tumor Viruses
Polyomaviruses

 The name polyoma refers to the viruses' ability to produce multiple (poly-)
tumors (-oma)

 simple DNA-containing viruses:

 possess a limited amount of genetic information (6 or 7 genes)

 Multiple species have been identified:

 tumor virus SV40
 others known to infect humans - BK, JC, KI, WU, MCV, HPyV6, HPyV7,
HPyV10, and TSV

 Many species of mammals and some birds have been found to carry their
own species of polyomavirus

43
DNA Tumor Viruses
Polyomavirus SV40

44
 DNA Tumor Viruses
Polyomavirus Replication

 genome contains "early" and "late" regions:

 The early region is expressed soon after infection of cells:
contains genes that code for early proteins—eg, the SV40 large tumor (T)
antigen, which is necessary for the replication of viral DNA in permissive cells,
and the small tumor (t) antigen

 The late region - genes that code for the synthesis of coat protein; they have
no role in transformation and usually are not expressed in transformed cells
abolishing its function would lead to accumulation of T
antigen-expressing cells with genomic mutations
 T antigen interacts with the cellular tumor suppressor gene products, p53
and pRb family members (inactivates them) - allowing cells to enter S phase
so that viral DNA may be replicated

 The functional inactivation of the cellular proteins by T antigen binding -

central to the virus-mediated transformation process
45
 Polyomavirus
Pathogenesis & Pathology I

 human polyomaviruses (BK and JC) - widely distributed - the presence of specific
antibody in 70–80% of adult sera
 Infection usually occurs during early childhood

 Both viruses may persist in the kidneys and lymphoid tissues of healthy
individuals after primary infection

 reactivate when the host's immune response is impaired, eg,

 by renal transplantation
 during pregnancy
 increasing age
 Viral reactivation and shedding in urine are asymptomatic in
immunocompetent persons

 These two viruses - antigenically distinct - but both encode a T antigen that is
related to SV40 T antigen
46
 DNA Tumor Viruses
Polyomavirus
Pathogenesis & Pathology II

 BK virus causes
 hemorrhagic cystitis in bone marrow transplant recipients
 polyomavirus-associated nephropathy in up to 5% renal transplant recipients -
graft failure in up to 50% of those affected patients

 JC virus
 the cause of progressive multifocal leukoencephalopathy (PML) - a fatal brain
disease that occurs in some immunocompromised persons, especially with
depressed cell-mediated immunity

 acquired by the respiratory route (droplets) and spread by viremia to establish

latent infection in multiple organs

 JC virus has been associated with human brain tumors, but an etiologic role is not
yet established
47
 Slow Virus Infections
Progressive Multifocal Leukoencephalopathy

 JC virus (JCV), a member of

the family Polyomaviridae

 In immunosuppressed
individuals, JCV is activated,
spreads to the
brain→oligodendrocyte
infection → CNS
demyelination

 coma and death within 6

months of onset

 Once exceedingly rare, the

disease is now seen in a
significant proportion (about
5%) of patients with AIDS
48
 DNA Tumor Viruses
Polyomavirus
Pathogenesis & Pathology III

 KI and WU viruses - discovered in 2007 in nasopharyngeal aspirates from

children with respiratory infections
 Merkel cell polyomavirus - identified in 2008 in Merkel cell carcinomas - rare
skin tumors of neuroendocrine origin
 Seroprevalence studies suggest that KI, WU, and Merkel cell virus infections
are widespread and likely occur in childhood

 Merkel cell carcinomas

 In carcinoma cells, the DNA of MCPV is integrated into cell DNA
 gene for the large T antigen is mutated so the virus cannot replicate but the T
antigen continues to be synthesized
 T antigen causes the cell to become malignant by inhibiting tumor suppressor
proteins such as p53 and RB
 Because MCPV does not replicate in the carcinoma cells, patients are not
infectious to others
49
 DNA Tumor Viruses
Polyomavirus
Pathogenesis & Pathology IV

 Merkel cell carcinomas, rare skin tumors of neuroendocrine origin

early diagnosis and treatment of Merkel cell cancer are important

factors in decreasing the chance of its spreading, after which it is
50
difficult to cure
 DNA Tumor Viruses
Polyomavirus
Pathogenesis & Pathology V

