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Biochem J. Jan 15, 1996; 313(Pt 2): 625–631.
PMCID: PMC1216953

Granulocyte/macrophage colony-stimulating factor is an intrinsic keratinocyte-derived growth factor for human melanocytes in UVA-induced melanosis.

Abstract

Recently we demonstrated that endothelins secreted from human keratinocytes act as intrinsic mitogens and melanogens for human melanocytes in UVB-induced melanosis. We show here that UVA-induced melanosis is associated with other keratinocyte-derived growth factors, secretion of which is specifically stimulated after exposure of human keratinocytes to UVA. Medium conditioned by UVA-exposed human keratinocytes elicited a significant increase in DNA synthesis by cultured human melanocytes in a UVA dose-dependent manner. Analysis of endothelin-1 and interleukin (IL)-1 alpha in the conditioned medium by ELISA, both of which are major keratinocyte-derived cytokines involved in UVB-associated melanocyte activation, revealed that UVA exposure did not cause human keratinocytes to stimulate the secretion of the two cytokines. In contrast, the levels of several other cytokines such as IL-6, IL-8 and granulocyte/macrophage colony-stimulating factor (GM-CSF) were significantly increased in the conditioned medium of human keratinocytes after exposure to UVA at a dose of 1.0 J/cm2. The gel chromatographic profile of UVA-exposed keratinocyte-conditioned medium demonstrated that there were two factors (P-1 and P-2) with molecular masses of approx. 20 and 1 kDa respectively that stimulate DNA synthesis in human melanocytes, and the larger species (P-1) also increased melanization as assessed by [14C]thiouracil incorporation. Quantitative analysis of cytokines in chromatographic fractions by ELISA revealed the P-1 fraction to be consistent with the molecular mass profile of GM-CSF. Furthermore the stimulatory effect of the P-1 fraction on DNA synthesis in human melanocytes was neutralized by antibodies to GM-CSF, but not to basic fibroblast growth factor or stem cell factor. Binding and proliferation assays with recombinant GM-CSF demonstrated that human melanocytes possess specific binding sites for GM-CSF(Kd 2.11 nM; binding sites, 2.5-3.5 x 10(4) per cell), and recombinant GM-CSF at concentrations of more than 10 nM significantly stimulated DNA synthesis and melanization. These findings suggest that GM-CSF secreted by keratinocytes plays an essential role in the maintenance of melanocyte proliferation and UVA-induced pigmentation in the epidermis.

