Jeddi Fakhri, Mary Charles, Aoun Karim, Harrat Zoubir, Bouratbine Aïda, Faraut Françoise, Benikhlef Rezika, Pomares Christelle, Pratlong Francine, Marty Pierre, Piarroux Renaud
Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France
Laboratoire de Parasitologie-Mycologie, CHU Timone, UMR MD3 Aix-Marseille Université, Marseille, France.
Antimicrob Agents Chemother. 2014 Aug;58(8):4866-74. doi: 10.1128/AAC.02521-13. Epub 2014 Jun 9.
Antimonials remain the first-line treatment for the various manifestations of leishmaniasis in most areas where the disease is endemic, and increasing cases of therapeutic failure associated with parasite resistance have been reported. In this study, we assessed the molecular status of 47 clinical isolates of Leishmania causing visceral and cutaneous leishmaniasis from Algeria, Tunisia, and southern France. In total, we examined 14 genes that have been shown to exhibit significant variations in DNA amplification, mRNA levels, or protein expression with respect to resistance to antimonials. The gene status of each clinical isolate was assessed via qPCR and qRT-PCR. We then compared the molecular pattern against the phenotype determined via an in vitro sensitivity test of the clinical isolates against meglumine antimoniate, which is considered the reference technique. Our results demonstrate significant DNA amplification and/or RNA overexpression in 56% of the clinical isolates with the resistant phenotype. All clinical isolates that exhibited significant overexpression of at least 2 genes displayed a resistant phenotype. Among the 14 genes investigated, 10 genes displayed either significant amplification or overexpression in at least 1 clinical isolate; these genes are involved in several metabolic pathways. Moreover, various gene associations were observed depending on the clinical isolates, supporting the multifactorial nature of Leishmania resistance. Molecular resistance features were found in the 3 Leishmania species investigated (Leishmania infantum, Leishmania major, and Leishmania killicki). To our knowledge, this is the first report of the involvement of molecular resistance genes in field isolates of Leishmania major and Leishmania killicki with the resistance phenotype.
在大多数利什曼病流行地区，锑剂仍然是治疗利什曼病各种表现的一线药物，并且已有报道称与寄生虫耐药性相关的治疗失败病例不断增加。在本研究中，我们评估了来自阿尔及利亚、突尼斯和法国南部的47株引起内脏利什曼病和皮肤利什曼病的利什曼原虫临床分离株的分子状态。我们总共检测了14个基因，这些基因在抗锑剂耐药性方面已显示出DNA扩增、mRNA水平或蛋白质表达的显著差异。通过qPCR和qRT-PCR评估每个临床分离株的基因状态。然后，我们将分子模式与通过临床分离株对葡甲胺锑酸盐的体外敏感性试验确定的表型进行比较，葡甲胺锑酸盐体外敏感性试验被视为参考技术。我们的结果表明，56%具有耐药表型的临床分离株存在显著DNA扩增和/或RNA过表达。所有至少有2个基因显著过表达的临床分离株均表现出耐药表型。在所研究的14个基因中，有10个基因在至少1个临床分离株中显示出显著扩增或过表达；这些基因参与了多种代谢途径。此外，根据临床分离株观察到了各种基因关联，这支持了利什曼原虫耐药性的多因素性质。在所研究的3种利什曼原虫（婴儿利什曼原虫、硕大利什曼原虫和基利克利什曼原虫）中均发现了分子耐药特征。据我们所知，这是首次报道分子耐药基因参与具有耐药表型的硕大利什曼原虫和基利克利什曼原虫野外分离株。