 HPyV6, HPyV7, and HPyV10 appear to be constituents of human skin

 Trichodysplasia spinulosa–associated polyomavirus (TSV) was discovered

in proliferative skin lesions

 HPyV9 was found in the blood of immunosuppressed patients

 HPyV12 was found in liver tissue

 Other polyomaviruses have been found in human stool - MWPyV, MXPyV, and
STL polyomavirus

 Because of their recent discoveries, information on disease associations is

limited
51
 DNA Tumor Viruses
Polyomavirus Tumor induction in the
Pathogenesis & Pathology VI natural host - rhesus
monkey - is rarely
observed
 SV40 – simian virus 40
 between 1955 and 1963 millions of people worldwide received SV40-
contaminated lots of live and killed poliovirus vaccines that had been grown in
monkey cells unknowingly infected with SV40

 SV40 is detected in humans today, including in individuals too young to have

been exposed via vaccination
 may be transmitted (and other polyomaviruses) by the fecal–oral route in humans

 The prevalence of SV40 infections in humans appears to be low

 SV40 DNA has been detected in selected types of human tumors - brain tumors,
mesotheliomas, bone tumors, and lymphomas

 The role SV40 in human cancers - under investigation

52
 DNA Tumor Viruses
Polyomavirus
Pathogenesis & Pathology VII

 host range for polyomaviruses - often highly restricted

 Usually a single species can be infected and only certain cell types within that
species

 Exceptions - simian SV40 and human BK virus:

 SV40 can infect also humans and human cells

 BK virus can infect some monkeys and monkey cells

 Cell types that fail to support polyomavirus replication may be transformed by a

virus

53
 DNA Tumor Viruses
Papillomaviruses
 Family – Papillomaviridae*, 16 genera, five contain members that infect humans
(Alpha-, Beta-, Gamma-, Mupa-, and Nupapapillomavirus).
 Small, icosahedral, 55 nm in diameter, nonenveloped
 circular dsDNA, 8 kbp, cellular histones condense DNA in virion
 Replication in nucleus

 Almost 200 HPV types have been recovered - classified using molecular criteria
 Stimulate cell DNA synthesis
 Viral oncoproteins interact with cellular tumor suppressor proteins
 Significant cause of human cancer, especially cervical cancer

Two structural (capsid)

proteins - L1, L2

54

*Formerly classified in Papovaviridae family

 DNA Tumor Viruses
Papillomaviruses
 Map of the human papillomavirus genome (HPV-6, 7902 base pairs)
 The genome is circular but is shown linearized in the upstream regulatory region
(URR). The URR contains the origin of replication and enhancer and promoter
sequences.
 Early (E1–E7) and late (L1, L2) open reading frames and their functions are
shown. All the open reading frames are on the same strand of viral DNA.

In cancer cells, viral DNA is integrated into

the cellular DNA, and the E6 and E7 proteins Virus “types” are at least 10% dissimilar
are produced - interact with cellular tumor in the sequence of their L1 genes 55
suppressor proteins of p53 and Rb genes
 DNA Tumor Viruses
Papillomavirus Replication

 highly tropic for epithelial cells of the skin and mucous membranes

 Viral nucleic acid can be found in basal stem cells

 but late gene expression (capsid proteins) is restricted to the uppermost layer
of differentiated keratinocytes

 Stages in the viral replicative cycle are dependent on specific factors

that are present in sequential differentiated states of epithelial cells

 This strong dependence of viral replication on the differentiated state of

the host cell is responsible for the difficulties in propagating
papillomaviruses in vitro

56
 DNA Tumor Viruses
Papillomavirus Replication
Schematic representation of a skin wart (papilloma)

Fig. 43-9

The papillomavirus life cycle is tied to epithelial cell differentiation.

The terminal differentiation pathway of epidermal cells is shown on the left.
Events in the virus life cycle are noted on the right.
Late events in viral replication (capsid protein synthesis and virion morphogenesis)
occur only in terminally differentiated cells. 57
 DNA Tumor Viruses microabrasions allow
Papillomavirus infection of proliferating
basal layer cells at
Pathogenesis & Pathology I
other sites or within
different hosts
 transmission by close contact
 virus released from the surface of papillomatous lesions
 Papillomaviruses cause infections at cutaneous and mucosal sites

 agents of
 warts – skin warts, plantar warts, flat warts; anogenital warts, laryngeal
papillomas
 cancers - cervix, vulva, penis and anus, and a subset of head and neck cancers

