An optical and chemiluminescence assay for assessing the cy-totoxicity of Balamuthia mandrillaris against human neuro-spheroids

<p>A spheroid is a cell aggregate in a three-dimensional context; thereby, it recapitulates the cellular architecture in human tissue. However, the utility of spheroids as an assay for host-parasite interactions remains unexplored. This study demonstrates the potential use of neurospheroids for assessing the cytotoxicity of the life-threatening pathogenic amoeba <em>Balamuthia mandrillaris</em>. The neuroblastoma SH-SY5Y cells formed a spheroid in a hanging drop of culture medium. Cellular damage of <em>B. mandrillaris</em> trophozoites on human neuronal spheroids was observed using microscopic imaging and ATP detection. <em>B. mandrillaris</em> trophozoites rapidly caused a decrease in ATP production in the spheroid, leading to loss of neurospheroid integrity. Moreover, 3D confocal microscopy imaging revealed interactions between the trophozoites and SH-SY5Y neuronal cells in the outer layer of the neurospheroid. In conclusion, the neurospheroid allows the assessment of host cell damage in a simple and quantitative manner.</p>