Identification and pharmacological characterization of platelet-activating factor and related 1-palmitoyl species in human inflammatory blistering diseases.
Through its pro-inflammatory effects on leukocytes, endothelial cells, and keratinocytes, the lipid mediator platelet-activating factor (PAF) has been implicated in cutaneous inflammation. Although the 1-alkyl PAF species has been considered historically the most abundant and important ligand for the PAF receptor (PAF-R), other putative ligands for this receptor have been described including 1-acyl analogs of sn-2 acetyl glycerophosphocholines. Previous bioassays have demonstrated a PAF-like activity in lesions of the autoimmune blistering disease bullous pemphigoid. To assess the actual sn-2 acetyl glycerophosphocholine species that result in this PAF agonistic activity, we measured PAF and related sn-2 acetyl GPCs in fresh blister fluid samples from bullous pemphigoid and noninflammatory (suction-induced) bullae by mass spectrometry. We report the presence of 1-hexadecyl as well as the 1-acyl PAF analog 1-palmitoyl-2-acetyl glycerophosphocholine (PAPC) in inflammatory blister fluid samples. Because PAPC is the most abundant sn-2 acetyl glycerophosphocholine species found in all samples examined, the pharmacological effects of this species with respect to the PAF-R were determined using a model system created by transduction of a PAF-R-negative epidermoid cell line with the PAF-R. Radioligand binding and intracellular calcium mobilization studies indicated that PAPC is approximately 100x less potent than PAF. Though a weak agonist, PAPC could induce PAF biosynthesis and PAF-R desensitization. Finally, intradermal injections of PAF and PAPC into the ventral ears of rats demonstrated that PAPC was 100x less potent in vivo. These studies suggest possible involvement of PAF and related species in inflammatory bullous diseases.
Travers
J B
JB
Department of Dermatology, Indiana University School of Medicine, Indianapolis 46202, USA. jtravers@wpo.iupui.edu
Murphy
R C
RC
Johnson
C A
CA
Pei
Y
Y
Morin
S M
SM
Clay
K L
KL
Barber
L A
LA
Hood
A F
AF
Morelli
J G
JG
Williams
D A
DA
D000818:Animals; D001667:Binding, Competitive; D001768:Blister / Q000378:metabolism; D015152:Blotting, Northern; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis; D005997:Glycerylphosphorylcholine / Q000031:analogs & derivatives* / Q000032:analysis; D006801:Humans; D007249:Inflammation / Q000139:chemically induced; D007624:KB Cells; D010391:Pemphigoid, Bullous / Q000378:metabolism*; D010713:Phosphatidylcholines; D010742:Phospholipid Ethers / Q000032:analysis; D010972:Platelet Activating Factor / Q000819:agonists* / Q000031:analogs & derivatives / Q000235:genetics / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae; D013997:Time Factors; D014161:Transduction, Genetic