 HPV genital infections - the most common STD in the US

 Cervical cancer
 the 2nd most frequent cancer in women worldwide
 a major cause of cancer deaths in developing countries
 HPV types 16 and 18 are considered to be high cancer risk
 about 16 other less common types are also considered high risk
 Many HPV types are considered benign
58
 DNA Tumor Viruses
Papillomavirus
Pathogenesis & Pathology II

 integrated copies of viral DNA - in cervical cancer cells

 in an episomal state (not integrated) in:
 noncancerous cells or premalignant lesions
 skin carcinomas
 Viral early proteins E6 and E7 are synthesized in cancer tissue
 These are HPV-transforming proteins, able to complex with Rb and p53 and other
cellular proteins
 Cell-mediated immunity is important -
 the majority of HPV infections are cleared and become undetectable within 2–3
years
 Development of HPV-associated carcinomas requires persistent infection
 Cervical cancer develops slowly, taking years to decades 59
 Types of warts
Warts can surface in groups or single spot with variety of
shapes, colors and sizes.

Most HPV viruses go away on their own depending on how strong the immune
system is. The chance of getting re-infected from the same type of HPV virus is
highly unlikely, but individual can still be infected from others
 Common skin warts Benign

HPVs - 2, 4, 27, 57

61
 plantar warts Benign

HPV 1

Plantar warts are benign and

don’t lead to cancer

Plantar wart removal

Plantar wart removal

 tend to grow
rather quickly skin warts Benign

filiform wart flat warts

flat warts

Flat warts, also called verruca

plana ,and are often found on the
face and the hands. They are
usually arranged linearly
 anogenital warts or venereal warts or
condylomata acuminata

around the anus genital warts on a female

Approximately 90% of condylomata

acuminata are related to low risk HPV
types 6 and 11, which are also
responsible for laryngeal papillomas;
dysplasias and intraepithelial
neoplasias (mucosal sites)
genital warts on a male
 In children Laryngeal papilloma, or
recurrent respiratory papillomatosis The infection is
acquired during
passage
through the birth
canal of a mother
with genital warts

~3000 cases of this

disease are diagnosed
annually - up to 3% of
children will die
 Rare medical condition
 2 per 100,000 adults and 4.5 per
100,000 children
 caused by HPV types 6 and 11
 can recur frequently, may require
repetitive surgery, and may interfere
with breathing - obstruct the larynx
 treated with surgery and antivirals
 antiangiogenic therapy shows promising 65
results
 Hand warts of butchers
 no evidence of person-to-
person transmission

 HPV-7 is distributed
throughout the population
 but only causes disease
under certain
environmental conditions
or stimuli
 and that there is some
component of animal flesh
that predisposes
activation, infection and/or
replication of HPV-7 in the
epithelia

 Bulging, cauliflower-like lesions 66

 Epidermodysplasia
verruciformis (EV)
also known as treeman syndrome
 extremely rare autosomal recessive
hereditary skin disorder
 mostly benign, but some progress to
malignancy
 characterized by abnormal
susceptibility to HPVs of the skin

 The resulting uncontrolled HPV

infections result in the growth of
scaly macules and papules,
particularly on the hands and feet
 typically associated with HPV types
5 and 8, which are found in about
80% of the normal population as
asymptomatic infections early diagnosis, and excision of the tumoral
 No curative treatment lesions take preference to prevent the
67
development of cutaneous tumors
Epidermodysplasia verruciformis (EV)
also known as treeman syndrome

68
 DNA Tumor Viruses
Papillomavirus
Clinical Findings and Epidemiology I

 HPV genital infections - the most common viral infection of the reproductive tract -
660 million people worldwide

 The peak incidence of HPV infections occurs in adolescents and young adults
under 25 years of age

 HPV-16 and HPV-18 are responsible for more than 70% of all cervical
cancers, with type 16 being most common

 HeLa cells, a widely used tissue culture cell line derived many years ago from a
cervical carcinoma, contain HPV-18 DNA

 Anal cancer is associated with high-risk HPV infection - especially at risk:

 immunocompromised patients
 men who have sex with men
69
 DNA Tumor Viruses
Papillomavirus
Clinical Findings and Epidemiology II

