The E6 gene of HPV 16 is sufficient for colony formation in primary human keratinocytes

1995 ◽  
Vol 121 (S1) ◽  
pp. S24-S24
Author(s):  
L. Sherman ◽  
R. Schlegel
Keyword(s):  
Colony Formation ◽  
Human Keratinocytes ◽  
Primary Human Keratinocytes ◽  
Hpv 16 ◽  
E6 Gene

scholarly journals Centrosome Abnormalities and Genomic Instability by Episomal Expression of Human Papillomavirus Type 16 in Raft Cultures of Human Keratinocytes

2001 ◽  
Vol 75 (16) ◽  
pp. 7712-7716 ◽  
Author(s):  
Stefan Duensing ◽  
Anette Duensing ◽  
Elsa R. Flores ◽  
Anh Do ◽  
Paul F. Lambert ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Genomic Instability ◽  
Transcriptional Control ◽  
Ectopic Expression ◽  
Human Keratinocytes ◽  
Primary Human Keratinocytes ◽  
Hpv 16 ◽  
Hpv E6 ◽  
Copy Numbers ◽  
E6 And E7

ABSTRACT Primary human keratinocytes with ectopic expression of high-risk human papillomavirus (HPV) E6 and E7 oncoproteins display abnormal centrosome numbers, multipolar mitoses, and aneusomy. However, it has not been explored whether these abnormalities can occur in cells containing HPV episomes where E6 and E7 expression is under viral transcriptional control. Here, we demonstrate that centrosome abnormalities and genomic instability occur in organotypic raft cultures of human keratinocytes with episomal HPV-16 even at low copy numbers. We conclude that HPV-16 DNA, when maintained as an episome, can disturb centrosome homeostasis and subvert genomic integrity of the host cell during early stages of the viral infection.

scholarly journals Human Papillomavirus (HPV) Type 18 Induces Extended Growth in Primary Human Cervical, Tonsillar, or Foreskin Keratinocytes More Effectively than Other High-Risk Mucosal HPVs

2009 ◽  
Vol 83 (22) ◽  
pp. 11784-11794 ◽  
Author(s):  
Michael J. Lace ◽  
James R. Anson ◽  
Aloysius J. Klingelhutz ◽  
John H. Lee ◽  
Aaron D. Bossler ◽  
...  
Keyword(s):  
High Risk ◽  
Human Keratinocytes ◽  
Human Papillomaviruses ◽  
Population Doubling ◽  
Primary Human Keratinocytes ◽  
Hpv 16 ◽  
Epithelial Origin ◽  
Hpv Types ◽  
Population Doublings ◽  
Hpv 18

ABSTRACT Mucosal high-risk (HR) human papillomaviruses (HPVs) that cause cervical and other anogenital cancers also are found in ∼25% of head and neck carcinomas (HNCs), especially those arising in the oropharynx and the tonsils. While many HR HPV types are common in anogenital cancer, over 90% of HPV-positive HNCs harbor HPV type 16 (HPV-16). Using a quantitative colony-forming assay, we compared the ability of full-length mucosal HPV genomes, i.e., the low-risk HPV-11 and HR HPV-16, -18, and -31, to persist in and alter the growth of primary human keratinocytes from the foreskin, cervix, and tonsils. The HR HPV types led to the formation of growing keratinocyte colonies in culture independent of the site of epithelial origin. However, HPV-18 induced colony growth in all keratinocytes >4-fold more effectively than HPV-16 or HPV-31 and >20-fold more efficiently than HPV-11 or controls. HPV-11-transfected or control colonies failed to expand beyond 32 to 36 population doublings postexplantation. In contrast, individual HR HPV-transfected clones exhibited no apparent slowdown of growth or “crisis,” and many maintained HPV plasmid persistence beyond 60 population doublings. Keratinocyte clones harboring extrachromosomal HR HPV genomes had shorter population doubling times and formed dysplastic stratified epithelia in organotypic raft cultures, mirroring the pathological features of higher-grade intraepithelial lesions, yet did not exhibit chromosomal instability. We conclude that, in culture, the HR HPV type, rather than the site of epithelial origin of the cells, determines the efficacy of inducing continued growth of individual keratinocytes, with HPV-18 being the most aggressive mucosal HR HPV type tested.

scholarly journals Eight Nucleotide Substitutions Inhibit Splicing to HPV-16 3′-Splice Site SA3358 and Reduce the Efficiency by which HPV-16 Increases the Life Span of Primary Human Keratinocytes

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e72776 ◽  
Author(s):  
Xiaoze Li ◽  
Cecilia Johansson ◽  
Carlos Cardoso Palacios ◽  
Anki Mossberg ◽  
Soniya Dhanjal ◽  
...  
Keyword(s):  
Life Span ◽  
Splice Site ◽  
Human Keratinocytes ◽  
Primary Human Keratinocytes ◽  
Nucleotide Substitutions ◽  
Hpv 16

Towards gene therapy for haemophilia B using primary human keratinocytes

1993 ◽  
Vol 3 (2) ◽  
pp. 180-183 ◽  
Author(s):  
Ann J. Gerrard ◽  
David L. Hudson ◽  
George G. Brownlee ◽  
Fiona M. Watt
Keyword(s):  
Gene Therapy ◽  
Human Keratinocytes ◽  
Primary Human Keratinocytes ◽  
Haemophilia B

Expression profiling of vitamin D treated primary human keratinocytes

2007 ◽  
Vol 100 (3) ◽  
pp. 574-592 ◽  
Author(s):  
Pamela Renate Moll ◽  
Veronika Sander ◽  
Anna-Maria Frischauf ◽  
Klaus Richter
Keyword(s):  
Vitamin D ◽  
Expression Profiling ◽  
Human Keratinocytes ◽  
Primary Human Keratinocytes

scholarly journals HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Irma Colombo ◽  
Enrico Sangiovanni ◽  
Roberta Maggio ◽  
Carlo Mattozzi ◽  
Stefania Zava ◽  
...  
Keyword(s):  
Cell Density ◽  
Skin Diseases ◽  
Inflammatory Responses ◽  
Human Keratinocytes ◽  
Suitable Model ◽  
Hacat Cells ◽  
Primary Human Keratinocytes ◽  
Proinflammatory Mediators ◽  
Cytokines And Chemokines

Cultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short culture lifetime, and variations between passages. To standardize the in vitro studies on keratinocytes, we investigated the use of HaCaT cells, a long-lived, spontaneously immortalized human keratinocyte line which is able to differentiate in vitro, as a suitable model to follow the release of inflammatory and repair mediators in response to TNFα or IL-1β. Different treatment conditions (presence or absence of serum) and differentiation stimuli (increase in cell density as a function of time in culture and elevation of extracellular calcium) were considered. ELISA and Multiplex measurement technologies were used to monitor the production of cytokines and chemokines. Taken together, the results highlight that Ca2+ concentration in the medium, cell density, and presence of serum influences at different levels the release of proinflammatory mediators by HaCaT cells. Moreover, HaCaT cells maintained in low Ca2+ medium and 80% confluent are similar to normal keratinocytes in terms of cytokine production suggesting that HaCaT cells may be a useful model to investigate anti-inflammatory interventions/therapies on skin diseases.

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