비접촉 조건에서의 Naegleria fowleri에 의한 표적세포의 세포독성
Cytotoxicity of target cell against Naegleria fowleri under non-contact condition
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초록
Naegleria fowleri, a pathogenic free-living amoeba, leads to a fatal infection known as primary amebic meningoencephalitis (PAM) in human and animals. PAM is an acute, fulminant, necrotizing, and hemor-rhagic disease that leads to death in approximately seven days. In this study, we investigate the cytotox-icity of target cells and the secreted molecules of N. fowleri under the non-contact condition. The target cell (U87MG cell) treated with N. fowleri lysates showed no morphological changes and no cytotoxicity. By contrast, the U87MG cells co-cultured with N. fowleri trophozoites under the non-contact condition induced morphological changes and reduction in number. When U87MG cells were co-cultured with N. fowleri trophozoites under the non-contact condition for 30 min, 2 hr, and 4 hr, the levels of cytotox-icity of target cells were 32.3, 35.5, and 37.8%, respectively. Particularly, when the ratio of amoeba to target cells is 10 to 1, the level of cytotoxicity of target cells was 49.7% at 30 min. To show the proteins secreted from N. fowleri under the non-contact condition, we carried out 2D electrophoresis and observed 6 major proteins. Finally, these results suggest that the molecules released from N. fowleri un-der the non-contact condition induce the cell death and this process is an important step in pathogenesis of N. fowleri.
Abstract
Naegleria fowleri, a pathogenic free-living amoeba, leads to a fatal infection known as primary amebic meningoencephalitis (PAM) in human and animals. PAM is an acute, fulminant, necrotizing, and hemor-rhagic disease that leads to death in approximately seven days. In this study, we investigate the cytotox-icity of target cells and the secreted molecules of N. fowleri under the non-contact condition. The target cell (U87MG cell) treated with N. fowleri lysates showed no morphological changes and no cytotoxicity. By contrast, the U87MG cells co-cultured with N. fowleri trophozoites under the non-contact condition induced morphological changes and reduction in number. When U87MG cells were co-cultured with N. fowleri trophozoites under the non-contact condition for 30 min, 2 hr, and 4 hr, the levels of cytotox-icity of target cells were 32.3, 35.5, and 37.8%, respectively. Particularly, when the ratio of amoeba to target cells is 10 to 1, the level of cytotoxicity of target cells was 49.7% at 30 min. To show the proteins secreted from N. fowleri under the non-contact condition, we carried out 2D electrophoresis and observed 6 major proteins. Finally, these results suggest that the molecules released from N. fowleri un-der the non-contact condition induce the cell death and this process is an important step in pathogenesis of N. fowleri.
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