 Oropharyngeal cancers - a subset of head and neck squamous cell carcinomas -

are linked to HPV infections, especially by type 16:
 ~ 25% of mouth and 35% of throat cancers

 The role of men as carriers of HPV as well as vectors for transmission of infections
is well documented
 however, most penile HPV infections in men are subclinical and do not result in
HPV-associated disease

 A great multiplicity of HPV types are detected in normal skin

 asymptomatic HPV infections are acquired early in infancy
 Transmission is thought to occur from those in close contact with the child, with a
high concordance (about 60%) between types detected in infants and their mothers

 All HPV-associated cancers occur more frequently in persons with HIV/AIDS

70
 DNA Tumor Viruses
Papillomavirus
Prevention & Control-I

 Papanicolaou (Pap) smear for cervical cancer detection

 detects precancerous cellular changes in morphology, allowing for lesion
removal prior to cancer development

 HPV screening by
 DNA hybridization
 PCR methods

 Studies have indicated that primary HPV screening algorithms may be the
preferred method in most settings

 Women <20 years should not be tested - due to frequent HPV clearance in
initial infections
71
 DNA Tumor Viruses
Papillomavirus
Prevention & Control-II
 quadrivalent HPV vaccine approved in the US - in 2006
 bivalent vaccine - in 2007

 Both - noninfectious recombinant vaccines containing virus-like particles

composed of HPV L1 proteins

 quadrivalent vaccine - contains particles from HPV types 6, 11, 16, and 18
 bivalent vaccine - contains particles from types 16 and 18

 They are not effective against established HPV disease

 quadrivalent vaccine
 target population for vaccination - adolescent and young adults (males and
females)
 for prevention of cervical cancer, genital warts, and vulvar and vaginal
precancerous lesions
 vaccine-induced immunity lasts at least 10 years 72
 DNA Tumor Viruses
Adenovirus
 large group of agents widely distributed in nature
 medium-sized, nonenveloped
 linear genome of dsDNA (26–45 kbp)
 replication - species-specific, occurring in cells of the natural hosts
 commonly infect humans - mild acute illnesses - mainly of the respiratory and
intestinal tracts
 No association of adenoviruses with human neoplasms has been found

 important models for studying the molecular mechanisms by which DNA tumor
viruses usurp cellular growth control processes

 can transform rodent cells and induce the synthesis of virus-specific early
antigens that localize in both the nucleus and the cytoplasm of transformed cells
 E1A early proteins complex with the cellular Rb protein as well as with several
other cellular proteins
 early proteins, E1B and E4ORF1, bind p53 and other cellular signaling proteins
73
 DNA TUMOR VIRUSES
HEPATITIS B VIRUS
 member of the Hepadnaviridae family
 42-nm enveloped spherical virions
 circular genome of double-stranded DNA,
3.2 kbp
 one strand - incomplete and variable in
length
 studies of the virus are hampered - it has
not been grown in cell culture
 Persistent infection - an important cause of
chronic liver disease and the development
of hepatocellular carcinoma
 integration of HBV DNA may cause
insertional mutagenesis, resulting in the
activation of a cellular oncogene
 infections in adults are usually resolved these persistent hepatitis B virus
 primary infections in neonates and young infections established early in life that
children tend to become chronic in up to carry the highest risk of
90% of cases hepatocellular carcinoma later in 74life
 DNA TUMOR VIRUSES
HEPATITIS B VIRUS
 Persistent viral infection leads to necrosis, inflammation, and liver regeneration →
over time, results in cirrhosis → hepatocellular carcinoma inhibits the p53 tumor
suppressor protein
 hepatitis B virus transactivator protein - X protein - a potential viral oncoprotein
 A dietary carcinogen, aflatoxin, may be a cofactor for hepatocellular carcinoma,
especially in Africa and China
Aflatoxins - poisonous carcinogens that are produced
by certain molds (Aspergillus flavus) which grow in soil,
Peanut butter decaying vegetation, hay, and grains
 Prevention of primary infection - hepatitis B vaccine
 particularly in areas of the world where infection with hepatitis B virus is
hyperendemic - Africa, China, Southeast Asia

 woodchucks - an excellent model for hepatitis B virus infections of humans

 a similar virus - woodchuck hepatitis virus - establishes chronic infections in both
newborn and adult woodchucks - many develop hepatocellular carcinomas within a
3-year period
 currently over 350 million people worldwide are persistently infected with hepatitis B virus
leading to over 500,000 deaths from hepatocellular cancer annually 75
 DNA Tumor Viruses
Herpesviruses-1

 large viruses, diameter 125–200 nm

 linear genome of dsDNA, 125–240 kbp
 Icosahedral, enveloped
 typically cause acute infections followed by latency and eventual recurrence in
each host, including humans

