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Quantitation by radioimmunoassay of PAF in human saliva

  • PAF Analysis by Spectrometric and Immunological Assay
  • Published:
Lipids

Abstract

Approximately 75% of the PAF present in saliva is recovered on extraction of whole saliva (0.8 vol) with chloroform/methanol/water (2∶2∶1 v/v/v). PAF levels, determined by our recently developed radioimmunoassay, in saliva extracts ranged from 0.5–21 ng/mL with 59% between 2–6 ng/mL. These figures, for apparently healthy subjects, are higher than previously reported levels obtained by platelet assays. The validity of our radioimmunoassay results was checked by isolating and quantitating the PAF fraction from whole saliva. In addition, when we examined our saliva samples by platelet aggregation, low levels of PAF, comparable with the values found in the literature, were detected. Investigations revealed the presence of a substance(s) which inhibited PAF-induced platelet aggregation but which did not affect the radioimmunoassay.

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Abbreviations

AcT:

0.05M acetate buffer pH 6.0 with 0.05% Tween-20

B0 :

binding to antibody in absence of inhibitor

BSA:

bovine serum albumin

CnPAF:

PAF where the 1-O-alkyl chain is n carbons long

[3H-acetyl]PAF:

1-O-alkyl-2-3H]acetyl-sn-glycero-3-phosphocholine

[3H-alkyl]PAF:

1-O-[3H]alkyl-2-acetyl-sn-glycero-3-phosphocholine

Ig:

immunoglobulin

[125I]PAF:

1-O-[125I]alkyl-2-acetyl-sn-glycero-3-phosphocholine

lyso-PAF:

1-O-alkyl-sn-glycero-3-phosphocholine

NSB:

nonspecific binding

PAF:

1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine

PBS:

phosphate (50 mM) buffered saline pH 7.2

PLA2 :

phospholipase A2

RIA:

radioimmunoassay

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Authors and Affiliations

  1. Kolling Institute of Medical Research, Royal North Shore Hospital, St., 2065, Leonards, N.S.W.

    Sue J. Cooney, Mary A. Smal & Brian A. Baldo

  2. Department of Medicine, University of Sydney, 2006, N.S.W., Australia

    Brian A. Baldo

Authors
  1. Sue J. Cooney
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  2. Mary A. Smal
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  3. Brian A. Baldo
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Cooney, S.J., Smal, M.A. & Baldo, B.A. Quantitation by radioimmunoassay of PAF in human saliva. Lipids 26, 1140–1143 (1991). https://doi.org/10.1007/BF02536518

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  • DOI: https://doi.org/10.1007/BF02536518

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