Identification of functional platelet-activating factor receptors on human keratinocytes

Jeffrey Travers, J. C. Huff, M. Rola-Pleszczynski, E. W. Gelfand, J. G. Morelli, R. C. Murphy

Research output: Contribution to journalArticle

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Abstract

Platelet-activating factor (PAF) is a potent inflammatory mediator that has been shown to be produced by human keratinocytes and is thought to play a role in cutaneous inflammation. Immunofluorescence and radioligand binding studies were used to characterize PAF receptors (PAF-R) on human keratinocytes and the human epidermoid cell lines A-431 and HaCaT. Indirect immunofluorescence studies demonstrated anti-PAF-R staining of primary cultures of human keratinocytes, A-431 cells, and HaCaT cells. Primary cultures of human fibroblasts and the melanoma cell line SK-30 failed to show immunostaining above that seen with control antiserum. With indirect immunofluorescence studies of sections of normal human skin, a granular anti-PAF-R staining pattern was noted on the keratinocyte cell membranes. A-431 cells readily metabolized PAF by deacetylation-reacylation at 37°C, but not at 4°C. Binding studies on crude membrane preparations of A-431 cells conducted at 4°C demonstrated specific binding that reached saturation by 120 min. Scatchard analysis of PAF binding data revealed a single class of high-affinity (K(D) = 6.3 ± 0.3 nM) PAF binding sites. The immunofluorescence and radioligand binding sites were shown to be functional PAF-Rs, as 10 pM to 1 μM PAF increased intracellular calcium in primary cultures of human keratinocytes, A-431 cells, and HaCaT cells, whereas PAF treatment of primary cultures of human fibroblasts or the melanoma cell line SK-30 did not result in changes in the intracellular calcium concentration. The structurally dissimilar PAF-R antagonists CV-6209, Ro19-3704, and alprazolam all inhibited the PAF-induced calcium changes in A-431 cells. The CV-6209 inhibition was seen at doses that competed with the PAF binding to these cells. These studies provide the first evidence for the presence of a functional PAF-R expressed on human keratinocytes, suggesting that this lipid mediator may play an important role in normal keratinocytes or in inflammatory dermatology.

Original languageEnglish
Pages (from-to)816-823
Number of pages8
JournalJournal of Investigative Dermatology
Volume105
Issue number6
StatePublished - 1995

Fingerprint

Platelet Activating Factor
Keratinocytes
Cells
Fibroblasts
Indirect Fluorescent Antibody Technique
Calcium
Cell culture
Cell Line
platelet activating factor receptor
Fluorescent Antibody Technique
Melanoma
Binding Sites
Dermatology
Staining and Labeling
Alprazolam
Skin
Cell membranes
Immune Sera

ASJC Scopus subject areas

  • Dermatology

Cite this

Travers, J., Huff, J. C., Rola-Pleszczynski, M., Gelfand, E. W., Morelli, J. G., & Murphy, R. C. (1995). Identification of functional platelet-activating factor receptors on human keratinocytes. Journal of Investigative Dermatology, 105(6), 816-823.

Identification of functional platelet-activating factor receptors on human keratinocytes. / Travers, Jeffrey; Huff, J. C.; Rola-Pleszczynski, M.; Gelfand, E. W.; Morelli, J. G.; Murphy, R. C.

In: Journal of Investigative Dermatology, Vol. 105, No. 6, 1995, p. 816-823.

Research output: Contribution to journalArticle

Travers, J, Huff, JC, Rola-Pleszczynski, M, Gelfand, EW, Morelli, JG & Murphy, RC 1995, 'Identification of functional platelet-activating factor receptors on human keratinocytes', Journal of Investigative Dermatology, vol. 105, no. 6, pp. 816-823.
Travers J, Huff JC, Rola-Pleszczynski M, Gelfand EW, Morelli JG, Murphy RC. Identification of functional platelet-activating factor receptors on human keratinocytes. Journal of Investigative Dermatology. 1995;105(6):816-823.
Travers, Jeffrey ; Huff, J. C. ; Rola-Pleszczynski, M. ; Gelfand, E. W. ; Morelli, J. G. ; Murphy, R. C. / Identification of functional platelet-activating factor receptors on human keratinocytes. In: Journal of Investigative Dermatology. 1995 ; Vol. 105, No. 6. pp. 816-823.
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