Cysne-Finkelstein Léa, Silva-Almeida Mariana, Pereira Bernardo Acácio Santini, Dos Santos Charret Karen, Bertho Álvaro Luiz, Bastos Leonardo Soares, de Oliveira Pinto Luzia, de Oliveira Francisco Odêncio Rodrigues, da Souza Pereira Mirian Cláudia, Alves Carlos Roberto
Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Imunoparasitologia - Instituto Oswaldo Cruz - Avenida Brasil, 4365, Manguinhos, 21040-900, Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Biologia Molecular e Doenças Endêmicas - Avenida Brasil, 4365, Manguinhos, 21040-900, Rio de Janeiro, RJ, Brazil.
Protist. 2018 Feb;169(1):107-121. doi: 10.1016/j.protis.2017.11.004. Epub 2017 Dec 5.
The present study demonstrates that the Leishmania (Viannia) braziliensis strain MCAN/BR/1998/R619 is composed of multiple subpopulations with measurable distinctions. Single parasites were separated from a culture of promastigotes in stationary phase by cell sorting and then cultivated as subpopulations. Subsequently, these subpopulations were evaluated for features of in vitro growth, infectivity to murine macrophages and proteinase gene expression. The first evidence of distinct characteristics was observed during the in vitro cultivation of isolated subpopulations, as distinct clusters of patterns were formed among the cultures, indicating the existence of quantifiable fluctuations in metrics. Further, when infecting murine macrophages, the subpopulations induced distinct patterns of production of immune response mediators. While some subpopulations mainly induced the production of IL-1β, IL-6 and TNF-α, others induced the production of IL-12p70 and nitric oxide. Finally, amastigotes of these subpopulations had higher expression of proteinase genes than promastigotes. Additionally, cysteine proteinase, serine proteinase, metalloproteinase and aspartic proteinases were differentially expressed in promastigote and amastigote forms. These data suggest the existence of distinct profiles for the L. (V.) braziliensis MCAN/BR/1998/R619 strain and subpopulations that could drive the success of parasite adaptation to the environments that they inhabit.
本研究表明，巴西利什曼原虫（维安尼亚亚属）菌株MCAN/BR/1998/R619由多个具有可测量差异的亚群组成。通过细胞分选从静止期前鞭毛体培养物中分离出单个寄生虫，然后作为亚群进行培养。随后，对这些亚群的体外生长特征、对小鼠巨噬细胞的感染性和蛋白酶基因表达进行了评估。在分离的亚群体外培养过程中观察到了明显特征的首个证据，因为培养物中形成了不同的模式簇，表明在指标上存在可量化的波动。此外，当感染小鼠巨噬细胞时，亚群诱导了免疫反应介质产生的不同模式。虽然一些亚群主要诱导IL-1β、IL-6和TNF-α的产生，但其他亚群诱导了IL-12p70和一氧化氮的产生。最后，这些亚群的无鞭毛体蛋白酶基因表达高于前鞭毛体。此外，半胱氨酸蛋白酶、丝氨酸蛋白酶、金属蛋白酶和天冬氨酸蛋白酶在前鞭毛体和无鞭毛体形式中差异表达。这些数据表明，巴西利什曼原虫（维安尼亚亚属）MCAN/BR/1998/R619菌株和亚群存在不同的特征，这可能推动寄生虫成功适应它们所栖息的环境。
