Corrêa José R, Santos Sidney G, Araújo Márcio S, Baptista Cibele, Soares Maurilio J, Brazil Reginaldo P
Lab. Biologia Celular de Microrganismos, Departamento de Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz/FIOCRUZ, Av. Brasil 4365, 21040-900 Rio de Janeiro, RJ, Brazil.
J Parasitol. 2005 Jun;91(3):551-6. doi: 10.1645/GE-396R1.
The present study demonstrates that axenic cultures of Leishmania (Viannia) lainsoni produce larger cell masses in NNN-LIT medium, as well as higher amounts of total proteins in cell extracts, than Leishmania (Leishmania) amazonensis. Antigenicity of L. (V.) lainsoni whole promastigotes is similar to that of L. (L.) amazonensis, as demonstrated by an indirect immunofluorescence diagnostic test using sera from human patients and dogs infected with visceral leishmaniasis. Infectivity of the L. (V.) lainsoni strain used in the present work was demonstrated by the detection by transmission-electron microscopy of tissue amastigotes in skin lesion samples from an experimentally infected hamster. Incubation of lesion fragments in NNN-LIT medium allowed us to obtain promastigote forms, which could be cultivated successfully in vitro. lsoenzyme analysis of such promastigotes confirmed the parasite strain as L. (V.) lainsoni, as compared to other Leishmania reference strains. Our data indicate that L. (V.) lainsoni is a useful alternative source for antigen production as well for use in assays that depend on large cell volumes of Leishmania spp. parasites.
本研究表明，与亚马逊利什曼原虫（Leishmania (Leishmania) amazonensis）相比，纯培养的赖氏利什曼原虫（Leishmania (Viannia) lainsoni）在NNN-LIT培养基中产生的细胞团更大，细胞提取物中的总蛋白含量也更高。使用内脏利什曼病患者和感染犬的血清进行间接免疫荧光诊断试验表明，赖氏利什曼原虫前鞭毛体的抗原性与亚马逊利什曼原虫相似。通过透射电子显微镜检测实验感染仓鼠皮肤病变样本中的组织无鞭毛体，证实了本研究中使用的赖氏利什曼原虫菌株具有感染性。将病变片段在NNN-LIT培养基中培养，使我们能够获得前鞭毛体形式，其可以在体外成功培养。与其他利什曼原虫参考菌株相比，对这些前鞭毛体的同工酶分析证实该寄生虫菌株为赖氏利什曼原虫。我们的数据表明，赖氏利什曼原虫是抗原生产以及用于依赖大量利什曼原虫属寄生虫细胞体积的检测的有用替代来源。
