Laboratório de Bacteriologia, Instituto Butantan, São Paulo 05503-900, SP, Brazil.
Laboratório de Toxinologia Aplicada, Instituto Butantan, São Paulo 05503-900, SP, Brazil.
Toxins (Basel). 2021 Jun 12;13(6):419. doi: 10.3390/toxins13060419.
Heat-labile toxin I (LT-I), produced by strains of enterotoxigenic (ETEC), causes profuse watery diarrhea in humans. Different in vitro and in vivo models have already elucidated the mechanism of action of this toxin; however, their use does not always allow for more specific studies on how the LT-I toxin acts in systemic tracts and intestinal cell lines. In the present work, zebrafish () and human intestinal cells (Caco-2) were used as models to study the toxin LT-I. Caco-2 cells were used, in the 62nd passage, at different cell concentrations. LT-I was conjugated to FITC to visualize its transport in cells, as well as microinjected into the caudal vein of zebrafish larvae, in order to investigate its effects on survival, systemic traffic, and morphological formation. The internalization of LT-I was visualized in 3 × 10 Caco-2 cells, being associated with the cell membrane and nucleus. The systemic traffic of LT-I in zebrafish larvae showed its presence in the cardiac cavity, yolk, and regions of the intestine, as demonstrated by cardiac edema (100%), the absence of a swimming bladder (100%), and yolk edema (80%), in addition to growth limitation in the larvae, compared to the control group. There was a reduction in heart rate during the assessment of larval survival kinetics, demonstrating the cardiotoxic effect of LT-I. Thus, in this study, we provide essential new depictions of the features of LT-I.
不耐热肠毒素 I(LT-I)由肠致病性(ETEC)菌株产生,可导致人类大量水样腹泻。不同的体外和体内模型已经阐明了这种毒素的作用机制;然而,它们的使用并不总是允许对 LT-I 毒素在全身和肠道细胞系中的作用进行更具体的研究。在本工作中,斑马鱼()和人肠细胞(Caco-2)被用作研究 LT-I 毒素的模型。使用处于第 62 代的不同细胞浓度的 Caco-2 细胞。将 LT-I 缀合到 FITC 上,以可视化其在细胞中的转运,并用微注射到斑马鱼幼虫的尾静脉中,以研究其对存活率、全身运输和形态形成的影响。在 3×10 个 Caco-2 细胞中观察到 LT-I 的内化,与细胞膜和细胞核相关。LT-I 在斑马鱼幼虫中的全身运输显示其存在于心房、卵黄和肠道区域,如心脏水肿(100%)、无鳔(100%)和卵黄水肿(80%),以及与对照组相比,幼虫生长受限。在评估幼虫存活动力学时,心率下降,证明了 LT-I 的心脏毒性作用。因此,在这项研究中,我们提供了 LT-I 特征的重要新描述。