The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, Israel.
Toxins (Basel). 2019 Mar 22;11(3):174. doi: 10.3390/toxins11030174.
Ricin and abrin are ribosome-inactivating proteins leading to inhibition of protein synthesis and cell death. These toxins are considered some of the most potent and lethal toxins against which there is no available antidote. Digital holographic microscopy (DHM) is a time-lapse, label-free, and noninvasive imaging technique that can provide phase information on morphological features of cells. In this study, we employed DHM to evaluate the morphological changes of cell lines during ricin and abrin intoxication. We showed that the effect of these toxins is characterized by a decrease in cell confluence and changes in morphological parameters such as cell area, perimeter, irregularity, and roughness. In addition, changes in optical parameters such as phase-shift, optical thickness, and effective-calculated volume were observed. These effects were completely inhibited by specific neutralizing antibodies. An enhanced intoxication effect was observed for preadherent compared to adherent cells, as was detected in early morphology changes and confirmed by annexin V/propidium iodide (PI) apoptosis assay. Detection of the dynamic changes in cell morphology at initial stages of cell intoxication by DHM emphasizes the highly sensitive and rapid nature of this method, allowing the early detection of active toxins.
蓖麻毒素和相思豆毒素是核糖体失活蛋白，可导致蛋白质合成抑制和细胞死亡。这些毒素被认为是针对其尚无有效解毒剂的最有效和最致命的毒素之一。数字全息显微镜（DHM）是一种无标记、非侵入性的实时成像技术，可提供细胞形态特征的相位信息。在这项研究中，我们采用 DHM 来评估细胞系在蓖麻毒素和相思豆毒素中毒时的形态变化。结果表明，这些毒素的作用特征是细胞融合减少以及细胞面积、周长、不规则性和粗糙度等形态参数发生变化。此外，还观察到光学参数如相移、光学厚度和有效计算体积的变化。这些效应均被特异性中和抗体完全抑制。与贴壁细胞相比，预贴壁细胞的中毒效应增强，这在早期形态变化中被检测到，并通过 Annexin V/碘化丙啶（PI）凋亡检测得到证实。DHM 对细胞中毒初始阶段细胞形态动态变化的检测强调了该方法的高灵敏度和快速性，可早期检测到活性毒素。
