Primary cultivated amnion epitherial cells prepared from amnion tissue of human fetal membrane (Amnion-cells) were stimulated with influenza virus-hemagglutinin (IV-HA), fractionated from a commercialized IV-HA vaccine by DEAE Sephacel column chromatography. From 72—96 h after stimulation, chromosomal DNA fragmentation and the appearance of in situ TUNEL stained-positive cells were revealed. Amnion-cell DNA fragmentation was inhibited in the presence of glycophorin A or C purified from the human erythrocyte membrane fraction, but not inhibited with free N-acetyl-neuraminic acid. RT-PCR and Western blotting analysis showed that anti-oxidative enzymes were altered with incubation period, accompanied by the expression of the cellular oxidative stress-related caspase cascade. Pre-stimulation of Amnion-cells with hemin, a heme oxygenase-1 inducer, significantly attenuated IV-HA induced DNA fragmentation. It is concluded that IV-HA induces apoptosis in Amnion-cells, and that this apoptotic induction may be facilitated by certain sialoglycoproteins on the cell surface, and is related to changes in the intracellular redox condition.
2003 The Pharmaceutical Society of Japan