 In humans, herpesviruses have been linked to several specific types of tumors

 These tumors usually contain EBV viral DNA (both integrated and episomal
forms) and viral antigens:
 Burkitt lymphoma, a tumor most commonly found in children in central Africa
 Nasopharyngeal carcinoma, more common in Cantonese Chinese and Alaskan
Eskimos
 Posttransplant lymphoproliferative disorders in immunocompromised hosts
 Hodgkin disease
76
 DNA Tumor Viruses
Herpesviruses-2
 Latent phase antigens –
 EBNAs (EBV nuclear antigen) and the LMPs (latent membrane proteins)
 synthesized by latently infected cells
 their expression reveals that an EBV genome is present
 Only EBNA1 - needed to maintain the viral DNA episomes - is invariably
expressed
 LMP1 mimics an activated growth factor receptor - viral oncogene protein
 LMP1
 cellular target - TRAF, tumor necrosis factor receptor–associated factor
 able to transform rodent fibroblasts
 essential for transformation of B lymphocytes

 EBNAs - necessary for immortalization of B cells

 EBNA1 is the only viral protein consistently expressed in Burkitt lymphoma cells

 EBV is very successful at avoiding immune elimination - may be due in part to

the function of EBNA1 - inhibition of antigen processing - infected cells escape77
killing by cytotoxic T lymphocytes
 DNA Tumor Viruses
Herpesviruses-3

 Malaria - may be a cofactor for African Burkitt lymphoma

 Most of those tumors also show characteristic chromosomal translocations

between the c-myc gene and immunoglobulin loci - result in deregulation -
activation of expression of the c-myc proto-oncogene

The c-myc protein is a transcriptional regulator that enhances

the synthesis of kinases that activate the cell cycle

 salted or dried fish - may be a dietary cofactor in EBV-related

nasopharyngeal carcinoma

 EBV-associated posttransplant lymphoproliferative disorders

 more common in highly immunosuppressed transplant patients
 can be detected earlier by screening for EBV in the blood

78
 DNA Tumor Viruses
Herpesviruses-4

 Kaposi sarcoma–associated herpesvirus - human herpesvirus 8 (KSHV/HHV8)

 is not as ubiquitous as most other human herpesviruses
 the cause of Kaposi sarcoma, primary effusion lymphoma, and multicentric
Castleman disease, a lymphoproliferative disorder
 has a number of genes related to cellular regulatory genes that may stimulate
cellular proliferation and modify host defense mechanisms

 Some herpesviruses are associated with tumors in lower animals:

 Marek disease - highly contagious lymphoproliferative disease of chickens that

can be prevented by vaccination with an attenuated strain of Marek disease virus

 The simian viruses cause inapparent infections in their natural hosts but induce
malignant T-cell lymphomas when transmitted to other species of monkeys
79
 DNA Tumor Viruses
Poxviruses
Molluscum contagiosum is a very
common skin disease that causes
red lumps or lesions to appear on
the skin
 large, brick-shaped viruses
 with a linear genome of dsDNA (130–375 kbp).

 Yaba virus produces benign tumors

(histiocytomas) in its natural host, monkeys

 Shope fibroma virus produces fibromas in some

rabbits and is able to alter cells in culture

 Molluscum contagiosum virus produces small

benign growths in humans

 Very little is known about the nature of these

proliferative diseases
80
 Tumor Viruses
3 mechanisms of tumor formation
Simple or complex
genome
RNA viruses RNA viruses DNA viruses
(mostly defective)

Virus carries onc gene Virus enhances Virus inactivates cellular

cellular oncogenes tumor suppressors genes
Directly-acting Indirectly-acting

Not in humans Human T-lymphotrophic Human papilloma virus, HPV

Virus, HTLV

Ab-MLV, Abelson murine leukemia

virus (abl oncogene)
(Gammaretrovirus)
Ab-MLV - defective
RSV, Rous sarcoma virus (src
oncogene) (Alpharetrovirus)

RSV - nondefective 81