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Cell Sorting-Assisted Microarray Profiling of Host Cell Response to Cryptosporidium parvum Infection

机译：细胞分选对宿主细胞对小隐孢子虫感染的反应的微阵列分析

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摘要

To study the transcriptional response of mammalian cells to infection with the intracellular apicomplexan parasite Cryptosporidium parvum, infected and uninfected cells were recovered from C. parvum-infected cell monolayers. This approach, which contrasts with a more conventional experimental design that compares infected to uninfected cell monolayers, enabled the identification of functional categories of genes that are differentially transcribed as a direct consequence of the presence of intracellular parasites. Among several categories of upregulated genes, glycoprotein metabolism was significantly overrepresented. To investigate whether these transcriptional changes affected the composition of the surface of infected cells, cells were probed with fluorescently labeled lectins. Among a panel of seven lectins, soybean agglutinin, which recognizes N-acetyl-d-galactosamine, generated the largest difference in fluorescence between infected and uninfected cells. The origin of the fluorescent signal emitted by infected cells was further investigated and attributed to the overexpression of glycoprotein on the surface of infected cells, as well as the presence of glycoprotein located in the proximity of intracellular parasites.
机译：为了研究哺乳动物细胞对细胞内apicomplexan寄生虫隐孢子虫感染的转录反应，从 C中回收感染和未感染的细胞。感染小细胞的细胞单层。这种方法与比较常规的实验设计（将感染的细胞与未感染的细胞单层进行比较）形成对比，该方法能够鉴定出由于细胞内寄生虫的存在而直接转录的基因的功能类别。在几类上调的基因中，糖蛋白代谢明显过量。为了研究这些转录变化是否影响感染细胞表面的组成，用荧光标记的凝集素探测细胞。在一组七个凝集素中，识别 N -乙酰基-d-半乳糖胺的大豆凝集素在感染细胞和未感染细胞之间产生了最大的荧光差异。进一步研究了被感染细胞发出的荧光信号的起源，并归因于被感染细胞表面糖蛋白的过度表达，以及位于细胞内寄生虫附近的糖蛋白的存在。

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