Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brasil.
Parasit Vectors. 2012 Jan 11;5:11. doi: 10.1186/1756-3305-5-11.
Parasites of the Leishmania genus alternate between the flagellated extracellular promastigote stage and intracellular amastigotes. Here we report the characterization of a Leishmania isolate, obtained from a cutaneous leishmaniasis patient, which presents peculiar morphological features.
The parasite was cultured in vitro and characterized morphologically using optical and electron microscopy. Identification was performed based on monoclonal antibodies and internal ribosomal spacer typing. In vitro macrophage cultures, murine experimental models and sand fly infections were used to evaluate infectivity in vitro and in vivo.
The isolate was identified as Leishmania (Viannia) braziliensis. In the atypical promastigotes grown in culture, a short flagellum surrounded or interrupted by a protuberance of disorganized material was observed. A normal axoneme was present close to the basal body but without elongation much further outside the flagellar pocket. A disorganized swelling at the precocious end of the axoneme coincided with the lack of a paraflagellar rod structure. The isolate was able to infect macrophages in vitro, induce lesions in BALB/c mice and infect Lutzomyia longipalpis.
Notwithstanding the lack of an extracellular flagellum, this isolate infects macrophages in vitro and produces lesions when inoculated into mice. Moreover, it is able to colonize phlebotomine sand flies. Considering the importance attributed to the flagellum in the successful infection and survival of Leishmania in the insect midgut and in the invasion of macrophages, these findings may bring new light into the infectious mechanisms of L. (V.) braziliensis.
利什曼原虫属的寄生虫在鞭毛状的细胞外前鞭毛体阶段和细胞内无鞭毛体之间交替。在这里,我们报告了一种利什曼原虫分离株的特征,该分离株来自皮肤利什曼病患者,具有特殊的形态特征。
寄生虫在体外培养,并通过光学和电子显微镜进行形态学特征描述。鉴定基于单克隆抗体和内部核糖体间隔区分型。体外巨噬细胞培养、小鼠实验模型和沙蝇感染用于评估体外和体内的感染性。
该分离株被鉴定为利什曼原虫(Viannia)巴西利ensis。在培养中生长的非典型前鞭毛体中,观察到短鞭毛周围或被一团紊乱物质的突起所中断。靠近基体存在一个正常的轴丝,但在鞭毛囊外没有进一步伸长。轴丝的早熟端存在一个紊乱的肿胀,与缺乏鞭毛旁棒状结构相对应。该分离株能够感染体外巨噬细胞,在 BALB/c 小鼠中诱导病变,并感染长刺舌蝇。
尽管缺乏细胞外鞭毛,但该分离株能够感染体外巨噬细胞,并在接种小鼠时产生病变。此外,它能够定殖于吸血蝇沙蝇。考虑到鞭毛在利什曼原虫成功感染和在昆虫中肠存活以及入侵巨噬细胞中的重要性,这些发现可能为 L.(V.)巴西利ensis 的感染机制带来新的启示。