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从培养物中去除血清会导致婴儿利什曼原虫前鞭毛体生长停滞、倍性改变、感染性降低以及关键基因的差异表达。

Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential Expression of Crucial Genes in Leishmania infantum Promastigotes.

作者信息

Alcolea Pedro J, Alonso Ana, Moreno-Izquierdo Miguel A, Degayón María A, Moreno Inmaculada, Larraga Vicente

机构信息

Department of Molecular Microbiology and Biology of Infections, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CIB-CSIC), Madrid. Spain.

Unit of Microbial Immunology, Centro Nacional de Microbiología, Virología e Inmunología Sanitarias, Instituto de Salud Carlos III (CNM-ISCIII), Majadahonda, Spain.

出版信息

PLoS One. 2016 Mar 9;11(3):e0150172. doi: 10.1371/journal.pone.0150172. eCollection 2016.

Abstract

Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1), the glyoxylase I (GLO1), the hydrophilic surface protein B (HASPB), the methylmalonyl-CoA epimerase (MMCE) and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein.

摘要

婴儿利什曼原虫是引起内脏利什曼病的病原体之一。该物种主要分布在地中海盆地。西班牙最近报告了一次人类疫情。大多数针对利什曼原虫前鞭毛体的操作都采用无细胞培养。该模型稳定且可重复,模拟了沙蝇宿主肠道的条件,而肠道是前鞭毛体发育的自然环境。培养基成分不明确,因为它们含有哺乳动物血清,血清是复杂脂质和蛋白质的丰富来源。血清剥夺会减缓生长动力学,从而降低生物量产量。事实上,我们已经证实生长速率以及感染力都会下降。倍性也会受到影响。关于转录组,高通量方法显示差异表达率较低,但有重要的差异调节基因。最显著的特征是:GINS Psf3、脂肪酰辅酶A合酶(FAS1)、乙二醛酶I(GLO1)、亲水性表面蛋白B(HASPB)、甲基丙二酰辅酶A差向异构酶(MMCE)和一种无鞭毛体蛋白基因上调;磷酸烯醇式丙酮酸羧激酶(gPEPCK)和精氨酸酶下调。本文讨论了其对代谢适应、分化和感染力的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b31/4784933/d1913985f588/pone.0150172.g001.jpg

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