title: IL-12 is expressed and released by human keratinocytes and epidermoid carcinoma cell lines.
abstract: IL-12 is a 70-kDa heterodimeric cytokine composed of two covalently linked chains, p40 and p35. IL-12 has multiple effects on T and NK cells and recently was found to be required for optimal Th1 cell development. In a variety of inflammatory skin disorders including delayed type hypersensitivity and contact hypersensitivity, Th1 cells appear to be critically involved. Because keratinocytes are well known to exhibit the capacity to release a variety of proinflammatory and immunologic cytokines, and thereby to be able to modulate inflammatory and immune reactions within the skin, it was investigated whether human keratinocytes and keratinocyte cell lines can release IL-12. Supernatants of phorbol-12,13-dibutyrate (PDBu) stimulated human epidermoid carcinoma cell lines (KB, A431) induced IFN-gamma production by PBL. This activity could be completely blocked by the addition of an anti-IL-12 Ab directed against the p40 subunit of IL-12 (C8.6). Release of IL-12 by keratinocyte cell lines was further confirmed by an RIA specific for p40. Immunoprecipitation under reducing conditions using the C8.6 Ab yielded specific bands at 40 kDa in supernatants of both KB cells and normal human keratinocytes. Northern blot analysis revealed IL-12-specific mRNA transcripts in PDBu-treated KB cells using cDNA probes encoding for p35 and p40. Moreover, by reverse transcription PCR specific transcripts for p35 and p40 were found in keratinocytes. These data indicate that keratinocyte cell lines and, although to a much lesser degree, normal human keratinocytes, exhibit the capacity to make IL-12; thus demonstrating for the first time IL-12 production by nonhemopoietic cells. Thus, one may speculate that keratinocytes via this capacity may influence the fate of Th-mediated immune responses, favoring Th1 responses by enhanced production of IL-12 or Th2 responses by reduced IL-12 release.
authors.affiliation: Ludwig Boltzmann Institute for Cellbiology and Immunobiology of the Skin, Department of Dermatology, University of Münster, Germany.
authors.forename: Y
authors.initials: Y
authors.lastname: Aragane
authors.forename: H
authors.initials: H
authors.lastname: Riemann
authors.forename: R S
authors.initials: RS
authors.lastname: Bhardwaj
authors.forename: A
authors.initials: A
authors.lastname: Schwarz
authors.forename: Y
authors.initials: Y
authors.lastname: Sawada
authors.forename: H
authors.initials: H
authors.lastname: Yamada
authors.forename: T A
authors.initials: TA
authors.lastname: Luger
authors.forename: M
authors.initials: M
authors.lastname: Kubin
authors.forename: G
authors.initials: G
authors.lastname: Trinchieri
authors.forename: T
authors.initials: T
authors.lastname: Schwarz
mesh_terms: D001483:Base Sequence; D001681:Biological Assay; D015152:Blotting, Northern; D002294:Carcinoma, Squamous Cell / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000378:metabolism*; D015603:Keratinocytes / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012313:RNA / Q000302:isolation & purification; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
chemical_list: D018664:Interleukin-12; D012313:RNA; D007371:Interferon-gamma