Organization of Human Papillomavirus Productive Cycle during Neoplastic Progression Provides a Basis for Selection of Diagnostic Markers

ABSTRACT The productive cycle of human papillomaviruses (HPVs) can be divided into discrete phases. Cell proliferation and episomal maintenance in the lower epithelial layers are followed by genome amplification and the expression of capsid proteins. These events, which occur in all productive infections, can be distinguished by using antibodies to viral gene products or to surrogate markers of their expression. Here we have compared precancerous lesions caused by HPV type 16 (HPV16) with lesions caused by HPV types that are not generally associated with human cancer. These include HPV2 and HPV11, which are related to HPV16 (supergroup A), as well as HPV1 and HPV65, which are evolutionarily divergent (supergroups E and B). HPV16-induced low-grade squamous intraepithelial lesions (CIN1) are productive infections which resemble those caused by other HPV types. During progression to cancer, however, the activation of late events is delayed, and the thickness of the proliferative compartment is progressively increased. In many HPV16-induced high-grade squamous intraepithelial lesions (CIN3), late events are restricted to small areas close to the epithelial surface. Such heterogeneity in the organization of the productive cycle was seen only in lesions caused by HPV16 and was not apparent when lesions caused by other HPV types were compared. By contrast, the order in which events in the productive cycle were initiated was invariant and did not depend on the infecting HPV type or the severity of disease. The distribution of viral gene products in the infected cervix depends on the extent to which the virus can complete its productive cycle, which in turn reflects the severity of cervical neoplasia. It appears from our work that the presence of such proteins in cells at the epithelial surface allows the severity of the underlying disease to be predicted and that markers of viral gene expression may improve cervical screening.

[1]  E. Villiers.  Taxonomic classification of papillomaviruses , 2001 .

[2]  M. Ozbun,et al.  Human papillomavirus type 31b E1 and E2 transcript expression correlates with vegetative viral genome amplification. , 1998, Virology.

[3]  H. Selinka,et al.  Analysis of the infectious entry pathway of human papillomavirus type 33 pseudovirions. , 2002, Virology.

[4]  I. Frazer,et al.  Identification of the alpha6 integrin as a candidate receptor for papillomaviruses , 1997, Journal of Virology.

[5]  S. Syrjänen,et al.  Integrated Human Papillomavirus Type 16 Is Frequently Found in Cervical Cancer Precursors as Demonstrated by a Novel Quantitative Real-Time PCR Technique , 2002, Journal of Clinical Microbiology.

[6]  G. Williams,et al.  Minichromosome maintenance proteins as biological markers of dysplasia and malignancy. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[7]  M. Stanley,et al.  The interaction between human papillomavirus type 16 E1 and E2 proteins is blocked by an antibody to the N-terminal region of E2. , 1995, European journal of biochemistry.

[8]  J. Millar,et al.  Identification of a G2 Arrest Domain in the E1∧E4 Protein of Human Papillomavirus Type 16 , 2002, Journal of Virology.

[9]  C. Crum,et al.  Viral and histopathologic correlates of MN and MIB-1 expression in cervical intraepithelial neoplasia. , 1996, Human pathology.

[10]  M. Ozbun,et al.  Characterization of late gene transcripts expressed during vegetative replication of human papillomavirus type 31b , 1997, Journal of virology.

[11]  H. Fox,et al.  "CERVICAL INTRAEPITHELIAL NEOPLASIA" , 1982, The Lancet.

[12]  M. von Knebel Doeberitz,et al.  Overexpression of p16INK4A as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri , 2001, International journal of cancer.

[13]  R Reid,et al.  Human papillomavirus infection of the cervix: relative risk associations of 15 common anogenital types. , 1992, Obstetrics and gynecology.

[14]  A. Venuti,et al.  Egg yolk antibodies against the E7 oncogenic protein of human papillomavirus type 16 , 2001, Archives of Virology.

[15]  T. R. Broker,et al.  Induction of proliferating cell nuclear antigen in differentiated keratinocytes of human papillomavirus-infected lesions. , 1994, Human pathology.

[16]  G. Williams,et al.  Improved cervical smear assessment using antibodies against proteins that regulate DNA replication. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Lambert,et al.  Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells , 1995, Journal of virology.

