Reconstruction of 'simplified' skin: control of fabrication

Asselineau, Daniel; Pruniéras, M

doi:10.1111/j.1365-2133.1984.tb15608.x

Cited by 112 publications

(71 citation statements)

References 2 publications

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“…We have previously demonstrated that the complete HPV life cycle is strictly dependent on host tissue differentiation (5,22,49,50,52,53,58). Therefore, it was of interest to determine whether the observed changes in the expression profile of the multiple tumor suppressors upon BaP treatment was specific to differentiation, conditions which in general are conducive for virion synthesis.…”

Section: Resultsmentioning

confidence: 99%

Downregulation of Cdc2/CDK1 Kinase Activity Induces the Synthesis of Noninfectious Human Papillomavirus Type 31b Virions in Organotypic Tissues Exposed to Benzo[ a ]pyrene

Alam

Bowser

Conway

et al. 2010

J Virol

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Epidemiological studies suggest that human papillomavirus (HPV)-infected women who smoke face an increased risk for developing cervical cancer. We have previously reported that exposure of HPV-positive organotypic cultures to benzo[a]pyrene (BaP), a major carcinogen in cigarette smoke, resulted in enhanced viral titers. Since BaP is known to deregulate multiple pathways of cellular proliferation, enhanced virion synthesis could result from carcinogen/host cell interaction. Here, we report that BaP-mediated upregulation of virus synthesis is correlated to an altered balance between cell cycle-specific cyclin-dependent kinase (CDK) activity profile compared with controls. Specifically, BaP treatment increased accumulation of hyperphosphorylated retinoblastoma protein (pRb) which coincided with increased cdc2/CDK1 kinase activity, but which further conflicted with the simultaneous upregulation of CDK inhibitors p16 INK4 and p27 KIP1 , which normally mediate pRb hypophosphorylation. In contrast, p21 WAF1 and p53 levels remained unchanged. Under these conditions, CDK6 and CDK2 kinase activities were decreased, whereas CDK4 kinase activity remained unchanged. The addition of purvalanol A, a specific inhibitor of CDK1 kinase, to BaP-treated cultures, resulted in the production of noninfectious HPV type 31b (HPV31b) particles. In contrast, infectivity of control virus was unaffected by purvalanol A treatment. BaP targeting of CDK1 occurred independently of HPV status, since BaP treatment also increased CDK1 activity in tissues derived from primary keratinocytes. Our data indicate that HPV31b virions synthesized in the presence of BaP were dependent on BaP-mediated alteration in CDK1 kinase activity for maintaining their infectivity.

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Section: Resultsmentioning

confidence: 99%

Downregulation of Cdc2/CDK1 Kinase Activity Induces the Synthesis of Noninfectious Human Papillomavirus Type 31b Virions in Organotypic Tissues Exposed to Benzo[ a ]pyrene

Alam

Bowser

Conway

et al. 2010

J Virol

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“…The organotypic raft culture system has been used as an in vitro model for squamous epithelial cell differentiation (2). Epithelial cells are seeded onto a collagen gel containing feeder fibroblasts and grown under submerged conditions.…”

Section: Resultsmentioning

confidence: 99%

Epstein-Barr Virus LMP2A Transforms Epithelial Cells, Inhibits Cell Differentiation, and Activates Akt

Scholle

Bendt

Raab‐Traub

2000

J Virol

262

249

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The Epstein-Barr virus LMP2A protein was expressed in a human keratinocyte cell line, HaCaT, and effects on epithelial cell growth were detected in organotypic raft cultures and in vivo in nude mice. Raft cultures derived from LMP2A-expressing cells were hyperproliferative, and epithelial differentiation was inhibited. The LMP2A-expressing HaCaT cells were able to grow anchorage independently and formed colonies in soft agar. HaCaT cells expressing LMP2A were highly tumorigenic and formed aggressive tumors in nude mice. The LMP2A tumors were poorly differentiated and highly proliferative, in contrast to occasional tumors that arose from parental HaCaT cells and vector control cells, which grew slowly and remained highly differentiated. Animals injected with LMP2A-expressing cells developed frequent metastases, which predominantly involved lymphoid organs. Involucrin, a marker of epithelial differentiation, and E-cadherin, involved in the maintenance of intercellular contact, were downregulated in LMP2A tumors. Whereas activation of the mitogen-activated protein kinase pathway was not observed, phosphatidylinositol-3-kinase (PI3-kinase)-dependent activation of the serine-threonine kinase Akt was detected in LMP2A-expressing cells and LMP2A tumors. Inhibition of this pathway blocked growth in soft agar. These data indicate that LMP2A greatly affects cell growth and differentiation pathways in epithelial cells, in part through activation of the PI3-kinase-Akt pathway. Epstein-Barr virus (EBV) is a ubiquitous herpesvirus of the familyGammaherpesviridae. Primary infection occurs in the oropharyngeal epithelium, which is permissive for virus replication. Trafficking B lymphocytes are subsequently infected by progeny virions that go on to establish a lifelong latent infection in the B-cell compartment with periodic reactivation and reinfection of the oropharyngeal epithelium, virus release, and transmission of virus via the salivary route (27). EBV is associated with a number of human malignancies, such as Burkitt's lymphoma, Hodgkin's lymphoma, posttransplant lymphoproliferative disease, and the epithelial cell malignancy nasopharyngeal carcinoma (NPC) (27,42,(44)(45)(46)56). In EBV latency, only a subset of genes is expressed. Different types of latency can be distinguished according to the latent gene products expressed. In type I latency, characteristic of Burkitt's lymphoma, only the EBV nuclear antigen 1 (EBNA1), transcripts from the BamHI-A region of the genome (10), and the EBERs, small, nonpolyadenylated RNAs with an unknown function, are expressed. In B-cell lines immortalized by EBV in vitro, all latent genes are expressed, which include EBNA1 through -6, BamHI-A transcripts, EBERs, and three latent membrane proteins, LMP1, LMP2A, and LMP2B (latency III). The pattern of gene expression is more restricted in NPC, with expression of EBNA1, EBERs, BamHI-A transcripts, and the latent membrane proteins (latency II) (27).LMP2A and -2B arise from two different promoters, and transcription proceeds across the fused ...

