Vasoactive intestinal peptide and cytokines enhance stem cell factor production from epidermal keratinocytes DJM-1.
Stem cell factor can induce mast cell proliferation and melanocyte activation. Vasoactive intestinal peptide has been suggested to play a part in inflammatory dermatoses, such as atopic dermatitis. The aim of this study was to investigate the possible role of stem cell factor in atopic dermatitis by analyzing epidermal stem cell factor production induced by vasoactive intestinal peptide and cytokines. Full-length type stem cell factor transcript was detected in normal human epidermal keratinocytes, and a human epidermal keratinocyte cell line DJM-1, as well as normal human dermal fibroblasts, using reverse transcription-polymerase chain reaction. Spliced-type stem cell factor transcript was detected in both DJM-1 cells and normal human epidermal keratinocytes. Western blot analysis with stem cell factor antibody revealed a protein of the known molecular size of membrane-bound stem cell factor in the lysates of all three cell types. Stem cell factor immunoreactivity was found in the cytoplasm and the membrane of both DJM-1 cells and normal human epidermal keratinocytes using confocal laser scanning microscope. We examined the effects of vasoactive intestinal peptide and cytokines on stem cell factor production of DJM-1 cells using enzyme-linked immunosorbent assays. Stem cell factor contents significantly increased in culture supernatants of DJM-1 cells treated with 1000 nm vasoactive intestinal peptide and/or cytokines, including interleukins 4 and 13, tumor necrosis factor-alpha, and interferon-gamma. Overall, these results suggest that several inflammatory cytokines (T helper 1 and 2) and vasoactive intestinal peptide from mast cells and nerve endings are capable of inducing stem cell factor production from epidermal keratinocytes in atopic dermatitis.
Kakurai
Maki
M
Department of Dermatology, Center for Molecular Medicine, Jichi Medical School, School of Medicine, Tochigi, Japan. kwackie@jichi.ac.jp
Fujita
Nobuya
N
Kiyosawa
Tomoharu
T
Inoue
Tae
T
Ishibashi
Shun
S
Furukawa
Yusuke
Y
Demitsu
Toshio
T
Nakagawa
Hidemi
H
D017398:Alternative Splicing; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D016207:Cytokines / Q000494:pharmacology*; D003593:Cytoplasm / Q000737:chemistry; D003876:Dermatitis, Atopic / Q000503:physiopathology; D004797:Enzyme-Linked Immunosorbent Assay; D000078404:Epidermal Cells; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D018613:Microscopy, Confocal; D012333:RNA, Messenger / Q000032:analysis; D019089:Stem Cell Factor / Q000032:analysis / Q000235:genetics*; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology*; D014665:Vasodilator Agents / Q000494:pharmacology*
D016207:Cytokines; D012333:RNA, Messenger; D019089:Stem Cell Factor; D014665:Vasodilator Agents; D014660:Vasoactive Intestinal Peptide