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Selected References

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  • Hearing VJ, Ekel TM. Mammalian tyrosinase. A comparison of tyrosine hydroxylation and melanin formation. Biochem J. 1976 Sep 1;157(3):549–557. [PMC free article] [PubMed]
  • Imokawa G, Mishima Y. Isolation and biochemical characterization of tyrosinase-rich GERL and coated vesicle in melanin synthesizing cells. Br J Dermatol. 1981 Feb;104(2):169–178. [PubMed]
  • Pawelek JM, Körner AM. The biosynthesis of mammalian melanin. Am Sci. 1982 Mar-Apr;70(2):136–145. [PubMed]
  • LERNER AB, FITZPATRICK TB. Biochemistry of melanin formation. Physiol Rev. 1950 Jan;30(1):91–126. [PubMed]
  • Imokawa G, Mishima Y. Loss of melanogenic properties in tyrosinases induced by glucosylation inhibitors within malignant melanoma cells. Cancer Res. 1982 May;42(5):1994–2002. [PubMed]
  • Erickson KL, Montagna W. The induction of melanogenesis by ultraviolet light in the pigmentary system of Rhesus monkeys. J Invest Dermatol. 1975 Sep;65(3):279–284. [PubMed]
  • Blog FB, Szabo G. The effects of psoralen and UVA (PUVA) on epidermal melanocytes of the tail in C57BL mice. J Invest Dermatol. 1979 Dec;73(6):533–537. [PubMed]
  • Jimbow K, Uesugi T. New melanogenesis and photobiological processes in activation and proliferation of precursor melanocytes after UV-exposure: ultrastructural differentiation of precursor melanocytes from Langerhans cells. J Invest Dermatol. 1982 Feb;78(2):108–115. [PubMed]
  • Imokawa G, Kawai M, Mishima Y, Motegi I. Differential analysis of experimental hypermelanosis induced by UVB, PUVA, and allergic contact dermatitis using a brownish guinea pig model. Arch Dermatol Res. 1986;278(5):352–362. [PubMed]
  • Imokawa G, Kawai M. Differential hypermelanosis induced by allergic contact dermatitis. J Invest Dermatol. 1987 Dec;89(6):540–546. [PubMed]
  • Halaban R, Langdon R, Birchall N, Cuono C, Baird A, Scott G, Moellmann G, McGuire J. Basic fibroblast growth factor from human keratinocytes is a natural mitogen for melanocytes. J Cell Biol. 1988 Oct;107(4):1611–1619. [PMC free article] [PubMed]
  • Gordon PR, Mansur CP, Gilchrest BA. Regulation of human melanocyte growth, dendricity, and melanization by keratinocyte derived factors. J Invest Dermatol. 1989 Apr;92(4):565–572. [PubMed]
  • Yaar M, Gilchrest BA. Human melanocyte growth and differentiation: a decade of new data. J Invest Dermatol. 1991 Oct;97(4):611–617. [PubMed]
  • Imokawa G, Yada Y, Miyagishi M. Endothelins secreted from human keratinocytes are intrinsic mitogens for human melanocytes. J Biol Chem. 1992 Dec 5;267(34):24675–24680. [PubMed]
  • Yada Y, Higuchi K, Imokawa G. Effects of endothelins on signal transduction and proliferation in human melanocytes. J Biol Chem. 1991 Sep 25;266(27):18352–18357. [PubMed]
  • Imokawa G, Yada Y, Okuda M. Allergic contact dermatitis releases soluble factors that stimulate melanogenesis through activation of protein kinase C-related signal-transduction pathway. J Invest Dermatol. 1992 Oct;99(4):482–488. [PubMed]
  • Isseroff RR, Fusenig NE, Rifkin DB. Plasminogen activator in differentiating mouse keratinocytes. J Invest Dermatol. 1983 Apr;80(4):217–222. [PubMed]
  • Whittaker JR. Biosynthesis of a thouracil pheomelanin in embryonic pigment cells exposed to thouracil. J Biol Chem. 1971 Oct 25;246(20):6217–6226. [PubMed]
  • Farishian RA, Whittaker JR. Phenylalanine lowers melanin synthesis in mammalian melanocytes by reducing tyrosine uptake: implications for pigment reduction in phenylketonuria. J Invest Dermatol. 1980 Feb;74(2):85–89. [PubMed]
  • Oikawa A, Nakayasu M, Nohara M, Tchen TT. Fate of L-(3,5- 3H) tyrosine in cell-free extracts and tissue cultures of melanoma cells: a new assay method for tyrosinase in living cells. Arch Biochem Biophys. 1972 Feb;148(2):548–557. [PubMed]
  • Imokawa G, Motegi I. Skin organ culture model for examining epidermal melanization. J Invest Dermatol. 1993 Jan;100(1):47–54. [PubMed]
  • Eisinger M, Marko O. Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2018–2022. [PMC free article] [PubMed]
  • Morelli JG, Yohn JJ, Lyons MB, Murphy RC, Norris DA. Leukotrienes C4 and D4 as potent mitogens for cultured human neonatal melanocytes. J Invest Dermatol. 1989 Dec;93(6):719–722. [PubMed]
  • Imokawa G, Tejima T. A possible role of prostaglandins in PUVA-induced inflammation: implication by organ cultured skin. J Invest Dermatol. 1989 Feb;92(2):296–300. [PubMed]
  • Imokawa G, Miyagishi M, Yada Y. Endothelin-1 as a new melanogen: coordinated expression of its gene and the tyrosinase gene in UVB-exposed human epidermis. J Invest Dermatol. 1995 Jul;105(1):32–37. [PubMed]
  • Gahring L, Baltz M, Pepys MB, Daynes R. Effect of ultraviolet radiation on production of epidermal cell thymocyte-activating factor/interleukin 1 in vivo and in vitro. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1198–1202. [PMC free article] [PubMed]
  • Metcalf D. The granulocyte-macrophage colony-stimulating factors. Science. 1985 Jul 5;229(4708):16–22. [PubMed]
  • Clark SC, Kamen R. The human hematopoietic colony-stimulating factors. Science. 1987 Jun 5;236(4806):1229–1237. [PubMed]
  • Bagby GC, Jr, McCall E, Bergstrom KA, Burger D. A monokine regulates colony-stimulating activity production by vascular endothelial cells. Blood. 1983 Sep;62(3):663–668. [PubMed]
  • Lee F, Yokota T, Otsuka T, Gemmell L, Larson N, Luh J, Arai K, Rennick D. Isolation of cDNA for a human granulocyte-macrophage colony-stimulating factor by functional expression in mammalian cells. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4360–4364. [PMC free article] [PubMed]
  • Gallo RL, Grabbe S, Choi SS, Bleicher P, Granstein RD. Cyclosporin increases granulocyte/macrophage colony-stimulating factor (GM-CSF) activity and gene expression in murine keratinocytes. J Invest Dermatol. 1992 Mar;98(3):274–278. [PubMed]
  • Kupper TS, Lee F, Birchall N, Clark S, Dower S. Interleukin 1 binds to specific receptors on human keratinocytes and induces granulocyte macrophage colony-stimulating factor mRNA and protein. A potential autocrine role for interleukin 1 in epidermis. J Clin Invest. 1988 Nov;82(5):1787–1792. [PMC free article] [PubMed]
  • Gallo RL, Staszewski R, Sauder DN, Knisely TL, Granstein RD. Regulation of GM-CSF and IL-3 production from the murine keratinocyte cell line PAM 212 following exposure to ultraviolet radiation. J Invest Dermatol. 1991 Aug;97(2):203–209. [PubMed]
  • Nozaki S, Abrams JS, Pearce MK, Sauder DN. Augmentation of granulocyte/macrophage colony-stimulating factor expression by ultraviolet irradiation is mediated by interleukin 1 in Pam 212 keratinocytes. J Invest Dermatol. 1991 Jul;97(1):10–14. [PubMed]

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