Department of Parasitology, Faculty of Medicine, University of Colombo, Sri Lanka.
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Acta Trop. 2023 Sep;245:106979. doi: 10.1016/j.actatropica.2023.106979. Epub 2023 Jun 28.
Leishmaniasis is a neglected tropical disease caused by protozoan parasites of genus Leishmania, and transmitted by different species of Phlebotomine sand flies. More than 20 species of Leishmania are known to cause disease in humans and other animals. Leishmania donovani species complex is known to have a vast diversity of clinical manifestations in humans, but underlying mechanisms for such diversity are yet unknown. Long believed to be strictly asexual, Leishmania have been shown to undergo a cryptic sexual cycle inside its sandfly vector. Natural populations of hybrid parasites have been associated with the rise of atypical clinical outcomes in the Indian subcontinent (ISC). However, formal demonstration of genetic crossing in the major endemic sandfly species in the ISC remain unexplored. Here, we investigated the ability of two distinct variants of L. donovani associated with strikingly different forms of the disease to undergo genetic exchange inside its natural vector, Phlebotomus argentipes. Clinical isolates of L. donovani either from a Sri Lankan cutaneous leishmaniasis (CL) patient or an Indian visceral leishmaniasis (VL) patient were genetically engineered to express different fluorescent proteins and drug-resistance markers and subsequently used as parental strains in experimental sandfly co-infection. After 8 days of infection, sand flies were dissected and midgut promastigotes were transferred into double drug-selective media. Two double drug-resistant, dual fluorescent hybrid cell lines were recovered, which after cloning and whole genome sequencing, were shown to be full genomic hybrids. This study provides the first evidence of L. donovani hybridization within its natural vector Ph. argentipes.
利什曼病是一种被忽视的热带病,由利什曼原虫属的原生动物寄生虫引起,通过不同种类的白蛉沙蝇传播。已知有 20 多种利什曼原虫可导致人类和其他动物患病。利什曼原虫种复合体在人类中已知具有广泛的临床表现多样性,但这种多样性的潜在机制尚不清楚。长期以来,人们一直认为利什曼原虫是严格的无性繁殖,但现已证明它在其白蛉媒介中经历了隐蔽的有性周期。自然种群的杂交寄生虫与印度次大陆(ISC)中不典型临床结果的出现有关。然而,在 ISC 的主要地方性白蛉物种中,遗传杂交的正式证明仍未得到探索。在这里,我们研究了与疾病截然不同形式相关的两种不同变异的利什曼原虫在其自然媒介白蛉(Phlebotomus argentipes)中进行遗传交换的能力。来自斯里兰卡皮肤利什曼病(CL)患者或印度内脏利什曼病(VL)患者的临床分离株的利什曼原虫被遗传工程改造以表达不同的荧光蛋白和药物抗性标记物,随后用作实验性白蛉共感染中的亲本株。感染 8 天后,解剖白蛉并将中肠前鞭毛体转移到双药物选择性培养基中。回收了两种双药抗性、双荧光杂交细胞系,经克隆和全基因组测序后,证实它们是全基因组杂种。这项研究提供了利什曼原虫在其自然媒介白蛉(Ph. argentipes)中发生杂交的第一个证据。
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