[18]  C. Crum,et al.  Coexpression of the human papillomavirus type 16 E4 and L1 open reading frames in early cervical neoplasia. , 1990, Virology.

[19]  J. Doorbar,et al.  Sequence divergence yet conserved physical characteristics among the E4 proteins of cutaneous human papillomaviruses. , 1989, Virology.

[20]  H. B. Lim,et al.  Human papillomavirus type 31b late gene expression is regulated through protein kinase C-mediated changes in RNA processing , 1995, Journal of virology.

[21]  C. Meijer,et al.  Penile lesions and human papillomavirus in male sexual partners of women with cervical intraepithelial neoplasia. , 2002, Journal of the American Academy of Dermatology.

[22]  A. Ishimoto,et al.  Modulation of the Cell Division Cycle by Human Papillomavirus Type 18 E4 , 2002, Journal of Virology.

[23]  G. Snow,et al.  The use of general primers in the polymerase chain reaction permits the detection of a broad spectrum of human papillomavirus genotypes. , 1990, The Journal of general virology.

[24]  H. Moch,et al.  Association of p27Kip1, cyclin E and c-myc expression with progression and prognosis in HPV-positive cervical neoplasms. , 1998, Anticancer research.

[25]  P. Lambert,et al.  Integration of human papillomavirus type 16 DNA into the human genome leads to increased stability of E6 and E7 mRNAs: implications for cervical carcinogenesis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[26]  A. Giannoudis,et al.  Variation in the E2-binding domain of HPV 16 is associated with high-grade squamous intraepithelial lesions of the cervix , 2001, British Journal of Cancer.

[27]  D. Lowy,et al.  Positive and negative regulation of cell proliferation by E2F-1: influence of protein level and human papillomavirus oncoproteins , 1994, Molecular and cellular biology.

[28]  J. Doorbar,et al.  Identification of the human papilloma virus‐1a E4 gene products. , 1986, The EMBO journal.

[29]  S. Bingham,et al.  Analysis of minichromosome maintenance proteins as a novel method for detection of colorectal cancer in stool , 2002, The Lancet.

[30]  K. Jansen,et al.  The L1 Major Capsid Protein of Human Papillomavirus Type 11 Recombinant Virus-like Particles Interacts with Heparin and Cell-surface Glycosaminoglycans on Human Keratinocytes* , 1999, The Journal of Biological Chemistry.

[31]  F. X. Bosch,et al.  Epidemiologic classification of human papillomavirus types associated with cervical cancer. , 2003, The New England journal of medicine.

[32]  J. Doorbar,et al.  Identification of proteins encoded by the L1 and L2 open reading frames of human papillomavirus 1a , 1987, Journal of virology.

[33]  Young Tae Kim,et al.  Expression of cyclin E and p27(KIP1) in cervical carcinoma , 2000 .

[34]  K. Cooper,et al.  Biotinyl-Tyramide-Based In Situ Hybridization Signal Patterns Distinguish Human Papillomavirus Type and Grade of Cervical Intraepithelial Neoplasia , 2002, Modern Pathology.

[35]  F. Talamantes,et al.  Sensitivity of the cervical transformation zone to estrogen-induced squamous carcinogenesis. , 2000, Cancer research.

[36]  J. Nevins,et al.  Cellular targets for activation by the E2F1 transcription factor include DNA synthesis- and G1/S-regulatory genes , 1995, Molecular and cellular biology.

[37]  S. J. Kim,et al.  Regulation of cell growth and HPV genes by exogenous estrogen in cervical cancer cells. , 2000, International journal of gynecological cancer : official journal of the International Gynecological Cancer Society.

[38]  M. Yamaguchi,et al.  Essential Role of E2F Recognition Sites in Regulation of the Proliferating Cell Nuclear Antigen Gene Promoter during Drosophila Development (*) , 1995, The Journal of Biological Chemistry.

[39]  K. Sotlar,et al.  Life Cycle Heterogeneity in Animal Models of Human Papillomavirus-Associated Disease , 2002, Journal of Virology.

[40]  Daisy R. Lee,et al.  Association of human papillomavirus type 16 integration in the E2 gene with poor disease‐free survival from cervical cancer , 1997 .

[41]  M. Campion,et al.  Analysis of the physical state of different human papillomavirus DNAs in intraepithelial and invasive cervical neoplasm , 1991, Journal of virology.

