In the present study, to determine whether a pathogenic N. fowleri lysate shows the cytopathic effects against primary culture rat microglial cells, the morphological changes of microglial cells was observed by a light, scanning and transmission electron microscope.
And then, the cytotoxicity of N. fowleri lysate against microglial cells was also observed by
51Cr release assay.
In addition, the pro-inflammatory cytokine release from microglial cells in the co-culture system was estimated. As results with a light and electron microscopes, most of microglial cells were severely destroyed by N. fowleri lysate, showing the necrotic (above 85%) and apoptotic cell death (below 15%) in a time- and dose dependent manner.
As the results of 51Cr release assay, the cytotoxicity of N. fowleri lysate against microglial cells were 14.6, 21.9, 38.5 and 71.5% at 3, 6, 12 and 24 hr post incubation, respectively. And then, the amount of cytokines released from microglial cells in co-culture systems at 3, 6 and 12 hr were 121.6, 90.4 and 81.0 pg/ml of TNF-α, 88.5, 92.7 and 190.1 pg/ml of IL-1β, and 298.8, 414.9 and 251.1 pg/ml of IL-6, respectively.
Thus, this present study shows that the microglial cells may be involved in the inflammation stage of N. fowleri infection as secreting various inflammatory cytokines.
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- 국문요약 -
cell sorting(FACS)을 시행한 결과, 6시간 후 약 16% 정도가 세포사멸 (apoptosis) 과정을 통해 사멸하였다. 또한, 표적세포인 신경소교세포에 대한 아 메바 세포추출액의 세포독성을 생화학적 방법인 51Cr 분비 실험으로 관찰한 결 과, 신경소교세포들에 대한 자유아메바의 세포독성은 배양 후 3, 6, 12 그리고 24 시간에 각각 14.6, 21.9, 38.5 그리고 71.5% 이었다.
한편, 파울러자유아메바의 세포 추출액과 혼합 배양한 신경소교세포들은 방어 적인 면역반응으로 다양한 싸이토카인들을 분비하였는데, TNF-α 및 IL-6등의 싸이토카인(cytokine) 분비량은 배양 초기부터 증가하였으며, IL-1β의 분비는 12시간 후부터 증가하였다.
핵심어: 파울러자유아메바, 원발성 아메바성 수막뇌염, 신경소교세포, 세포독성, 싸이토카인