Fas and Ley antigen expression in polymorphonuclear leukocytes. Flow cytometric analysis.:flow cytometric analysis
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Recently, it has been suggested that apoptosis of polymorphonuclear leukocytes (PMNs) plays a role in the regulation of inflammation (e. g., periodontitis). The present study investigated the relationship between cytokine produced during the course of inflammation and the apoptosis induction of PMNs. Peripheral blood PMNs (BPMNs) and salivary PMNs (SPMNs) were isolated from eight volunteers and treated with PBS, non heat-treated saliva, heat-treated saliva, TNF-α or PMA for 0, 3 hours at 37°C. The sample was examined for apoptosis receptor expression and apoptosis expression on cell surfaces by indirect immunofluorescence using anti-human Fas monoclonal antibody and anti-human Le<SUP>y</SUP> monoclonal antibody. The fluorescence intensity of 5, 000 cells in each sample was measured using a flow cytometer and the percentage of FITC positive cells was calculated. The % of Fas-positive cells were almost the same in BPMNs and SPMNs at 0h. In BPMNs, saliva, TNF-α and Fas extremely increased Fas receptor expression on cell surfaces at 3h. The % of Le<SUP>y</SUP>-positive cells was higher in SPMNs than in BPMNs at 0h. In BPMNs, saliva, TNF-α and Fas increased Le<SUP>y</SUP> antigen expression at 3h, but heated saliva did not it. In SPMNs, Le<SUP>y</SUP> antigen expression was not increased in PBS and the serum group at 3h compared with that at 0h. These findings suggest that apoptosis of PMN may be induced by cytokines such as TNF-α in periodental tissue and induced by stimulation of saliva in the oral cavity.
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