[42]  A. Teyssié,et al.  Physical status of the E2 human papilloma virus 16 viral gene in cervical preneoplastic and neoplastic lesions. , 2001, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[43]  T. R. Broker,et al.  Identification and mapping of human papillomavirus type 1 RNA transcripts recovered from plantar warts and infected epithelial cell cultures , 1987, Journal of virology.

[44]  T. Iftner,et al.  Identification of a differentiation-inducible promoter in the E7 open reading frame of human papillomavirus type 16 (HPV-16) in raft cultures of a new cell line containing high copy numbers of episomal HPV-16 DNA , 1996, Journal of virology.

[45]  P. Autier,et al.  Transformation zone location and intraepithelial neoplasia of the cervix uteri. , 1996, British Journal of Cancer.

[46]  E. Flores,et al.  Establishment of the human papillomavirus type 16 (HPV-16) life cycle in an immortalized human foreskin keratinocyte cell line. , 1999, Virology.

[47]  M. Rubin,et al.  Detection and typing of human papillomavirus DNA in penile carcinoma: evidence for multiple independent pathways of penile carcinogenesis. , 2001, The American journal of pathology.

[48]  W. Chien,et al.  p21cip1 Degradation in Differentiated Keratinocytes Is Abrogated by Costabilization with Cyclin E Induced by Human Papillomavirus E7 , 2001, Journal of Virology.

[49]  Kiyoshi Ohtani,et al.  Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F , 1999, Oncogene.

[50]  A. Kaufmann,et al.  Regulation of human papillomavirus type 18 in vivo: effects of estrogen and progesterone in transgenic mice. , 1997, Gynecologic oncology.

[51]  R. Schlegel,et al.  Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. , 1985, The American journal of pathology.

[52]  E. Flores,et al.  The Human Papillomavirus Type 16 E7 Oncogene Is Required for the Productive Stage of the Viral Life Cycle , 2000, Journal of Virology.

[53]  A. Redkar,et al.  Oestrogen increases S-phase fraction and oestrogen and progesterone receptors in human cervical cancer in vivo. , 1997, British Journal of Cancer.

[54]  L. Huang,et al.  Differential effects of progestins and estrogens on long control regions of human papillomavirus types 16 and 18. , 1996, Biochemical and biophysical research communications.

[55]  M. Stoler,et al.  p16 Immunohistochemistry Improves Interobserver Agreement in the Diagnosis of Cervical Intraepithelial Neoplasia , 2002 .

[56]  J. Peto,et al.  Human papillomavirus is a necessary cause of invasive cervical cancer worldwide , 1999, The Journal of pathology.

[57]  S. Riethdorf,et al.  Human papillomaviruses, expression of p16, and early endocervical glandular neoplasia. , 2002, Human pathology.

[58]  J. Ferlay,et al.  Globocan 2000 : cancer incidence, mortality and prevalence worldwide , 2001 .

[59]  S. Labrecque,et al.  DNA damage induced p53 mediated transcription is inhibited by human papillomavirus type 18 E6. , 1994, Oncogene.

[60]  A. W. Norhanum,et al.  Human papillomavirus DNA and virus-encoded antigens in cervical carcinoma. , 1997, The Medical journal of Malaysia.

[61]  G. Pfeifer,et al.  In Vivo Structure of Two Divergent Promoters at the Human PCNA Locus , 1999, The Journal of Biological Chemistry.

[62]  M. Wells,et al.  Inverse relationship between the expression of the human papillomavirus type 16 transcription factor E2 and virus DNA copy number during the progression of cervical intraepithelial neoplasia. , 2000, The Journal of general virology.

[63]  M. von Knebel Doeberitz,et al.  p16INK4a Immunohistochemistry Improves Interobserver Agreement in the Diagnosis of Cervical Intraepithelial Neoplasia , 2002, The American journal of surgical pathology.

[64]  B. Quade,et al.  Ki-67, Cyclin E, and p16 INK4 Are Complimentary Surrogate Biomarkers for Human Papilloma Virus-Related Cervical Neoplasia , 2001, The American journal of surgical pathology.

[65]  J. Melnick,et al.  Quantitation of papova virus particles in human warts. , 1962, Journal of the National Cancer Institute.