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“…A cultured version of a 'skin equivalent' has been achieved through culturing of keratinocytes either on de-epidermized dermis (Regnier et al, 1981;Watt, 1988;Fartasch and Ponec, 1994) or on collagen gels embedded with dermal fibroblasts (Bell et al, 1981;Regnier et al, 1981;Asselineau and Prunieras, 1984;Asselineau et al, 1985;McCance et al, 1988;Watt, 1988;Coulomb et al, 1989;Fartasch and Ponec, 1994). Such cocultures give rise to stratified epithelium that displays many of the morphological and functional features of an epidermis in vivo (Bell et al, 1981;Kopan et al, 1987;Watt, 1988;Kopan and Fuchs, 1989;Hertle et al, 1991;Parenteau et al, 1991;Fusenig, 1994;Smola et al, 1998).…”

Section: Modeling Organs In Vitro: Organotypic Co-culturesmentioning

confidence: 99%

Modeling tissue-specific signaling and organ function in three dimensions

Schmeichel

Bissell

2003

Journal of Cell Science

537

418

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IntroductionQualitative evaluation of the cellular complexity and structural integrity of organ biopsies has been used by pathologists for decades to gain insight into human diseases. The well-accepted correlation between tissue structure and health or disease conveys important lessons for the development of experimental models for the study of normal human biology and associated disease progression. Optimally, model design should recapitulate both the 3D organization and the differentiated function of any given organ but at the same time allow experimental intervention. By doing so, cell-based models facilitate systematic analyses that address, at the molecular level, how normal organ structure and function are maintained or how the balance is lost in cancer.Because of the ethical, technical and financial constraints inherent in research on human cells and tissues, the demand for models that faithfully parallel human form and function considerably outweighs the supply. We and others asserted more than two decades ago that development of physiologically relevant models of both rodent and human origin should recognize that organs and tissues function in a 3D environment (Elsdale and Bard, 1972;Hay and Dodson, 1973;Bissell, 1981;Ingber and Folkman, 1989). Further, that in the final analysis, the organ itself is the unit of function (Bissell and Hall, 1987). We now know that exposure of cells to the spatial constraints imposed by a 3D milieu determines how cells perceive and interpret biochemical cues from the surrounding microenvironment [e.g. the extracellular matrix, growth factors and neighboring cells (for reviews, see Roskelley et al., 1995;Bissell et al., 2002;Cunha et al., 2002;Ingber, 2002;Radisky et al., 2002)]. Furthermore, it is in this biophysical and biochemical context that cells display bona fide tissue and organ specificity.Here, we describe studies of epithelial-cell-based systems that demonstrate the importance of developing and utilizing 3D human organotypic models to understand the molecular and cellular signaling events underlying human organ biology (Fig. 1). In their most simplistic form, these models comprise homogeneous epithelial cell populations that are cultured within 3D basement-membrane-like matrices. These relatively simple 'monotypic' cell models have progressively evolved into 3D co-culture models containing multiple cell types, which approximate organ structure and function in vitro and enable systematic analyses of the molecular contributions of multiple cell types. Finally, we go on to explore how human 3D culture models are being coupled to existing technologies in the mouse to generate models in vivo that could elucidate the fundamental influence of stromal-epithelial interactions in normal organ function as well as those that perturb organ homeostasis and lead to disease. As a result of these advancements, we are equipped with a hierarchy of related models that appreciate the importance of 3D environments but vary with respect to cellular complexity. By using this collectio...

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Reconstruction of 'simplified' skin: control of fabrication

Abstract: Reconstruction of' simplified' skin 221 FIGURE 3. Vertical frozen sections immuno-labelled with bullous pemphigoid antibodies. Intracellular (left) and exiracellular (righi) basemem membrane-like staining patterns in the same culture after 10 days.

Cited by 112 publications

References 2 publications

Downregulation of Cdc2/CDK1 Kinase Activity Induces the Synthesis of Noninfectious Human Papillomavirus Type 31b Virions in Organotypic Tissues Exposed to Benzo[ a ]pyrene

Downregulation of Cdc2/CDK1 Kinase Activity Induces the Synthesis of Noninfectious Human Papillomavirus Type 31b Virions in Organotypic Tissues Exposed to Benzo[ a ]pyrene

Epstein-Barr Virus LMP2A Transforms Epithelial Cells, Inhibits Cell Differentiation, and Activates Akt

Modeling tissue-specific signaling and organ function in three dimensions

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