Establishment of a cell line with features of early dendritic cell precursors from fetal mouse skin

Giampiero Girolomoni, Manfred B. Lutz, Saveria Pastore, Caroline U. Aßmann, Andrea Cavani, Paola Ricciardi-Castagnoli

Research output: Contribution to journalArticlepeer-review

Abstract

During ontogeny, the skin is progressively populated by major histocompatibility complex class II-negative dendritic cell (DC) precursors that then mature into efficient antigen-presenting cells (APC). To characterize these DC progenitors better, we generated myeloid cell lines from fetal mouse skin by infecting cell suspensions with a retroviral vector carrying an env(AKR)-myc(MH2) fusion gene. These cells, represented by the line FSDC, displayed a dendritic morphology and their proliferation in serum-free medium was promoted by granulocyte/macrophage colony-stimulating factor (GM-CSF), but not macrophage-CSF. FSDC expressed strong surface-membrane ATP/ADPase activity, intracellular staining for 2A1 antigen, and a surface phenotype consistent with a myeloid precursor: H-2(d,b+), I-A(d,b+), CD54+, CD11b+, CD11c+, 2.4G2+, F4/80+, CD44+, 2F8+, ERMP 12-, Sca-1+, Sca-2+, NLDC-145-, B7.2+, B7.1-, J11d-, B220-, Thy-1-, and CD3-. FSDC stimulated poorly allogeneic or syngeneic T cells in the primary mixed-leukocyte reaction, and markedly increased this function after treatment with GM-CSF, GM-CSF and interleukin (IL)-4 or interferon-γ (IFN-γ); in contrast, stem cell factor, IL-1α and tumor necrosis factor-α had no effect. Preculture with IFN-γ was required for presentation of haptens to primed T cells in vitro. However, FSDC; even after cytokine activation, were less potent APC than adult epidermal Langerhans cells in both of the above assays. Finally, FSDC derivatized with haptens and injected either intravenously or subcutaneously could efficiently induce contact sensitivity responses in naive syngeneic mice. The results indicate that fetal mouse skin is colonized by myeloid precursors possessing a macrophage/immature DC-like surface phenotype and priming capacity in vivo. These cells need further differentiation and activation signals (e.g. cytokines) to express their antigen presenting potential in vitro.

Original languageEnglish
Pages (from-to)2163-2169
Number of pages7
JournalEuropean Journal of Immunology
Volume25
Issue number8
DOIs
Publication statusPublished - Aug 1995

Keywords

  • Cell immortalization
  • Dendritic cell activation
  • Dendritic cell development

ASJC Scopus subject areas

  • Immunology

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