[66]  K. Choo,et al.  Integration of human papillomavirus type 16 into cellular DNA of cervical carcinoma: preferential deletion of the E2 gene and invariable retention of the long control region and the E6/E7 open reading frames. , 1987, Virology.

[67]  J. Brisson,et al.  Risk factors for cervical intraepithelial neoplasia: differences between low- and high-grade lesions. , 1994, American journal of epidemiology.

[68]  T. Albrecht,et al.  Persistent Viral Infections , 1996 .

[69]  Steven Wolinsky,et al.  Human papillomavirus types 6 and 11 mRNAs from genital condylomata acuminata , 1987, Journal of virology.

[70]  M. Stanley,et al.  Production and characterisation of a monoclonal antibody to human papillomavirus type 16 using recombinant vaccinia virus. , 1990, Journal of clinical pathology.

[71]  L. Nasir,et al.  Bovine papillomaviral gene expression in equine sarcoid tumours. , 1999, Virus research.

[72]  R. Laskey,et al.  Detection of S-phase cells in tissue sections by in situ DNA replication , 2000, Nature Cell Biology.

[73]  Raju Gc.  Expression of the proliferating cell nuclear antigen in cervical neoplasia. , 1994 .

[74]  R. Dulbecco.  Cell transformation by viruses. , 1969, Science.

[75]  Peter Baldwin,et al.  Translational approaches to improving cervical screening , 2003, Nature Reviews Cancer.

[76]  T. Ince,et al.  Surrogate Biomarkers of HPV Infection in Cervical Neoplasia Screening and Diagnosis , 2001, Advances in anatomic pathology.

[77]  Luise Florin,et al.  Human Papillomavirus Infection Requires Cell Surface Heparan Sulfate , 2001, Journal of Virology.

[78]  E. Androphy,et al.  Identification of human papillomavirus type 18 E6 polypeptide in cells derived from human cervical carcinomas. , 1987, The Journal of general virology.

[79]  S. Fox,et al.  Minichromosome maintenance protein 2 expression in normal kidney and renal cell carcinomas: relationship to tumor dormancy and potential clinical utility. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[80]  A. Venuti,et al.  HPV16 and HPV18 in genital tumors: Significantly different levels of viral integration and correlation to tumor invasiveness , 2002, Journal of medical virology.

[81]  S. Mitrani‐Rosenbaum,et al.  Oestrogen stimulates differential transcription of human papillomavirus type 16 in SiHa cervical carcinoma cells. , 1989, The Journal of general virology.

[82]  M. Stanley,et al.  A C-Terminal Helicase Domain of the Human Papillomavirus E1 Protein Binds E2 and the DNA Polymerase α-Primase p68 Subunit , 1998, Journal of Virology.

[83]  H. Sato,et al.  Human papillomavirus type 16 E6 proteins with glycine substitution for cysteine in the metal-binding motif. , 1991, Virology.

[84]  Shingo Suzuki,et al.  E7 Proteins of four groups of human papillomaviruses, irrespective of their tissue tropism or cancer association, possess the ability to transactivate transcriptional promoters E2F site dependently , 2004, Virus Genes.

[85]  P. Rust,et al.  Human Papilloma Virus (HPV)‐E6/E7 and Epidermal Growth Factor Receptor (EGF‐R) Protein Levels in Cervical Cancer and Cervical Intraepithelial Neoplasia (CIN) , 2001, American journal of reproductive immunology.

[86]  D. Spitkovsky,et al.  The human papillomavirus type 16 E7 protein is associated with the nucleolus in mammalian and yeast cells , 1997, Oncogene.

[87]  K. Stoeber,et al.  Diagnosis of genito-urinary tract cancer by detection of minichromosome maintenance 5 protein in urine sediments. , 2002, Journal of the National Cancer Institute.

[88]  R. Knippers,et al.  Stability of the replicative Mcm3 protein in proliferating and differentiating human cells. , 1998, Experimental cell research.

[89]  T. Nakajima,et al.  Expression status of p16 protein is associated with human papillomavirus oncogenic potential in cervical and genital lesions. , 1998, The American journal of pathology.

[90]  G. Raju.  Expression of the proliferating cell nuclear antigen in cervical neoplasia. , 1994, International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists.

[91]  S. Napthine,et al.  Characterization of events during the late stages of HPV16 infection in vivo using high-affinity synthetic Fabs to E4. , 1997, Virology.