Culture of Human Cervical Epithelial Cells

Stanley, Margaret

doi:10.1002/0471221201.ch5

Cited by 12 publications

(17 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similar to normal ectocervical cells in monolayer culture (33), colonies of W12p9 comprised a heterogeneous population showing some evidence of stratification and differentiation ( Fig. 2A).…”

Section: Confirmation That Hpv16 In W12p9 Is Exclusively Episomalmentioning

confidence: 88%

Acquisition of High-Level Chromosomal Instability Is Associated with Integration of Human Papillomavirus Type 16 in Cervical Keratinocytes

et al. 2004

Self Cite

View full text Add to dashboard Cite

Whereas two key steps in cervical carcinogenesis are integration of high-risk human papillomavirus (HR-HPV) and acquisition of an unstable host genome, the temporal association between these events is poorly understood. Chromosomal instability is induced when HR-HPV E7 oncoprotein is overexpressed from heterologous promoters in vitro. However, it is not known whether such events occur at the "physiologically" elevated levels of E7 produced by deregulation of the homologous HR-HPV promoter after integration. Indeed, an alternative possibility is that integration in vivo is favored in an already unstable host genome. We have addressed these issues using the unique human papillomavirus (HPV) 16-containing cervical keratinocyte cell line W12, which was derived from a low-grade squamous intraepithelial lesion and thus acquired HPV16 by "natural" infection. Whereas W12 at low passage contains HPV16 episomes only, long-term culture results in the emergence of cells containing integrated HPV16 only. We show that integration of HPV16 in W12 is associated with 3 deletion of the E2 transcriptional repressor, resulting in deregulation of the homologous promoter of the integrant and an increase in E7 protein levels. We further demonstrate that high-level chromosomal instability develops in W12 only after integration and that the forms of instability observed correlate with the physical state of HPV16 DNA and the level of E7 protein. Whereas intermediate E7 levels are associated with numerical chromosomal abnormalities, maximal levels are associated with both numerical and structural aberrations. HR-HPV integration is likely to be a critical event in cervical carcinogenesis, preceding the development of chromosomal abnormalities that drive malignant progression.

show abstract

Section: Confirmation That Hpv16 In W12p9 Is Exclusively Episomalmentioning

confidence: 88%

Acquisition of High-Level Chromosomal Instability Is Associated with Integration of Human Papillomavirus Type 16 in Cervical Keratinocytes

et al. 2004

Self Cite

View full text Add to dashboard Cite

show abstract

“…Formation of desmosome-like structures is a marker of differentiation of cervical squamous epithelial cells [16], such as ME180 cells. These structures are characterized by the presence of typical tonofilaments rich in keratin.…”

Section: Colocalization Of Gbs With Both Tight and Adherent Junctionsmentioning

confidence: 99%

Group BStreptococcusCrosses Human Epithelial Cells by a Paracellular Route

Soriano¹,

Santi²,

Taddei³

et al. 2006

J INFECT DIS

View full text Add to dashboard Cite

Colonization of the colon and vagina is thought to be important in the pathogenesis of group B Streptococcus (GBS) infection. However, little is known about the strategies used by GBS to translocate through the epithelial barrier during the onset of disease. We used differentiated epithelial cells grown on transwell inserts as a model of the epithelial barrier. Bacterial translocation occurred without a detectable decrease in transepithelial resistance. Whereas acapsular GBS was better able to adhere to and invade epithelial cells, the percentage of bacteria translocating across the epithelial monolayer was independent of the presence of the capsule. Transmission electron microscopy showed the intimate association of GBS with intercellular junctions and the capacity to cross the monolayer by a paracellular mechanism. This process consisted of an active and transient opening of cell junctions. Indeed, GBS was preferentially found along the cell perimeter, where it colocalized with junctional protein complexes.

show abstract

“…Further investigations were carried out to exclude a low level of HIV infection of genital epithelial cells. In vitro cultures of primary ectocervical and endocervical monolayers were established based on previously described methods (37). Epithelial monolayers were exposed to either cell-free M-tropic HIV-1 BaL or T-tropic HIV-1 RF , or to cell-associated HIV in the form of PM-1 cells (a T-cell line expressing both CCR5 and CXCR4 coreceptors) or human PBMC, chronically infected with either BaL or RF.…”

Section: Primary Target Cells Of Hiv Infection Within Genital Mucosamentioning

confidence: 99%

Parameters of Human Immunodeficiency Virus Infection of Human Cervical Tissue and Inhibition by Vaginal Virucides

Greenhead

Hayes

Watts

et al. 2000

J Virol

272

262

View full text Add to dashboard Cite

Heterosexual transmission of human immunodeficiency virus (HIV) is the most frequent mode of infectionworldwide. However, the immediate events between exposure to infectious virus and establishment of infection are still poorly understood. This study investigates parameters of HIV infection of human female genital tissue in vitro using an explant culture model. In particular, we investigated the role of the epithelium and virucidal agents in protection against HIV infection. We have demonstrated that the major target cells of infection reside below the genital epithelium, and thus HIV must cross this barrier to establish infection. Immune activation enhanced HIV infection of such subepithelial cells. Furthermore, our data suggest that genital epithelial cells were not susceptible to HIV infection, appear to play no part in the transfer of infectious virus across the epithelium, and thus may provide a barrier to infection. In addition, experiments using a panel of virucidal agents demonstrated differential efficiency to block HIV infection of subepithelial cells from partial to complete inhibition. This is the first demonstration that virucidal agents designed for topical vaginal use block HIV infection of genital tissue. Such agents have major implications for world health, as they will provide women with a mechanism of personal and covert protection from HIV infection.

show abstract

Culture of Human Cervical Epithelial Cells

Cited by 12 publications

References 20 publications

Acquisition of High-Level Chromosomal Instability Is Associated with Integration of Human Papillomavirus Type 16 in Cervical Keratinocytes

Acquisition of High-Level Chromosomal Instability Is Associated with Integration of Human Papillomavirus Type 16 in Cervical Keratinocytes

Group BStreptococcusCrosses Human Epithelial Cells by a Paracellular Route

Parameters of Human Immunodeficiency Virus Infection of Human Cervical Tissue and Inhibition by Vaginal Virucides

Contact Info

Product

Resources

About