Reciprocal regulation of permeability through a cultured keratinocyte sheet by IFN-gamma and IL-4.
The T cell cytokines profoundly modify the phenotypic and functional characteristics of keratinocytes. Until now, no study has focused on the effect of Th1 and Th2 cytokines on keratinocyte permeability. Using a two-layer well culturing system, permeability was assessed through cultured keratinocyte sheet in the presence or absence of various concentrations of IFN-gamma and IL-4. Transepithelial electrical resistance (TER) and the flux of 40 kDa FITC-dextrans were measured across the cultured keratinocyte sheet. IFN-gamma significantly increased the TER in a dose- and time-dependent manner, suggesting that IFN-gamma profoundly inhibited the permeability of ions through the keratinocyte sheet. In contrast, IL-4 did not affect the TER. When compared to medium control, the flux of FITC-dextran of the IFN-gamma group was significantly decreased in a dose-dependent fashion. In sharp contrast, the flux of FITC-dextran was significantly and dose-dependently increased in the presence of IL-4. A significant increase in TER and a significant decrease in the flux of dextran suggested that IFN-gamma clearly reduced the permeability of both ions and high molecular weight material through the keratinocyte sheet. Although IL-4 did not affect the permeability of the ions, it significantly enhanced the permeability of high molecular weight material. A flow cytometric assay revealed that the expression of desmoglein-3 was suppressed by IL-4, but was enhanced by IFN-gamma. The reciprocal regulation of permeability of the cultured keratinocyte sheet by IFN-gamma and IL-4 may be partly related to the modification of intercellular adhesion molecules.
Kobayashi
Junichi
J
Department of Developmental Molecular Anatomy, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan. jkobayashi@jcom.home.ne.jp
Inai
Tetsuichiro
T
Morita
Keisuke
K
Moroi
Yoichi
Y
Urabe
Kazunori
K
Shibata
Yosaburo
Y
Furue
Masutaka
M
D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D051184:Desmoglein 3; D051182:Desmogleins; D051180:Desmoplakins; D003911:Dextrans; D015536:Down-Regulation / Q000187:drug effects; D017097:Electric Impedance; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D015847:Interleukin-4 / Q000494:pharmacology*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects
D003598:Cytoskeletal Proteins; D051184:Desmoglein 3; D051182:Desmogleins; D051180:Desmoplakins; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D015847:Interleukin-4; D007371:Interferon-gamma; D016650:Fluorescein-5-isothiocyanate