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  • Title: Phospholipid composition of human monocytes and alterations occurring due to culture and stimulation by C3b.
    Author: Kennett FF, Schenkein HA, Ellis TM, Rutherford RB.
    Journal: Biochim Biophys Acta; 1984 Jul 20; 804(3):301-7. PubMed ID: 6743692.
    Abstract:
    The phospholipid composition of human peripheral blood monocytes has not been previously reported, due to difficulty in isolating these cells in a purified state. In this study, monocytes were purified by counterflow centrifugation without selective adherence, and were characterized with the use of fluorescent monoclonal antibodies to T and B lymphocytes and monocytes by flow cytometry. These platelet-free cell preparations contained less than 5% T cells and less than 3% B cells. Isolated monocytes, which were rapidly frozen after isolation, contained phospholipids (in order of decreasing concentrations) as follows: phosphatidylcholine greater than phosphatidylethanolamine greater than sphingomyelin greater than phosphatidylserine greater than phosphatidylinositol greater than cardiolipin. A small amount of lyso-PC, but no lyso-PE, phosphatidic acid or lyso-PI, was found. The effect of culturing these cells in the presence or absence of a known stimulant of monocyte prostaglandin E and thromboxane release, the C3b fragment of the third component of human complement (C3), was studied with regard to phospholipid composition. Monocytes cultured without stimulant for 24 h contained 3-4% more sphingomyelin than did uncultured cells, and lyso-PC concentrations were consistently elevated. The addition of the stimulant C3b to cultured cells resulted in enhancement of release of immunoreactive prostaglandin E into culture supernatants, without affecting the release of lysosomal enzymes. Analysis of the phospholipid content of cells cultured in the presence of C3b revealed that there was a significant decrease in total PI compared to cells cultured in the absence of C3b, in addition to an increased concentration of sphingomyelin and lyso-PC when compared to freshly isolated cells. These changes occurred in the absence of elevated concentrations of phosphatidic acid.
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