Mirzaei Farzaneh, Khanahmad Hossein, Namdar Fatemeh, Izadi Shahrokh, Hejazi Seyed Hossein
Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Iran J Parasitol. 2019 Oct-Dec;14(4):542-551.
We aimed to design a different method of drug delivery for increased transfer of the choice drug (meglumine antimoniate) within the host cells. Therefore, listeriolysin O (LLO), a bacterial product which is a member of pore-forming peptides was used as an enhancer factor with meglumine antimoniate in order to facilitate the transition of the drug across macrophage membrane.
LLO was produced in Isfahan University of Medical Sciences in 2016, by expressing the gene in and purified using affinity chromatography. Cytotoxicity of the purified protein was investigated in an in vitro model of macrophage infection.
LLO was cytotoxic against murine macrophage cells (J774-A1) and amastigote forms of (MRHO/IR/75/ER). It was less toxic to macrophages (CC50=2.56 μg ml ±0.09) than to the parasites (IC50=1.72 μg ml ±0.07). Moreover, noncytotoxic concentration of LLO (0.006 ug ml) potentiated the cytotoxicity induced by low dose concentration of meglumine antimoniate. Very little dose of meglumine antimoniate was needed when combined with the LLO (IC50=12.63 μg ml ±0.13) in comparison with the cytotoxicity induced when the drug is used alone (IC50=46.17 μg ml ±0.28).
The combination of pore-forming proteins with anti-leishmanial agents could increase the advantage of anti-leishmanial drugs. Since lower concentrations of anti-leishmanial drugs can reduce undesirable side effects of chemotherapy trials carried out in animal models and then in humans with this system.
我们旨在设计一种不同的药物递送方法，以增加宿主细胞内所选药物（葡甲胺锑酸盐）的转运。因此，将成孔肽家族成员之一的细菌产物李斯特菌溶素O（LLO）与葡甲胺锑酸盐一起用作增强因子，以促进药物穿过巨噬细胞膜的转运。
2016年在伊斯法罕医科大学通过在大肠杆菌中表达该基因来生产LLO，并使用亲和色谱法进行纯化。在巨噬细胞感染的体外模型中研究纯化蛋白的细胞毒性。
LLO对小鼠巨噬细胞（J774-A1）和杜氏利什曼原虫的无鞭毛体形式（MRHO/IR/75/ER）具有细胞毒性。它对巨噬细胞的毒性（CC50=2.56μg/ml±0.09）低于对寄生虫的毒性（IC50=1.72μg/ml±0.07）。此外，LLO的无细胞毒性浓度（0.006μg/ml）增强了低剂量葡甲胺锑酸盐诱导的细胞毒性。与单独使用该药物时诱导的细胞毒性（IC50=46.17μg/ml±0.28）相比，与LLO联合使用时所需的葡甲胺锑酸盐剂量非常小（IC50=12.63μg/ml±0.13）。
成孔蛋白与抗利什曼原虫药物的组合可以增加抗利什曼原虫药物的优势。因为较低浓度的抗利什曼原虫药物可以减少在动物模型以及随后在人体中进行化疗试验时产生的不良副作用。
