Platelet-activating factor biosynthesis induced by various stimuli in human HaCaT keratinocytes

Jeffrey Travers, Kathleen A. Harrison, Christopher A. Johnson, Keith L. Clay, Joseph G. Morelli, Robert C. Murphy

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Platelet-activating factor (PAF) is a potent inflammatory mediator that is thought to play a role in cutaneous inflammation. These studies used mass spectrometry to examine the molecular species of PAF precursor glycerophosphocholine lipids (GPC) as well as the biosynthesis of PAF and other sn-2 acetyl-GPC in a human keratinocyte-derived cell line (HaCaT keratinocytes). Approximately 28% of HaCaT keratinocyte GPC consisted of 1-alkyl species, and the relative amounts of the sn-1 alkyl constituents of the PAF precursor 1-alkyl-2-acyl-GPC were as follows: hexadecyl > octadecenyl > octadecyl. Ionophore (A23187)-stimulated HaCaT keratinocytes synthesized both PAF (1-hexadecyl, 1-octadecenyl, and 1-octadecyl species) and less potent 1-acyl analogs (1-palmitoyl, 1-oleoyl, and 1-stearoyl species). PAF production was rapid and maximal by 10 min. The major species of sn-2 acetyl-GPC at 2.5 min were 1-hexadecyl-2-acetyl-GPC (2.2 ng/106 cells) and 1-palmitoyl-2-acetyl-GPC (2.4 ng/106 cells). HaCaT keratiraocytes also synthesized PAF and 1-acyl PAF analogs when stimulated with the peptide growth factor endothelin-1 and the nonhydrolyzable PAF receptor agonist carbamyl-PAF. Both 1-hexadecyl-2-acetyl-GPC and 1-palmitoyl-2-acetyl-GPC stimulated intracellular calcium mobilization in HaCaT cells, indicating that these sn-2 acetyl-GPC act in autocrine fashion, These studies revealed that the human keratinocyte-derived cell line HaCaT can synthesize significant amounts of PAF and 1-acyl analogs in vitro from both nonspecific (A23187) and specific (endothelin-1, carbamyl-PAF) stimulation, suggesting a role for this inflammatory lipid mediator in keratinocyte pathophysiology.

Original languageEnglish
Pages (from-to)88-94
Number of pages7
JournalJournal of Investigative Dermatology
Volume107
Issue number1
StatePublished - 1996

Fingerprint

Platelet Activating Factor
Biosynthesis
Keratinocytes
Lipids
Calcimycin
Endothelin-1
Cells
Cell Line
Ionophores
Mass spectrometry
Mass Spectrometry
Intercellular Signaling Peptides and Proteins
Inflammation
Calcium

Keywords

  • Endothelin
  • Glycerophospholipids
  • Intracellular calcium
  • Mass spectrometry
  • Molecular species

ASJC Scopus subject areas

  • Dermatology

Cite this

Travers, J., Harrison, K. A., Johnson, C. A., Clay, K. L., Morelli, J. G., & Murphy, R. C. (1996). Platelet-activating factor biosynthesis induced by various stimuli in human HaCaT keratinocytes. Journal of Investigative Dermatology, 107(1), 88-94.

Platelet-activating factor biosynthesis induced by various stimuli in human HaCaT keratinocytes. / Travers, Jeffrey; Harrison, Kathleen A.; Johnson, Christopher A.; Clay, Keith L.; Morelli, Joseph G.; Murphy, Robert C.

In: Journal of Investigative Dermatology, Vol. 107, No. 1, 1996, p. 88-94.

Research output: Contribution to journalArticle

Travers, J, Harrison, KA, Johnson, CA, Clay, KL, Morelli, JG & Murphy, RC 1996, 'Platelet-activating factor biosynthesis induced by various stimuli in human HaCaT keratinocytes', Journal of Investigative Dermatology, vol. 107, no. 1, pp. 88-94.
Travers, Jeffrey ; Harrison, Kathleen A. ; Johnson, Christopher A. ; Clay, Keith L. ; Morelli, Joseph G. ; Murphy, Robert C. / Platelet-activating factor biosynthesis induced by various stimuli in human HaCaT keratinocytes. In: Journal of Investigative Dermatology. 1996 ; Vol. 107, No. 1. pp. 88-94.
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