Institut Pasteur, Département de Parasitologie et Mycologie, Immunophysiologie et parasitisme intracellulaire, 25 rue du Dr Roux, 75724 Paris cedex15, France.
Cell Microbiol. 2011 Jan;13(1):81-91. doi: 10.1111/j.1462-5822.2010.01521.x. Epub 2010 Sep 16.
Laboratory mice display features of bona fide hosts for parasites such as Leishmania major and Leishmania donovani. Characterizing the amastigote population size fluctuations and the mouse transcript abundance accounting for these fluctuations demands the capacity to record in real time and integrate quantitative multiparametric datasets from the host tissues where these processes occur. To this end, two technologies, luciferase-expressing Leishmania imaging and a very sensitive quantitative analysis of both Leishmania and mouse transcripts, were combined. After the inoculation of either L. major or L. donovani, the amastigote population size fluctuations - increase, plateau and reduction - were monitored by bioluminescence. It allowed a limited number of mice to be selected for further analysis of both mouse and amastigote transcripts using the real-time quantitative polymerase chain reaction assay we set up. The illustrative examples displayed in the present analysis highlight a correlation between the transcriptional signatures displayed by mouse tissues with the amastigote burden fluctuations. We argue that these two combined technologies will have the potential to provide further insights on complex phenotypes driven by Leishmania developmental programs in the tissues of the mammal hosts.
实验小鼠表现出真正宿主的特征，例如利什曼原虫和杜氏利什曼原虫等寄生虫。要描述无鞭毛体种群大小的波动以及解释这些波动的小鼠转录本丰度，需要能够实时记录并整合来自宿主组织的定量多参数数据集，这些过程发生在宿主组织中。为此，将表达荧光素酶的利什曼虫成像技术与对利什曼原虫和小鼠转录本的非常灵敏的定量分析相结合。在接种利什曼原虫或杜氏利什曼原虫后，通过生物发光监测无鞭毛体种群大小的波动——增加、稳定和减少。这允许选择少数几只小鼠，用于进一步分析我们建立的实时定量聚合酶链反应检测中的小鼠和无鞭毛体转录本。本分析中显示的说明性示例突出了小鼠组织显示的转录特征与无鞭毛体负担波动之间的相关性。我们认为，这两种组合技术将有可能为哺乳动物宿主组织中利什曼虫发育程序驱动的复杂表型提供更深入的见解。
