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KMID : 0357119930150020191
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Korean Journal of Immunology
1993 Volume.15 No. 2 p.191 ~ p.199
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The Effect of a Factor Produced from Phytohaemagglutinin-stimulated T Cells on the Neutrophil Viability, Superoxide Production and Leukotriene C₄Release
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Her Erk
Yang Young-Mok
Yang Young-Mok
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Abstract
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here are several direct and indirect ways in which T cells could enhance the anti-bacterial capabilities of neutrophils. However it is not yet clear out which molecule or cytokine produced by T cells is involved in the phagocytic action and viability sustaining activity of neutrophils. The aim of this study was about a factor, which produced by the phytohaemagglutinin(PHA)-stimulated T cells, may control those neutrophil actions. Human peripheral blood T cells and neutrophils were isolated by Ficoll-paque density sedimentation from heparinized blood of healthy adult donors. The purity of these cells were more than 90%. T cells were stimulated in various dose(0.1-10gg/ml) of PHA for various times of incubation(0-3 days), and then PHA-stimulated T cell conditioned medium was collected in order to find an optimal dose and incubation time for the neutrophil viability. It was found out that 1,ug/ml of PHA in 12 hours incubation was maximal effective condition for the neutrophil sustaining viability. The effects of PHA-stimulated T cell conditioned medium(TCM) on the neutrophils were used for the comparison with PHA-nonstimulated TCM or enriched medium alone. Neutrophil sustaining viability with PHA-stimulated TCM for 24 hours incubation was significantly higher than other groups (80 ± 10 vs 25 ±15 vs 13 ± 9; p <0.01). The superoxide prodution from neutrophils with PHA-stimulated TCM for 24 hours incubation were also significantly higher than other groups(25±4 vs 11±4 vs 7±5; p <0.01). In the leukotriene C4 (LTC4) release, neutrophils with PHA-stimulated TCM for 24 hours incubation were different from other group (105 ± 20 vs 65 ±1O vs 25 ± 32 ; p <0.01), and unlikely other parameters the cells with PHA-nonstimulated TCM was different from the cells with enriched medium alone(65 -1:10 vs 25-± 32; p<0.05). In two dimension electrophoresis it was shown that PHA-stimulated T cells enhanced three proteins(66kD, 60kD, 45kD) and diminished one(40kD) in the production and/or release of proteins in comparison with PHA-nonstimulated T cells. These data suggest that these proteins from the PHA-stimulated T cells might be involved in the phagocytic actions of neutrophils.
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KEYWORD
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Neutrophil, Phytohaemagglutinin(PHA), T cell-conditioned medium
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