title: E-cadherin-mediated adhesion involving Langerhans cell-like dendritic cells expanded from murine fetal skin.
abstract: Langerhans cells (LC), the epidermal contingent of the dendritic cell (DC) lineage, migrate from skin to regional lymph nodes to initiate primary immune responses against Ag encountered in skin. Because E-cadherin mediates LC-keratinocyte adhesion, E-cadherin expression and/or function must be modulated during LC migration. To facilitate studies of LC/DC cadherin biology, we defined culture conditions that allowed expansion of LC-like cells from fetal murine skin. Fetal skin-derived dendritic cells (FSDDC) were propagated from C57BL/6 day 16 fetal skin in GM-CSF- and CSF-1-supplemented media. After 14 days, aggregates of E-cadherin+ FSDDC (FSDDC-A) that resembled freshly-obtained LC with regard to phenotype and function were isolated. Nonadherent FSDDC (FSDDC-NA) with dendritic morphology, surface phenotype identical to that of interdigitating DC and potent allostimulatory capacity were released from FSDDC-A with continued incubation. A survey of cytokine mRNAs expressed by FSDDC revealed that FSDDC-A expressed predominantly TNF-alpha, TGF-beta1, and MIF mRNA. In contrast, FSDDC-NA exhibited de novo expression of IL-1beta, IL-12 (p40), increased levels of TNF-alpha and decreased MIF mRNA. Neutralizing anti-E-cadherin mAb dissociated FSDDC-A into single cells, whereas functionally inactive anti-E-cadherin mAb and mAb reactive with other adhesion molecules did not, demonstrating that adhesion within FSDDC-A was E-cadherin-mediated. FSDDC-A also preferentially adhered to E-cadherin-transfected fibroblasts. Spontaneous dissociation of FSDDC-A was accompanied by a reduction in cell surface E-cadherin expression. The availability of large numbers of cells with characteristics of LC in situ that spontaneously mature into interdigitating DC will permit detailed studies of LC/DC cadherin biology and LC/DC differentiation.
authors.affiliation: Dermatology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: T
authors.initials: T
authors.lastname: Jakob
authors.forename: A
authors.initials: A
authors.lastname: Saitoh
authors.forename: M C
authors.initials: MC
authors.lastname: Udey
mesh_terms: D000818:Animals; D015820:Cadherins / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D016207:Cytokines / Q000502:physiology; D003713:Dendritic Cells / Q000166:cytology* / Q000502:physiology; D005260:Female; D005434:Flow Cytometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D012867:Skin / Q000166:cytology* / Q000196:embryology
chemical_list: D015820:Cadherins; D016207:Cytokines